bluuebunny/crossref_metadata_2025 huggingface dataset:
Quelques réflexions à propos du 70ème anniversaire de la Convention européenne des droits de l’homme
Authors: Lalumière Catherine
Published: July 2021

À l’occasion de ce soixante-dixième anniversaire, l’auteure propose quelques réflexions sur l’influence de la Convention européenne des droits de l’homme (CEDH) alors que la construction européenne semble menacée. Constituant le socle de cette construction, la CEDH pourrait voir l’importance de son rôle s’accroitre encore, si tant est qu’elle soit capable de faire face aux contestations dont elle est l’objet. Cet article retrace l’histoire, l’impact juridique, la philosophie, mais aussi les tenants politiques du texte européen. Rappelant son ascendance lointaine autant que son esprit humaniste et démocratique construits en réaction aux totalitarismes de l’entre-deux Guerres, l’auteure met en lumière la mutation politique et économique de l’Après-Guerre qui participe à l’affaiblissement de son socle. À l’heure d’une construction politique et économique européenne ébranlée par les volontés souverainistes de certains États, l’auteure invoque les fondements philosophiques de l’idéal européen au secours d’un texte en porte-à-faux, adoubé par les juristes et les humanistes praticiens, méconnu des peuples et craint des gouvernants nationaux. Incarnation de l’Humanisme européen, la CEDH se voit soupçonnée des plus viles turpitudes, elle qui pourtant nous dit tant de la raison d’être de la construction européenne et parait la plus apte à en assurer la survivance et le rayonnement.
La Convention européenne des droits de l’homme et les enjeux bioéthiques
Authors: Bioy Xavier
Published: July 2021

La Convention européenne des droits de l’homme et son interprète authentique n’étaient pas préparés à répondre aux questions sociétales posées par le domaine, en mutation rapide et constante, de la biomédecine. La Convention n’en a pas moins saisi ces questions au prisme des droits subjectifs, de manière souvent tout à fait différente des États eux-mêmes, qui y voyaient plutôt des enjeux de santé publique et d’ordre public, protégeant des valeurs collectives ou laissant au marché le soin de réguler les comportements. La Cour a donc, comme ailleurs, mais avec plus d’audace encore, ouvert son prétoire et élargi le champ des articles 2, 3, 8 et 14 à l’assistance à la reproduction, au prélèvement et usage des éléments corporels, à la recherche biomédicale et à l’accès aux soins innovants. Les techniques biomédicales sont devenues des droits. Ce faisant, la Cour a ressenti la gêne et connu les débats qu’une telle approche suscite entre choix de société, pluralisme et retenue. Il n’est pas anodin de transformer des choix axiologiques nationaux en conflits de droits subjectifs ou de mener à des exceptions via un contrôle concret que les juges nationaux n’utilisent guère. L’usage indécis de la marge d’appréciation en témoigne.
Le Protocole n° 16, entre théories et réalités du dialogue judiciaire
Authors: Burgorgue-Larsen Laurence
Published: July 2021

Cet article a pour ambition de mettre en lumière les différences entre, d’un côté, le texte du Protocole n° 16 – qui présente de nombreuses spécificités théoriques – et, de l’autre, les réalités du dialogue judiciaire qui s’est manifesté à travers les deux saisines de la Cour de cassation française et de la Cour constitutionnelle arménienne. En dépit des intentions louables des auteurs du Protocole n° 16, il n’est pas sûr que ce dernier arrive à engendrer une dynamique dialogique d’envergure, au regard de son caractère facultatif et de l’absence de force obligatoire des avis.
La création du principe de non-refoulement en droit international des droits de la
personne par la Cour européenne des droits de l’homme : un chef-d’oeuvre à protéger
Authors: Delas Olivier
Published: July 2021

Dès les premières années suivant leur création, les organes de contrôle de la
Convention européennes des droits de l’homme vont être saisis d’affaires ayant
trait à des mesures d’éloignement du territoire. Bien que les premiers constats de violation
n’interviennent que bien plus tard, le raisonnement est posé très tôt : lors de la mise en
oeuvre de telles mesures, les États se doivent de ne pas contrevenir à certaines
dispositions de la Convention, notamment son article 3. Par cette construction
jurisprudentielle originale, la Commission et la Cour européennes vont créer un véritable
principe de non-refoulement en droit européen des droits de la personne, qui non seulement
sera repris par les autres juridictions et quasi-juridictions internationales, mais sera
également consacré conventionnellement par les États. Bien plus protecteur que son
prédécesseur en droit international des réfugiés, ce principe n’en est pas moins soumis, en
raison de sa construction même, aux préoccupations souveraines des États. Les enjeux
sécuritaires contemporains, en matière de lutte contre le terrorisme ou d’immigration
irrégulière, font qu’il doit recevoir une attention toute particulière, notamment de la part
de la Cour européenne des droits l’homme elle-même.
The phenomenon of magical treasure hunting as an example for the definition of religious norms in the 18th century
Authors: Michalke Alissa
Published: July 2021

The phenomenon of magical treasure hunting experienced a phase of high popularity in the Holy Roman Empire of German Nation in the early modern period. The applied magical practices were just one part of the wide magical spectrum of that time. In fact, many of them were closely connected to Catholic liturgy. This article will, based on a case study from Bamberg, examine whether or how Catholic doctrine was distinguished from those magical practices and by whom. Furthermore, it will analyse the attitude of clergy and secular people towards magic in order to gain hidden treasure. The study of magical treasure hunting illustrates exemplarily the fight against popular piety and superstition in the XVIIIth century.
Le bios éducatif comme forme de vie normative
Authors: Allouch Hanen
Published: July 2021

