bluuebunny/crossref_metadata_2025 huggingface dataset:
Fostering Employability through Versatility within Specialisation in Medical Translation Education
Authors: Muñoz-Miquel Ana , Montalt Vicent , García-Izquierdo Isabel
Published: July 2020

The current increase of automation (Rodríguez 2017; Massey/Wieder 2018) and the emergence of new needs and forms of communication are triggering substantial changes in the translation profession, in the role of the translator and in translator education. Previous studies (Muñoz-Miquel 2014, 2016a, 2018) have shown the rich variety of tasks — beyond those traditionally considered — that medical translators perform in the workplace, including heterofunctional translation, editing, or community management. The ability to develop new skills and to adapt continuously to the changing needs of the market is one of the essential characteristics of the translator in the 21st century. That is why we consider it of critical importance that, together with specialisation, versatility is promoted in translator education. In this article, we explore the notion of versatility and propose a teaching strategy that incorporates it and can contribute to improving the employability of future translators. Specifically, we put forward some pedagogical proposals for the English-Spanish language combination that promote diversification of competences and tasks within a narrow specialisation — the medical and healthcare field — in order to provide (future) translators with the versatility necessary to respond to new demands and thus be more employable. Our approach is based on the results of surveys of professional medical translators on the tasks and roles they perform, as well as on our own teaching experience in a master’s degree programme in medical translation.
Verdenshavet og Frederiksholms Kanal: Viden er magt
Authors: Udenrigs Redaktion
Published: July 2020

Intet resumé
Den tikkende miljøbombe i øst
Authors: Hansen Morten
Published: July 2020

Intet resumé
Killer Folklore
Authors: Barnes Naomie
Published: July 2020

This article considers the development of identity within the True Crime Community (TCC) and examines the contagion theory of crime surrounding serial and mass murder within the United States. The TCC is an online Tumblr group that shares pictures, jokes, legends and other narratives surrounding such killers. TCC members are frequently blamed by outsiders for creating spaces where killers become celebrities which, in theory, creates more killers. Two issues arise out of contagion theory. The first is that modes of participation with crime representations and the production of texts around true crimes is shaped by contagion theory and vernacular debates about fandom. Second, contagion theory itself is open to critical cross disciplinary investigation for which folklorists, examining communities who participate in (re)producing true crime texts can offer valuable data, approaches and theory. Moreover, as folklorists working in rumour and contemporary legend have noted, we may have a role to play in interrupting crime cycles themselves.
The Problem with Changing the Folk Law of Bullying
Authors: Learning Jeffery
Published: July 2020

In this paper I will explore the crucial relationship between folk law and bullying in an effort to explain some of the most problematic pieces being left out of current anti-bullying solutions in Newfoundland and Labrador. Using bullying examples from interviews with teachers from across the province of Newfoundland and Labrador collected for my dissertation, “Dialogues of Dominance: Narrative, Occupational Folklore, & the Bullying of Public-School Teachers”, I will attempt to show how insidious the bullying problem really is. I will focus on three key barriers facing school anti-bullying movements in Canada today: the exploitation of folk law; the definitional confusion of ‘bullying’; and the creation and maintenance of a triviality barrier that obfuscates bullying when it happens between adults. I will then use these problems to explain why it falls to the use of folklore to curb this behaviour, and how folkloristics can help with the development of stronger anti-bullying campaigns.
The Speaking Body and the Silent Mouth
Authors: Bodner John
Published: July 2020

This paper analyzes two micro-case studies to expand our understanding of communication techniques in criminal practices. In the first instance the torture of a suspected drug thief by two street kid research participants is investigated as both an instrumental trial by ordeal and a communication strategy in a media poor community. In the second case study, excerpts from occupational folklore and lifestory interviews are presented to demonstrate the complex ways in which silence is used and resisted in the stories of illegal marijuana growers in a small village in British Columbia.
The phenomenon of personality of T. L. Schepkina-Kupernik in the context of theatrical process of the late XIX – early XX century
Authors: Liubivaia Irina , Korol'kova Natal'ya Evgen'evna
Published: July 2020