Dans cet article, la normativité du milieu éducatif est étudiée à la lumière de représentations littéraires et cinématographiques multiculturelles, italiennes, françaises, québécoises, étasuniennes et maghrébines contemporaines. Un intertexte de classiques met en évidence la persistance et l’évolution de certaines normes dont le calque de la distribution des rôles au sein de la famille patriarcale hétéronomative. Le milieu éducatif normé soumet ses acteurs à l’épreuve de l’espace disciplinaire qui conserve l’idéal du corps sain. Ces fictions critiquent également la normativité coloniale qui imprègne encore les esprits, sur le plan de la représentation et de l’imaginaire.
Immaginare la Grecia oggi, fra stereotipi e contro-narrazioni (street art e flânerie urbana)
Authors: Tentorio Gilda
Published: July 2021

“Greece doesn’t exist” was the provocative title of Michel Grodent’s essay (2000), suggesting the need to overcome all prejudices and stereotypes around the image of Greece. Is this perspective of de-construction possible today? This paper focuses on some specific cultural attempts, following this direction. After a brief overview of the different positions, from enthusiasm to disappointment (idealization, touristic image, the myth of Zorba, financial crisis), it explores two examples of counter-narratives: street art, as an alternative response to hegemonic discourse, and urban flâneries. In particular Christos Chryssòpoulos’ interesting works (Flashlight between Teeth, 2012 and The Flâneur Consciousness, 2015), where text and photography form a dialectic pair and show the present of Athens through its material objects. In both cases, wall-writing and literary photobooks suggest a new gaze – an “apocalyptic” one, according to the etymological root of the word – which reveals throbs, details, microcosms and new perspectives capable of destroying certainties and preconceptions.
Some Results on Cosymplectic Manifolds Admitting Certain Vector Fields
Authors:   Marin Drinov Academic Publishing House, Yoldas Halil
Published: July 2021

The purpose of present paper is to study cosymplectic manifolds admitting certain special vector fields such as holomorphically planar conformal (in short HPC) vector field. First, we prove that an HPC vector field on a cosymplectic manifold is also a Jacobi-type vector field. Then, we obtain the necessary conditions for such a vector field to be Killing. Finally, we give an important characterization for a torse-forming vector field on such a manifold given as to be recurrent.
Structural heterogeneity of cellular K5/K14 filaments as revealed by cryo-electron microscopy
Authors: Weber Miriam S , Eibauer Matthias , Sivagurunathan Suganya , Magin Thomas M , Goldman Robert D , Medalia Ohad
Published: July 2021

Keratin intermediate filaments are an essential and major component of the cytoskeleton in epithelial cells. They form a stable yet dynamic filamentous network extending from the nucleus to the cell periphery, which provides resistance to mechanical stresses. Mutations in keratin genes are related to a variety of epithelial tissue diseases. Despite their importance, the molecular structure of keratin filaments remains largely unknown. In this study, we analyzed the structure of keratin 5/keratin 14 filaments within ghost mouse keratinocytes by cryo-electron microscopy and cryo-electron tomography. By averaging a large number of keratin segments, we have gained insights into the helical architecture of the filaments. Two-dimensional classification revealed profound variations in the diameter of keratin filaments and their subunit organization. Computational reconstitution of filaments of substantial length uncovered a high degree of internal heterogeneity along single filaments, which can contain regions of helical symmetry, regions with less symmetry and regions with significant diameter fluctuations. Cross-section views of filaments revealed that keratins form hollow cylinders consisting of multiple protofilaments, with an electron dense core located in the center of the filament. These findings shed light on the complex and remarkable heterogenic architecture of keratin filaments, suggesting that they are highly flexible, dynamic cytoskeletal structures.
Analysis and tuning of hierarchical topic models based on Renyi entropy approach
Authors: Koltcov Sergei , Ignatenko Vera , Terpilovskii Maxim , Rosso Paolo
Published: July 2021

Hierarchical topic modeling is a potentially powerful instrument for determining topical structures of text collections that additionally allows constructing a hierarchy representing the levels of topic abstractness. However, parameter optimization in hierarchical models, which includes finding an appropriate number of topics at each level of hierarchy, remains a challenging task. In this paper, we propose an approach based on Renyi entropy as a partial solution to the above problem. First, we introduce a Renyi entropy-based metric of quality for hierarchical models. Second, we propose a practical approach to obtaining the “correct” number of topics in hierarchical topic models and show how model hyperparameters should be tuned for that purpose. We test this approach on the datasets with the known number of topics, as determined by the human mark-up, three of these datasets being in the English language and one in Russian. In the numerical experiments, we consider three different hierarchical models: hierarchical latent Dirichlet allocation model (hLDA), hierarchical Pachinko allocation model (hPAM), and hierarchical additive regularization of topic models (hARTM). We demonstrate that the hLDA model possesses a significant level of instability and, moreover, the derived numbers of topics are far from the true numbers for the labeled datasets. For the hPAM model, the Renyi entropy approach allows determining only one level of the data structure. For hARTM model, the proposed approach allows us to estimate the number of topics for two levels of hierarchy.
Population and distribution of wild Asian elephants (Elephas maximus) in Phu Khieo Wildlife Sanctuary, Thailand
Authors: Htet Nyi Nyi Phyo , Chaiyarat Rattanawat , Thongthip Nikorn , Anuracpreeda Panat , Youngpoy Namphung , Chompoopong Phonlugsamee
Published: July 2021

Background

The populations of wild Asian elephants (Elephas maximus) have increased recently after a period of worldwide decline in protected areas. It is important to understand the dynamics and distribution of the remaining populations to ensure their conservation and prevent human-elephant conflicts.

Methods

We monitored the population distribution of elephants between 2016 and 2019 in the Phu Khieo Wildlife Sanctuary, Thailand. We set one hundred forty-nine camera trap locations; cameras recorded 38,834 photos over 6,896 trap nights. Elephants were captured in 4,319 photographs. The maximum entropy modeling software MaxEntwas used to identify elephants’ habitat preferences within 49 of the 149 total camera trap locations according to five environmental factors.

Results

One hundred fourteen elephants were identified. We identified 30 adult males, 43 adult females, 14 sub-adult males, nine sub-adult females, 11 juveniles, and seven calves. The age structure ratio based on adult femaleswas 0.7:1:0.3:0.2:0.3:0.2, and the ratio of reproductive ability between adult females, juveniles, and calves was 1:0.2:0.1. A suitable elephant habitat was determined to be 1,288.9 km2using Area Under the Curve (AUC). An AUC = 0.061 indicated good performance. Our model classified habitat preferences associated with elevation, forests, salt licks, human activity, and slope.

Conclusions

According to our probability map this sanctuary can provide a suitable habitat for elephants. Our results indicate that effective management practices can protect wild Asian elephants in the region and reduce conflict between humans and elephants.
A/r/tografia como processo de percepção de si na constituição enquanto artista-professor-pesquisador em Danças de Salão na contemporaneidade.
Authors: CYPRIANO ABNER SANLAY , CARVALHO CAROLINE
Published: July 2021

À luz de questionamentos de como se constitui um artista-professor-pesquisador, discute-se como os processos de pesquisa a/r/tográfica movem percepções nas dimensões artista-professor-pesquisador em Danças de Salão na contemporaneidade. Um caminhar para si que possibilita, a posteriori, perceber os entrecruzamentos entre essas dimensões bem como pontos de tensionamento que causam processos transicionais entre formas de perceber a experiência da Dança em devir ou após seu ato. Para tal, apresenta-se três obras constituídas durante a graduação em Dança que dialogam com processos de subjetivação do acadêmico. Este opta por uma escrita autobiográfica na tecitura de suas experiências com autores como Fuganti, Bondía, Josso, Dias, Carvalho, entre outros. Também se apresenta o processo de constituição das obras utilizadas na sua monografia de graduação em Dança, em que opta por mergulhar na relação entre marcas e costuras que a experiência deixa em si, revisitando sua infância para que sirva como disparador para essa produção artística. Concluindo-se que este processo de organização de uma pesquisa a/r/tográfica contribui para o reconhecimento de como estes processos se dão de forma imbricada com as experiências vividas.
Dinâmica regional da produção agrícola de Mato Grosso, 2002-2016
Authors: Soares Jadson Gonçalves , Figueiredo Adriano Marcos Rodrigues
Published: July 2021

Na década de 1980 surgiu um novo contexto concorrencial e de oportunidades técnicas, atrelado a uma visão nova e dinâmica do capitalismo em decorrência do processo de internacionalização do capital. Considerando um cenário em que a globalização exige cada vez mais produtividade e especialização das atividades produtivas, em que o produtor e o arranjo produtivo são colocados à prova pelo mercado capitalista, este artigo tem por objetivo verificar a dinâmica regional da produção agrícola de Mato Grosso entre os anos de 2012 e 2016. A revelação dessa dinâmica pode subsidiar a realização de estudos e/ou a formulação de políticas que desenvolvam a estrutura produtiva na esfera estadual. Para tanto realizou-se uma análise regional através de medidas de localização e especialização, identificando as principais transformações no uso do espaço rural. Os resultados apontam para um processo, que ocorre de um ritmo lento a moderado, de mudanças no padrão espacial de localização das principais culturas e de reestruturação produtiva nas regiões. O artigo contribui para o aumento do conhecimento ao estender a pesquisa às áreas geográficas imediatas.
The Rate and Pattern of the Spatio-Temporal Expansion of Rangpur City Corporation, Rangpur, Bangladesh
Authors: Hasnahena . , Sarker Subaran Chandra , Islam Md. Sahidul , Rahman Md. Zakiur
Published: July 2021

Analyzing the spatio-temporal growth of the built-up areas of any urban place is incredibly much vital for the proper planning and development of the urban areas. The present study emphasizes on determining the rate and pattern of spatio-temporal growth of Rangpur City Corporation (RpCC) for the year of 1989, 2000, 2010, and 2017 through Shannon's Entropy with the help of GIS and remote sensing techniques. The Shannon's Entropy technique was adopted in order to determining the dispersion or compactness in the pattern of the built-up areas in the study area. In the present study, Lands at Operational Land Imager (OLI), Lands at Thematic Mapper (TM) and Lands at Enhanced Thematic Mapper plus (ETM+) satellite images on the year of 1989, 2000, 2010, and 2017 were analyzed for certain interpretations. The changes of the built-up areas in RpCC were identified and determined through supervised classification using ArcMap10.5 software. The study indicated that spatio-temporal growth of the built-up areas have been in RpCC was existed during 1989-2017. The built-up areas increased by 5.89 Sq.km. during 1989 -2000, 32.23 Sq.km. during 2000- 2010 and 18.85 Sq.km. during 2010-2017 and the expansion rate of the built-up areas was 8.02%, 25.64% and 6.01% during 1989-2000, 2000-2010 and 2010-2017 respectively. The relative entropy value of 1989, 2000, 2010 and 2017 was found 0.17, 0.24, 0.47 and 0.53 which interprets that the expansion of the built-up areas was existed in RpCC and the pattern of expansion was dispersed. However, the outcomes of this study will be very helpful to formulate perfect planning and management system regarding the expansion of the built-up areas the built-up areas expansion of RpCC.
Adulticidal Activity of Coffee Essential Oils against Two Fruit Fly Species (Tephritidae) under Laboratory Conditions
Authors: Raga Adalton , Silva Sara Braga e , Sousa Ester Marques de , Maia Nilson Borlina
Published: July 2021