The interest to the almost forgotten names of female writers of the Silver Ages gas grown over the recent decades. Among them is a prominent and talented Russian and Soviet poetess, translator, writer, play writer, memoirist, journalist T. L. Schepkina-Kupernik. She dedicated approximately 60 years of her life to literary work, and in 1940 was awarded the title of “Honored Master of Arts of RSFSR”. Relevance of the article is defined by the fact that presently the artistic heritage of T. L. Schepkina-Kupernik became undeservingly forgotten, even though it represents great value not only for the Russia, but also for word art history, especially for historians of theatre and literature. Thanks to the translations of T. L. Schepkina-Kupernik, the Russian stage enriched its repertoire, and the audience became acquainted with world-renown dramaturgists. Connoisseurs of the theatrical art were able to appreciate performance of the actors thanks to the memoirs on the actors and peer reviews on their stage performances. An important detail in biography of Tatiana Lvovna consists in her personal experience of acting on stage. The author analyzes the persona of Schepkina-Kupernik from various aspects of her creative work. Analysis is conducted on her work as an actress, writer, translator, and memoirist. Her contribution to the history of the Russian theater is reviewed.
Different dendritic domains of the GnRH neuron underlie the pulse and surge modes of GnRH secretion in female mice
Authors: Wang Li , Guo Wenya , Shen Xi , Yeo Shel , Long Hui , Wang Zhexuan , Lyu Qifeng , Herbison Allan E , Kuang Yanping
Published: July 2020

The gonadotropin-releasing hormone (GnRH) neurons exhibit pulse and surge modes of activity to control fertility. They also exhibit an unusual bipolar morphology comprised of a classical soma-proximal dendritic zone and an elongated secretory process that can operate as both a dendrite and an axon, termed a ‘dendron’. We show using expansion microscopy that the highest density of synaptic inputs to a GnRH neuron exists at its distal dendron. In vivo, selective chemogenetic inhibition of the GnRH neuron distal dendron abolishes the luteinizing hormone (LH) surge and markedly dampens LH pulses. In contrast, inhibitory chemogenetic and optogenetic strategies targeting the GnRH neuron soma-proximal dendritic zone abolish the LH surge but have no effect upon LH pulsatility. These observations indicate that electrical activity at the soma-proximal dendrites of the GnRH neuron is only essential for the LH surge while the distal dendron represents an autonomous zone where synaptic integration drives pulsatile GnRH secretion.
Assigning mitochondrial localization of dual localized proteins using a yeast Bi-Genomic Mitochondrial-Split-GFP
Authors: Bader Gaétan , Enkler Ludovic , Araiso Yuhei , Hemmerle Marine , Binko Krystyna , Baranowska Emilia , De Craene Johan-Owen , Ruer-Laventie Julie , Pieters Jean , Tribouillard-Tanvier Déborah , Senger Bruno , di Rago Jean-Paul , Friant Sylvie , Kucharczyk Roza , Becker Hubert Dominique
Published: July 2020

A single nuclear gene can be translated into a dual localized protein that distributes between the cytosol and mitochondria. Accumulating evidences show that mitoproteomes contain lots of these dual localized proteins termed echoforms. Unraveling the existence of mitochondrial echoforms using current GFP (Green Fluorescent Protein) fusion microscopy approaches is extremely difficult because the GFP signal of the cytosolic echoform will almost inevitably mask that of the mitochondrial echoform. We therefore engineered a yeast strain expressing a new type of Split-GFP that we termed Bi-Genomic Mitochondrial-Split-GFP (BiG Mito-Split-GFP). Because one moiety of the GFP is translated from the mitochondrial machinery while the other is fused to the nuclear-encoded protein of interest translated in the cytosol, the self-reassembly of this Bi-Genomic-encoded Split-GFP is confined to mitochondria. We could authenticate the mitochondrial importability of any protein or echoform from yeast, but also from other organisms such as the human Argonaute 2 mitochondrial echoform.
Infant and adult SCA13 mutations differentially affect Purkinje cell excitability, maturation, and viability in vivo
Authors: Hsieh Jui-Yi , Ulrich Brittany N , Issa Fadi A , Lin Meng-chin A , Brown Brandon , Papazian Diane M
Published: July 2020