The aim of this study was to evaluate the adulticidal effects of green coffee oil (GCO) and roasted coffee oil (RCO) on two species of fruit flies (Tephritidae) include Ceratitis capitata (Wiedemann) and Anastrepha fraterculus (Wiedemann). Adult flies of both species were exposed to GCO and RCO at 2%, 3%, and 4% dilutions in water under Potter spray tower. In general, exposure to GCO and RCO at the three dilutions resulted in higher numbers of dead adults of both species compared to the control populations. The mortalities increased substantially between 60–90 minutes after initial exposure. The 2% GCO dilution reached a maximum of 77.0% corrected mortality of A. fraterculus at 360 minutes. In general, the toxic effect of coffee oils occurred more rapidly in C. capitata than in A. fraterculus. GCO (2% and 3%) and RCO (2%) showed promising results for fruit fly management.
Surveying the Level of Knowledge, Awareness and Health Related Risk of Substance Abuse among Students in University of Port Harcourt’s Host Communities
Authors: Njoku Chigozie , Onyewuchi Nguma Promise
Published: July 2021

Background: Substance abuse refers to the use of all chemicals, drugs and industrial solvents that can produce dependence (psychological and physical). It can also be referred to as the repeated non-medical use of potentially addictive chemical and organic substances. There is poor data about the health related risk of substance abuse among students in Nigeria. Hence, this survey was done to determine the level of knowledge, awareness & the health related risk of substance abuse among students and her host communities as an attempt to curb the menace and its effect on students and the entire society at large. 

Materials and Methods: The study was a descriptive cross-sectional study carried out in ALUU Community in Ikwerre Local Government Area of Rivers State. The study involved 150 volunteers recruited randomly through a multi-staged sampling technique which included secondary school students, undergraduates and postgraduates who are 13yrs and above residing in ALUU community while those who did not give consent were excluded. The data was collected using self-structured close-ended self-administered questionnaires and data analysis done using SPSS version 25. 

Results and Discussions: The results of the study showed that 98.67% have knowledge of drug abuse, the knowledge of health and social effects of substances of abuse on humans was 96.67%. The most prevalent health/social effects were: memory loss 74.67%, sleeping and eating disorders 67.33%. The results of the study have shown that more than 2/3rd of the participants had good knowledge of drug abuse. Similarly, more than 2/3rd of the participants heard about drug/substance abuse via the mass media. The most abused drug/substance was alcohol, 9 in 10 persons abused alcohol. This goes on to imply that the chance that a student who is resident in Aluu abuses alcohol is 90%. 

Conclusion: The results of the study showed that 98.67% have knowledge of drug abuse, the knowledge of health and social effects of substances of abuse on humans was 96.67%. The most prevalent health/social effects were: memory loss 74.67%, sleeping and eating disorders 67.33%. Peer group, neighbourhood influence, and ‘the only child syndrome’ were the attributed contributing factors to the continual use of alcohol/drugs, even when the health and social effects are well known.
Bacteriological Assessment of Automated Teller and Point of Sales Machine User Interface in Uyo Metropolis, Nigeria
Authors: John Ofonime U. M. , Asuquo Christiana E. , Akpan Mfon M.
Published: July 2021

The Automated Teller Machine (ATM) and Point of Sales (POS) machine user interface of different banks, schools, hospitals, restaurants/eateries, shopping malls and petrol/gas stations in Uyo metropolis were bacteriologically assessed in the dry and wet seasons using cultural technique. The result of fomites key pads indicate total heterotrophic bacteria counts (THBC) was in the order: Banks > Hospitals > Schools > Petrol/Gas stations > shopping malls > Restaurants/Eateries in both seasons. There was an increase in the bacterial counts of the fomites from all locations in the wet season compared to the dry season and the difference significant (p = 0.05). The THBC from ATMs in banks, hospitals and schools ranged from 5.6 ± 0.1 to 7.8 ± 0.6 Log₁₀CFU/cm2 and from POS 3.4 ± 0.3 to 5.0 ± 0.2 Log₁₀CFU/cm2) for Petrol/Gas stations, shopping malls and Restaurants/Eateries in the wet season. The dry season revealed THBC range of 4.1± 0.6 to 6.4 ± 0.3 Log₁₀CFU/cm2 for ATMs and 2.6 ± 0.8 to 3.8 ± 0.1 Log₁₀CFU/cm2 for POS. Organisms associated with the fomites were Escherichia coli, Staphylococcus aureus, Staphylococcus epidermidis and species of Bacillus, Pseudomonas, Proteus, Streptococcus, Salmonella, Shigella, Micrococcus, Vibrio and Klebsiella. The highest and least frequency of occurrence in the dry season indicated Micrococcus (17.2%) and Vibrio (1.4%) in relation to Staphylococcus aureus (18.2%) and Proteus sp. (2.8%) in the wet season. The Gram-positive organisms exhibited high and low susceptibility to Gentamycin and Augmentin compared to Ofloxacin and Amoxicillin respectively in Gram-negative bacteria. The results indicate ATM and POS machine user interface as possible sources of pathogenic organisms. Therefore, regular cleaning of ATM and POS user interface and public awareness on the need for adequate personal hygienic practice after the use of these machines is recommended to reduce associated risks.
Erythrocuprein (Cu2Zn2 superoxide dismutase) is the major copper protein of the red blood cell
Authors: Gärtner Alfred , Weser Ulrich
Published: July 2002