Mutations in KCNC3, which encodes the Kv3.3 K+ channel, cause spinocerebellar ataxia 13 (SCA13). SCA13 exists in distinct forms with onset in infancy or adulthood. Using zebrafish, we tested the hypothesis that infant- and adult-onset mutations differentially affect the excitability and viability of Purkinje cells in vivo during cerebellar development. An infant-onset mutation dramatically and transiently increased Purkinje cell excitability, stunted process extension, impaired dendritic branching and synaptogenesis, and caused rapid cell death during cerebellar development. Reducing excitability increased early Purkinje cell survival. In contrast, an adult-onset mutation did not significantly alter basal tonic firing in Purkinje cells, but reduced excitability during evoked high frequency spiking. Purkinje cells expressing the adult-onset mutation matured normally and did not degenerate during cerebellar development. Our results suggest that differential changes in the excitability of cerebellar neurons contribute to the distinct ages of onset and timing of cerebellar degeneration in infant- and adult-onset SCA13.
Molecular phylogenetics and evolutionary history of the endemic land snail genusEverettiain northern Borneo
Authors: Liew Thor-Seng , Marzuki Mohammad Effendi , Schilthuizen Menno , Chen Yansen , Vermeulen Jaap J. , Mohd-Azlan Jayasilan
Published: July 2020

Borneo has gone through dramatic changes in geology and topography from the early Eocene until the early Pliocene and experienced climatic cycling during the Pleistocene. However, how these changes have shaped the present-day patterns of high diversity and complex distribution are still poorly understood. In this study, we use integrative approaches by estimating phylogenetic relationships, divergence time, and current and past niche suitability for the Bornean endemic land snail genusEverettiato provide additional insight into the evolutionary history of this genus in northern Borneo in the light of the geological vicariance events and climatic fluctuations in the Pleistocene. Our results show that northern BorneoEverettiaspecies belong to two deeply divergent lineages: one contains the species that inhabit high elevation at the central mountain range, while the other contains lowland species. Species diversification in these lineages has taken place before the Pliocene. Climate changes during the Pleistocene did not play a significant role in species diversification but could have shaped contemporary species distribution patterns. Our results also show that the species-rich highland habitats have acted as interglacial refugia for highland species. This study of a relatively sedentary invertebrate supports and enhances the growing understanding of the evolutionary history of Borneo. Species diversification inEverettiais caused by geological vicariance events between the early Miocene and the Pliocene, and the distribution patterns were subsequently determined by climatic fluctuations in the Pleistocene.
Ejemplar completo
Authors: Derecho y Ciencia Política Revista
Published: July 2020

Volumen 10, número1.
Perception of nursing workers about the workloads in a neonatal intensive care unit
Authors: Michaello Renata da Silva , Tomaschewski-Barlem Jamila Geri , Carvalho Deciane Pintanela de , Rocha Laurelize Pereira , Bordignon Simoní Saraiva , Neutzling Bruna Ruoso da Silva
Published: July 2020

Objetivo: Conhecer a percepção dos trabalhadores de enfermagem acerca das cargas de trabalho em uma Unidade de Terapia Intensiva Neonatal. Método: Estudo qualitativo com 24 trabalhadores de enfermagem em uma unidade de Terapia Intensiva Neonatal em um hospital universitário, em junho de 2017, por meio de entrevistas semiestruturadas e analisadas por análise textual discursiva. Resultados: Verificou-se que os trabalhadores de enfermagem percebem sua exposição às cargas de trabalho e as relacionam com a ocorrência de acidentes e doenças de trabalho. As cargas psíquicas foram as mais representativas no ambiente de trabalho, entretanto, destaca-se a relação identificada pelos trabalhadores entre as cargas químicas e os produtos utilizados na limpeza das incubadoras. Conclusão: O reconhecimento das cargas de trabalho oportuniza a criação de medidas que possam minimizá-las durante a assistência de recém-nascidos em cuidado intensivo, visando à saúde do trabalhador.
Sexualidade de mulheres submetidas à mastectomia: identificação das fases afetadas no ciclo da resposta sexual
Authors: Martins Juliana Oliveira de Araújo , Santos Amuzza Aylla Pereira dos , Holanda Juliana Bento de Lima , Trindade Ruth França Cizino da , Lima Luana Karla Pessoa
Published: July 2020

Objetivo deste estudo foi analisar a sexualidade das mulheres mastectomizadas, identificando quais as fases do ciclo da resposta sexual são afetadas e quais as disfunções desenvolvidas, utilizando um questionário com dados socioeconômicos e o Questionário Quociente Sexual - versão feminina. Trata-se de uma pesquisa descritiva de cunho quantitativo, com mulheres mastectomizadas em atendimento no ambulatório de Cuidado oncológicos. O desempenho sexual da população estudada apresentou-se heterogêneo, onde 50% apresentou padrão sexual satisfatório e 50% algum tipo de comprometimento na sexualidade, e dentro deste grupo, incluem-se as casadas, com menos de 49 anos e com grau de instrução de nível fundamental. Diante do exposto, os profissionais de saúde em geral devem estar atentos para a ocorrência dessas modificações na vida dessa mulher, diante do todo o processo, desde diagnóstico, no tratamento e readaptação, para dessa forma contribuir para o cuidado integral.
Deoxyribose breakdown by the adriamycin semiquinone and H2O2: evidence for hydroxyl radical participation
Authors: Bates Diana A. , Winterbourn Christine C.
Published: July 2002

We report our finding that the reaction between the adriamycin semiquinone (produced by reduction of the drug by xanthine oxidase) and H2O2 in N2 causes deoxyribose degradation to a thiobarbituric acid‐reactive chromogen. Deoxyribose breakdown was inhibited by scavengers of hydroxyl radicals, providing evidence for the participation of hydroxyl radicals. The reaction was detected in air, but was less efficient in air than in N2. Deoxyribose degradation did not require a metal catalyst, and was inhibited by superoxide dismutase in air, but not N2. A similar reaction with deoxyribose in DNA may be of major importance in the antitumour action of adriamycin.
The purification and partial amino acid sequence of a polypeptide from the glutelin fraction of rice grains; homology to pea legumin
Authors: Zhao Wen-Ming , Gatehouse John A. , Boulter Donald
Published: July 2002

The glutelin fraction was extracted from grain meals of rice (Oryzea sativa) with 50 mM Tris‐HCl buffer (pH 8.8) containing 6 M urea and 10 mM 2‐mercaptoethanol. Polypeptides of glutelin were separated and purified by ion‐exchange chromatography under denaturing conditions. Analysis by two‐dimensional gel electrophoresis showed that 2 major polypeptides of the rice glutelin fraction, M
r 36 000 and 22 000, were linked in disulphide bonded pairs containing one M
r 36 000 and one M
r 22 000 subunit. A partial amino acid sequence of the purified M
r 22 000 glutelin subunit showed it to be homologous to the β‐subunit of pea legumin, a storage protein which also contains disulphide‐linked subunit pairs (M
r 38 000 and M
r 22 000). It is therefore proposed that the major component of rice glutelin is a legumin‐like protein.
The complete sequence of human preprocalcitonin
Authors: Le Moullec J.M. , Jullienne A. , Chenais J. , Lasmoles F. , Guliana J.M. , Milhaud G. , Moukhtar M.S.
Published: July 2002

DNA complementary to mRNA extracted from the thyroid glands of patients suffering from medullary carcinoma of the thyroid (MCT), a calcitonin‐producing tumour, was inserted in the Pst site of pBR 322 by G‐C tailing. The recombinant plasmids were used to transform Escherichia coli DP 50. Ampicillin‐resistant clones were screened using a 32P‐labelled cDNA to mRNA extracted from a case of MCT particularly rich in calcitonin (CT) mRNA. Positive clones were subsequently rescreened using a 32P poly(T) probe. Eighty clones were thus purified, and the inserts obtained by digestion with PstI were subjected to positive hybridization selection with subsequent translation in vitro. An insert stimulating synthesis of the protein and containing restriction sites compatible with the previously published complete sequence of calcitonin mRNA from rat was sequenced. This cDNA insert contained the entire coding region of 426 bp, 70 bp at the 5′‐end, and 295 bp upstream from the poly(A) tail. The complete amino acid sequence of human preprocalcitonin could thus be deduced.
Chemically modified low density lipoproteins as inducers of enzyme release from macrophages
Authors: Hartung Hans-Peter , Kladetzky Rudolf G. , Hennerici Michael
Published: July 2002