The copper balance in the red blood cell deserved special attention as the exact amount of copper being bound in erythrocuprein (Cu2Zn2 superoxide dismutase) is poorly understood. An improved aqueous isolation of erythrocuprein revealed that essentially all erythrocyte copper is found in this protein. This fact is supported by both superoxide dismutase activity measurements of the haemolysate and EPR‐quantification studies throughout the course of the isolation.
Influence of hyperthermia on the phosphorylation of ribosomal protein S6 from human skin fibroblasts and meningioma cells
Authors: Richter W.W. , Zang K.D. , Issinger O.-G.
Published: July 2002

Skin fibroblasts and meningioma cells, derived from primary cultures of the same patients have been used to study the influence of hyperthermia on (i) cell morphology and (ii) phosphorylation pattern of ribosomal and ribosome‐associated proteins. Incubation of tumour cells and fibroblasts up to 7 h at 42°C did not significantly change the cell morphology as compared to control cells kept at 37°C. At 42°C ribosomal protein S6 is shifted cathodically indicating a loss of negative charge, however no quantitative dephosphorylation of S6 was observed. Meningioma cells and fibroblasts did not differ significantly with respect to S6 phosphorylation.
Enhanced polypeptide synthesis programmed by linear DNA fragments in cell‐free extracts lacking exonuclease V
Authors: Jackson Maria , Pratt Julie M. , Holland I.Barry
Published: July 2002

Preparation of an in vitro coupled transcription‐translation system from E. coli strains lacking exonuclease V has greatly improved the system for use with added linear DNA fragments. In fact, in extracts of these mutants linear fragments are stable for several hours. However the cell extracts show a high level of endogenous background. To avoid this complication extracts were prepared at 30°C from a mutant carrying a temperature‐sensitive exonuclease V. Polypeptides coded by a specific DNA region, e.g., delineated by restriction endonuclease sites, can now be easily identified.
Circular dichroism studies on the zinc‐induced conformational changes in S‐100a and S‐100b proteins
Authors: Mani Rajam S. , Kay Cyril M.
Published: July 2002

The effect of Zn2+ binding on the circular dichroism (CD) spectra of brain‐specific S‐100a and S‐100b calcium‐binding proteins has been examined. In the presence of Zn2+, S‐100a undergoes a conformational change and the decrease in ellipticity at 222 nm, as a result of Zn2+ addition, was nearly 1400 deg.cm−2.dmol−1, whereas with S‐100b there was no significant conformational change. Ca2+ was able to bind to S‐100 proteins in the presence of Zn2+ and the two metal‐ion binding sites on the proteins appear to be different. In the presence of Zn2+, K+ had not significant effect on the conformation of S‐100 proteins. Ca2+ and Zn2+ binding induce different environments around the tyrosine residues in S‐100a, whereas with S‐100b, similar changes were observed for the single tyrosine residue, using either metal.
Transfer of ferritin‐bound iron to adriamycin
Authors: Demant Erland J.F.
Published: July 2002

Interactions of adriamycin with ferritin‐bound iron have been investigated. It is demonstrated (i) that adriamycin stimulates an iron‐dependent lipid peroxidation in submitochondrial particles in the presence of ferritin, and (ii) that incubation of adriamycin with ferritin results in a slow transfer of iron to adriamycin with formation of an adriamycin‐iron complex. The results are discussed in relation to the possible role for intracellular iron in adriamycin toxicity.
Increased content of natural ATPase inhibitor in tumor mitochondria
Authors: Luciaková Katarína , Kužela Štefan
Published: July 2002

The ATPase activity of Zajdela hepatoma and Yoshida sarcoma submitochondrial particles was several times lower than the enzyme activity in rat heart and rat liver submitochondrial particles. The content of F1‐ATPase in the tumor mitochondria was found not to be very different from that in mitochondria of rat liver. Immunochemical determination of the amount of the natural ATPase inhibitor revealed that the tumor mitochondria contain 2–3‐times more ATPase inhibitor than control mitochondria. It is concluded that the low ATPase activity of the tumor mitochondria results from the inhibition of the enzyme activity by the natural ATPase inhibitor.
The role of isocitrate in control of the phosphorylation of isocitrate dehydrogenase in Escherichia coli ML308
Authors: El-Mansi E.M.T. , Nimmo H.G. , Holms W.H.
Published: July 2002

Isocitrate dehydrogenase from Escherichia coli is regulated by a reversible phosphorylation mechanism. We confirm here that this permits intracellular isocitrate to rise to a level that can sustain growth on acetate. Isocitrate inhibits isocitrate dehydrogenase kinase and activates isocitrate dehydrogenase phosphatase in vitro. Addition of pyruvate to cultures growing on acetate causes reversible dephosphorylation and activation of isocitrate dehydrogenase, and we show here that this is accompanied by a transient two‐fold increase in the intracellular concentration of isocitrate. The data support our suggestion that isocitrate can play a key role in controlling the phosphorylation state of isocitrate dehydrogenase in vivo.
Monoclonal antibodies against E‐ and F‐type prostaglandins
Authors: Brune K. , Reinke M. , Lanz R. , Peskar B.A.
Published: July 2002

Polyclonal antisera against prostaglandins (PGs) are widely used for the assessment of the biological role of these mediators, but even the most specific contain antibodies against the major metabolites and degradation products of the haptens employed. To overcome this inherent problem we produced monoclonal antibodies (mAs) against PGE2, PGF2α and 6‐keto‐PGF1α using the somatic cell hybridization technique. The mAs against 6‐keto‐PGF1α and PGF2α proved to be highly specific, but allowed only for moderate detection limits (1–2 ng) in conventional fluid phase radioimmunoassays (RIAs). One of the mAs against PGE2 permitted a 100‐fold improvement in the detection limit while being almost devoid of cross‐reactivity with metabolites and other structurally related PGs. These results show that highly specific mAs against PGs can be produced to improve the available RIA technique for PG quantification.
Purification of squalene‐2,3‐epoxide cyclases from cell suspension cultures of Rabdosia japonica Hara
Authors: Abe Ikuro , Ebizuka Yutaka , Seo Shujiro , Sankawa Ushio
Published: July 2002