Macrophages carry receptors on their surface for acetylated low density lipoprotein (ac‐LDL). Receptor‐mediated endocytosis of ac‐LDL is followed by intracellular cholesterol accumulation. We investigated whether occupation of these binding sites evokes the release of hydrolytic enzymes from mouse peritoneal macrophages cultured for up to 48 h. ac‐LDL at concentrations ranging from 25–250 was noted to promote in a dose‐dependent fashion secretion of the neutral proteinase elastase (EC 3.4.21.37) and the lysosomal acid hydrolases N‐acetyl‐β‐glucosaminidase (EC 3.2.1.30), β‐glucuronidase (EC 3.2.1.31), β‐galactosidase (EC 3.2.1.23), α‐mannosidase (EC 3.2.1.24) and cathepsin D (EC 3.4.23.5). This stimulatory effect was non‐cytotoxic. LDL modified by treatment with malondialdehyde was also capable of augmenting enzyme liberation into culture supernates. These findings may have implications for some aspects of the atherosclerotic process.
Electrogenic reduction of Rhodospirillum rubrum reaction centre bacteriochlorophyll P870+ by redox dyes
Authors: Drachev Lel A. , Kaminskaya Olga P. , Konstantinov Alexander A. , Semenov Alexey Yu. , Skulachev Vladimir P.
Published: July 2002

Rhodospirillum rubrum
Membrane potential
Proteoliposome
Reaction center bacteriochlorophyll
Intramolecular electron transfer
Rapid kinetics
Biochemical comparison of γ‐crystallins from duck and frog eye lenses
Authors: Chiou Shyh-Horng , Chang Wen-Chang , Kuo Jane , Pan Fu-Ming , Lo Tung-Bin
Published: July 2002

A biochemical comparison has been made on the crystallins isolated from duck and frog lenses. Gel‐permeation chromatography of lens homogenates from both classes on Fractogel TSK HW‐55(S) revealed a homogeneous trimeric protein of 120 kDa in the duck lenses and a monomeric protein of 39 kDa in the frog lenses. Both crystallin fractions consist only of an approx. 38‐kDa polypeptide in their subunit structures as determined by SDS gel electrophoresis. These two crystalline were compared with respect to their native molecular masses, subunit structures, peptide mapping and amino acid compositions in order to establish the identity of each crystallin. We have found differences in the protein structures of these two crystallins despite some degree of similarity in their amino acid compositions.
Rapid formation of secondary structure framework in protein folding studied by stopped‐flow circular dichroism
Authors: Kuwajima K. , Yamaya H. , Miwa S. , Sugai S. , Nagamura T.
Published: July 2002

Kinetic refolding reactions of ferricytochrome c and β‐lactoglobulin have been studied by stopped‐flow circular dichroism by monitoring rapid ellipticity changes of peptide backbone and side‐chain chromophores. In both proteins, a transient intermediate accumulates within the dead time of stopped‐flow mixing (18 ms), and the intermediate has an appreciable amount of secondary structure but possesses an unfolded tertiary structure. It is suggested that the rapid formation of a secondary structure framework in protein folding is a common property observed in a variety of globular proteins.
Truncated glucagon‐like peptide I, an insulin‐releasing hormone from the distal gut
Authors: Holst J.J. , Ørskov C. , Vagn Nielsen O. , Schwartz T.W.
Published: July 2002