Microsomes prepared from cell suspension cultures of Rabdosia japonica Hara showed activities for cyclizing squalene‐2,3‐epoxide into cycloartenol, β‐amyrin and α‐amyrin in the presence of Triton X‐100. These activities were efficiently solubilized by treatment with Triton X‐100 and separated by chromatography on hydroxy apatite, DEAE‐cellulose, isoelectric focusing and gel filtration. The purified cycloartenol cyclase showed a single band on SDS‐polyacrylamide gel electrophoresis with Mr = 54000, while β‐amyrin cyclase gave a single band with Mr = 28000.
Maitotoxin, a potent, general activator of phosphoinositide breakdown
Authors: Gusovsky Fabian , Yasumoto Takeshi , Daly John W.
Published: July 2002

Maitotoxin (MTX), a potent marine toxin, elicits a calcium‐dependent activation of cells that can be inhibited by calcium channel blockers like nifedipine. MTX also stimulates phosphoinositide breakdown in smooth muscle cells, NCB‐20 cells and PC12 cells through a nifedipine‐insensitive mechanism. We now report that MTX stimulates phosphoinositide breakdown in a wide variety of cells, and appears to repesent the first general activator of this second messenger‐generating system. MTX‐induced stimulation of phosphoinositide breakdown is dependent in every cell line on the presence of extracellular calcium. In differentiated HL60 cells, in which a chemotactic peptide (fMLP) activates phosphoinositide breakdown via a pertussis toxin‐sensitive mechanism, MTX‐induced stimulation is not affected by pertussis toxin treatment. A phorbol ester has no effect on the response to MTX. Thus, MTX stimulates phosphoinositide breakdown through a calcium‐dependent mechanism that at least in three cell lines (PC12, NCB20 and HL60) is not mediated by a pathway that involves a pertussis toxin‐sensitive guanine nucleotide‐binding protein.
An ATP dependence of mitochondrial F1‐ATPase inactivation by the natural inhibitor protein agrees with the alternating‐site binding‐change mechanism
Authors: Milgrom Ya.M.
Published: July 2002

The rate of inactivation of F1‐ATPase, isolated from beef heart mitochondria, by the active acidic form of the natural inhibitor protein depends on the ATP concentration. An increase in concentration of ATP to ∼ 20 μM leads to a decrease in that of the inhibitor protein inducing 50% inhibition of the F1‐ATPase during 5 s preincubation (C
50); further increase in ATP concentration to 1 mM causes little, if any, change in C
50. However, the C
50 values show a rise at ATP concentrations higher than 1 mM. This ATP dependence of the inhibitor action may be in agreement with a version of the alternating‐site binding‐change mechanism, which assumes that the two‐site catalytic cycle intermediates possessing (i) the products (ADP + Pi) bound in the low‐affinity state at one of the active sites and (ii) an ATP molecule at the other active site are the targets for the acidic form of the inhibitor protein.
Stability‐increasing mutants of glucose dehydrogenase
Authors: Nagao Toshihiro , Makino Yasutaka , Yamamoto Keizo , Urabe Itaru , Okada Hirosuke
Published: July 2002

Four stability‐increasing mutants of glucose dehydrogenase from Bacillus megateriun were purified together with the wild‐type enzyme; each has a single amino acid change of Glu‐96 to Gly, Glu‐96 to Ala, Gln‐252 to Leu, or Tyr‐253 to Cys. These mutant enzymes are more heat resistant at pH 6.5 than the wild‐type enzyme; the replacement of Glu‐96 by Ala increases the thermostability by about 20°C. The mutant enzymes are also resistant to inactivation in alkaline solutions. The replacement of Glu‐96 by Gly or Ala protects the enzyme from alkaline inactivation almost completely. The kinetic constants for the activities of these mutant enzymes do not differ significantly from those of the wild‐type enzyme.
The progenitor of ATP synthases was closely related to the current vacuolar H+‐ATPase
Authors: Nelson Hannah , Nelson Nathan
Published: July 2002

The gene encoding the proteolipid of the vacuolar H+‐ATPase of yeast was cloned and sequenced. The deduced amino acid sequence of the yeast protein is highly homologous to that of the proteolipid from bovine chromaffin granules. In contrast to other membrane proteins the transmembrane segments of the bovine and yeast proteolipids were much more conserved than the hydrophilic parts. The fourth transmembrane segment, which contains the DCCD‐binding site, was conserved 100%. Comparison of vacuolar and eubacterial proteolipids revealed a homology which pointed to a common ancestral gene that underwent gene duplication to form the vacuolar proteolipids. Additional support for this notion came from the amino acid sequences of subunits involved in the catalytic sectors of archaebacterial ATP synthase and plant and yeast vacuolar H+‐ATPases, which reveal extensive sequence homology. Slight, but significant, homology between the archaebacterial and eubacterial ATP synthases was observed. These observations might suggest that the progenitor of ATP synthases was closely related to the present vacuolar H+‐ATPases.
The amino acid sequence of a novel inhibitor of cathepsin D from potato
Authors: Ritonja Anka , Križaj Igor , Meško Pika , Kopitar Majda , Lučovnik Peter , Štrukelj Borut , Pungerčar Jože , Buttle David J. , Barrett Alan J. , Turk Vito
Published: July 2002

The amino acid sequence of a cathepsin D inhibitor isolated from potato is described. It was determined by analysis of peptides generated by use of the glycine‐specific proteinase PPIV. The order of the peptides was established by examination of tryptic peptides derived from the two cyanogen bromide peptides. The inhibitor comprises 187 amino acid residues, and has a calculated M
r of 20 450.
Inactivation of the uterine estrogen receptor binding of estradiol during P‐450 catalyzed metabolism of chlorotrianisene (TACE)
Authors: Kupfer David , Bulger William H.
Published: July 2002

Chlorotrianisene (TACE) exhibits in vitro little or no binding to the uterine estrogen receptor (ER) but demonstrates potent estrogenic activity in vivo, indicating that TACE is a proestrogen/proantiestrogen. Our earlier studies demonstrated that the incubation of TACE with rat liver microsomes and NADPH generates a reactive intermediate (T∗) which binds covalently to proteins. The current study examined the possibility that T∗ may inactivate the uterine ER. The incubation of TACE with rat liver microsomes and NADPH in the presence of rat uteri, under conditions which generate T∗, markedly decreased the binding capacity of the ER for [3H]estradiol (E2). The evidence indicates that ER inactivation was probably due to irreversible (covalent) binding of T∗ to the E2 binding site. The possibility that the antiestrogenic action of TACE and of other triphenylethylenes involves such a novel mechanism is discussed.
Tetanus toxin‐induced protein kinase C activation and elevated serotonin levels in the perinatal rat brain
Authors: Aguilera Jose , Lopez Lluis A. , Yavin Ephraim
Published: July 2002

A single intracerebral injection of tetanus toxin (TeTox) is able to produce a time‐dependent translocation of Ca2+‐phosphatidylserine‐dependent protein kinase C (PKC) in close‐to‐tenn rat brain. TeTox‐triggered translocation of PKC is dose‐ and time‐dependent, can be prevented by tetanus antitoxin, and does not occur upon administration of toxin fragments B and C. TeTox‐triggered PKC translocation is accompanied by a timedependent increase in brain serotonin (5‐HT). Increase of brain 5‐HT is independent of monoamine oxidase inhibition by pargyline. Phorbol ester and TeTox cause a significant increase in serotonin while H‐7, a kinase inhibitor, does not affect serotonin levels but abolishes the effect of TeTox. Gangliosides prevent TeTox‐triggered 5‐HT increase. The data are consistent with the possibility that TeTox acts effectively on the serotonergic innervation, presumably in conjunction with PKC to cause accumulation of serotonin.
Purification and study of a bacterial glutathione S‐transferase
Authors: Arca Pilar , García Pilar , Hardisson Carlos , Suárez Juan E.
Published: July 2002

A glutathione S‐transferase from Escherichia coli has been purified approximately 800‐fold with an 11% activity yield by passage through DEAE Sephacel and glutathione‐agarose affinity columns. Its functional form is a homodimer of two 24 000 Da polypeptides that catalyzes the binding of glutathione and 1‐chloro‐2,4‐dinitrobenzene withK
m values of 0.25 and 1.5 mM, respectively. Optima of pH and temperature were 7.5 and 35°C. The activity was stimulated (30%) by ethylenediaminetetraacetic add. The N‐tenninal amino acid sequence was: Met‐Leu‐Leu‐Phe‐Ile‐Leu‐Pro‐Gly‐Ala.
Precursor proteins in transit through mitochondrial contact sites interact with hsp70 in the matrix
Authors: Ostermann Joachim , Voos Wolfgang , Kang Pil Jung , Craig Elizabeth A. , Neupert Walter , Pfanner Nikolaus
Published: July 2002

We previously reported that hsp70 in the mitochondrial matrix (mt‐hsp70 = Ssc1p) is required for import of precursor proteins destined for the matrix or intermembrane space. Here we show that mt‐hsp70 is also needed for the import of mitochondrial inner membrane proteins. In particular, the precursor of ADP/ATP carrier that is known not to interact with hsp60 on its assembly pathway requires functional mt‐hsp70 for import, suggesting a general role of mt‐hsp70 in membrane translocation of precursors. Moreover, a precursor arrested in contact sites was specifically co‐precipitated with antibodies directed against mt‐hsp70. We conclude that mt‐hsp70 is directly involved in the translocation of many, if not all, precursor proteins that are transported across the inner membrane.
The three‐dimensional structure of proteasomes from Thermoplasma acidophilum as determined by electron microscopy using random conical tilting
Authors: Hegerl Reiner , Pfeifer Günter , Pühler Gabriela , Dahlmann Burkhardt , Baumeister Wolfgang
Published: July 2002

The three‐dimensional structure of proteasomes from the archaebacterium Thermoplasma acidophilum has been determined to a resolution of approximately 2 nm from electron micrographs of negatively stained preparations using the method of random conical tilting. The particles turn out to be essentially cylinder‐shaped barrels, 15 nm long and 11 nm wide, enclosing a tripartite inner compartiment. An account is given of some of the present limitations which prevent to attain a higher resolution and possible ways to overcome these limitations are indicated.
A plant homologue to mammalian brain 14‐3‐3 protein and protein kinase C inhibitor
Authors: Hirsch Stephan , Aitken Alastair , Bertsch Uwe , Soll Jürgen
Published: July 2002

We have isolated cDNA clones of Spinacea oleracea L. and Oenothera hookeri of 930 and 1017 base pairs, respectively. The open reading frame deduced from the Oenothera sequence codes for a protein of a calculated molecular mass of 29 200. The primary amino acid sequence exhibits a very high degree (88%) of homology to the 14‐3‐3 protein from bovine brain, and protein kinase C inhibitor from sheep brain. Subsequently the plant protein was partially purified from leaf extract. The partially purified plant protein inhibited protein kinase C from sheep brain in a heterologous assay system. The active fraction consisted of 5–6 different polypeptides of similar molecular size. One of these proteins crossreacted with a peptide‐specific antibody against protein kinase C inhibitor protein from sheep brain.
Cyclosporin A—cyclophilin complex formation A model based on X‐ray and NMR data
Authors: Spitzfaden Claus , Weber Hans-Peter , Braun Werner , Kallen Joerg , Wider Gerhard , Widmer Hans , Walkinshaw Malcolm D. , Wüthrich Kurt
Published: July 2002

The previously determined 3D NMR solution structure of cyclophilin‐bound cyclosporin A (CsA) was docked onto the X‐ray crystal structure of cyclophilin. Intermolecular nuclear Overhauser effects (NOE) between CsA and cyclophilin were used as constraints in a restrained energy minimization to generate a model of the complex which satisfied all the NOE distance constraints. The model shows that the residues 9 to 11 and 1 to 5 of the cyclic CsA molecule are in contact with cyclophilin. Comparing the model of the CsA—cyclophilin complex to the X‐ray crystal structure of a complex of cyclophilin with a substrate for peptidyl‐proline cis‐trans isomerase activity, i.e. the linear tetrapeptide substrate ae‐Ala‐Ala‐Pro‐Ala‐amc (ac. acetyl; amc. amidomethylcoumarin), one notices that the contacting peptide segments in the two ligands are oriented in opposite directions, and that the side chain or MeVal‐11 of CsA superposes rather precisely with the position of the prolyl residue in ae‐Ala‐Ala‐Pro‐Ala‐amc.
Cytoplasmic triacylglycerols and cholesteryl esters are degraded in two separate catabolic pools in cultured human fibroblasts
Authors: Hilaire Nathalie , Nègre-Salvayre Anne , Salvayre Robert
Published: July 2002

The sources and the catabolic pathways of cytoplasmic pools of triacylglycerols and cholesteryl esters have been comparatively investigated in cultured fibroblasts from normal subjects and from patients affected with neutral lipid storage disease (NLSD) and Wolman disease (WD). (i) Endogenously biosynthesized triacylglycerols and cholesteryl esters were degraded extra‐lysosomaily since they were catabolized at similar rates in normal and in WD fibroblasts. In NLSD fibroblasts, the degradation of endogenous triacylglycerols was severely deficient, whereas that of endogenous cholesteryl esters was in the normal range, (ii) Reconstituted high density lipoproteins (HDL) containing radiolabelled [3H]triolein and cholesteryl [14C]oleate were taken up by cultured fibroblasts and rapidly degraded in a non‐lysosomal compartment. In NLSD fibroblasts the degradation of HDL‐[3H]triolein was blocked whereas that of HDL‐[14C]cholesteryl oleate was in the normal range. These data suggest that: (i) the cytoplasmic pools of triacylglycerols and cholesteryl esters originate from HDL uptake and from endogenous biosynthesis as well; (ii) cytoplasmic (non‐lysosomal) triacylglycerols and cholesteryl esters are degraded by two separate catabolic pathways.
A metal‐linked gapped zipper model is proposed for theh sp90‐glucocorticoid receptor interaction
Authors: Schwartz J.A. , Mizukami H. , Skafar D.F.
Published: July 2002

In the presence of certain metals, regions of the hormone binding domain of the glucocorticoid receptor (GR) are capable of binding the 90 kDa heat shock protein (hsp90). Using secondary structure prediction methods in correlation with the experimental data, we propose a model which predicts the presence of two widely spaced leucine zipper‐like heptads on either side of a central subdomain. The heptads could interact hydrophobically with similar regions on the hsp90 homodimer, bringing putative metal binding residues on each protein close enough to establish a shared metal bridge. The central subdomain between heptads is suggested to contain regions involved in metal binding, steroid binding, and conformational mobility. The hypothetical model that we are proposing therefore addresses the nature of the structural link between hsp90 binding, hormone binding, and conformational changes in the receptor.
Functional analysis of block 5, one of the highly conserved amino acid sequences in the 130‐kDa CryIVA protein produced by Bacillus thuringiensis subsp. israelensis
Authors: Nishimoto Tomoyuki , Yoshisue Hajime , Ihara Kentaro , Sakai Hiroshi , Komano Tohru
Published: July 2002

There are five amino acid sequences highly conserved among Bacillus thuringiensis δ‐endotoxins. We have changed the amino acid residues in block 5, one of the conserved sequences, of CryIVA. When the amino acid residues with charged side chains were replaced by others, the amount of production of the altered CryIVA protein was markedly decreased. It is suggested that the decrease is caused by the unstable conformation of the altered CryIVA protein molecule, as judged by digestion with trypsin and thermolysin. On the other hand, the substitution of amino acid residues in block 5 did not affect the insecticidal activity of CryIVA. These results strongly suggest that block 5 of CryIVA is one of the stability‐determining elements of the protoxin molecule.
X‐Ray crystallographic studies of recombinant inorganic pyrophosphatase from Escherichia coli
Authors: Oganessyan V.Yu. , Kurilova S.A. , Vorobyeva N.N. , Nazarova T.I. , Popov A.N. , Lebedev A.A. , Avaeva S.M. , Harutyunyan E.H.
Published: July 2002

An E. coli inorganic pyrophosphatase overproducer and a method for a large‐scale production of the homogeneous enzyme are described. The inorganic pyrophosphatase was crystallized in the form containing one subunit of a homohexameric molecule per asymmetric unit: space group R32, a = 110.4 Å, c = 76.8 Å. The electron density map to 2.5 Å resolution phased with Eu‐ and Hg‐derivatives (figure of merit,  = 0.51) was improved by the solvent flattening procedure ( = 0.77). The course of the polypeptide chain and the secondary structure elements, intersubunit contacts and positions of the active sites were characterized. Homology with S. cerevisiae inorganic pyrophosphatase structure was found.