By hydrophobic gel permeation and high pressure liquid chromatography we isolated from pig intestinal mucosa a peptide which corresponds to proglucagon 78–107 as suggested by chromatography and determination of its N‐terminal sequence. Natural and synthetic proglucagon 78–107 dose dependently and potently increased insulin secretion from the isolated perfused pig pancreas. Proglucagon 78–107 also secreted by the small intestine may participate in the hormonal control of insulin secretion.
Solute carriers involved in energy transfer of mitochondria form a homologous protein family
Authors: Aquila Heinrich , Link Thomas A. , Klingenberg Martin
Published: July 2002

The sequences of three mitochondrial carriers involved in energy transfer, the ADP/ATP carrier, phosphate carrier and uncoupling carrier, are analyzed. Similarly to what has been previously reported for the ADP/ATP carrier and the uncoupling protein, now also the phosphate carrier is found to have a tripartite structure comprising three similar repeats of approx. 100 residues each. The three sequences show a fair overall homology with each other. More significant homologies are found by comparing the repeats within and between the carriers in a scheme where the sequences are spliced into repeats, which are arranged for maximum homology by allowing possible insertions or deletions. A striking conservation of critical residues, glycine, proline, of charged and of aromatic residues is found throughout all nine repeats. This is indicative of a similar structural principle in the repeats. Hydropathy profiles of the three proteins and a search for amphipathic α‐spans reveal six membrane‐spanning segments for each carrier, providing further support for the basic structural identity of the repeats. The proposed folding pattern of the carriers in the membrane is exemplified with the phosphate carrier. A possible tertiary arrangement of the repeats and the membranespanning helices is shown. The emergence of a mitochondrial carrier family by triplication and by divergent evolution from a common gene of about 100 residues is discussed.
Primary structure of rat brain sodium channel III deduced from the cDNA sequence
Authors: Kayano Toshiaki , Noda Masaharu , Flockerzi Veit , Takahashi Hideo , Numa Shosaku
Published: July 2002

The complete amino acid sequence of a third sodium channel (designated sodium channel III) from rat brain has been deduced by cloning and sequence analysis of the cDNA. This protein is homologous in amino acid sequence and shares characteristic structural features with other sodium channels.
ADP‐ribosylation of cell membrane proteins by staphylococcal α‐toxin and leukocidin in rabbit erythrocytes and polymorphonuclear leukocytes
Authors: Kato Iwao , Noda Masatoshi
Published: July 2002

Staphylococcal α‐toxin resulted in ADP‐ribosylation of the 37 and 41 kDa proteins of a membrane preparation from rabbit erythrocytes. In the presence of 100 μM GTP, the toxin ADP‐ribosylated proteins of 54 and 59 kDa and potentiated ADP‐ribosylation of the 37 and 41 kDa forms. GTP had no effect on ADP‐ribosylation of membrane proteins in the absence of α‐toxin. Incubation of a membrane preparation of rabbit polymorphonuclear leukocytes with the S and F components of staphylococcal leukocidin resulted in ADP‐ribosylation of the 37 and 41 kDa proteins, respectively. Furthermore, the 37, 41, 54 and 59 kDa proteins were ADP‐ribosylated by leukocidin in the presence of GTP. The ADP‐ribosylation of these proteins was observed to be dependent on the incubation time and toxin dose and was abolished by prior boiling. Addition of agmatine did not attenuate ADP‐ribosylation of these proteins. These results demonstrate that staphylococcal α‐toxin and leukocidin possess ADP‐ribosyltransferase activities which are potentiated by GTP and suggest that ADP‐ribosylation reactions are responsible for development of the cytolytic activities of these staphylococcal toxins.
Bradykinin stimulates tumor necrosis factor and interleukin‐1 release from macrophages
Authors: Tiffany Carol W. , Burch Ronald M.
Published: July 2002

Bradykinin and related kinins have been implicated in the initiation and maintenance of inflammation. Cytokines appear to be the primary mediators of many inflammatory diseases. The potential ability of bradykinin to stimulate release of tumor necrosis factor and interleukin‐1 from macrophages was examined. Bradykinin stimulated release of both cytokines from P388‐D1 and RAW264.7 murine macrophages. Studies with selective agonists and antagonists suggest that cytokine release is mediated by a B1 kinin receptor.
A new superfamily of putative NTP‐binding domains encoded by genomes of small DNA and RNA viruses
Authors: Gorbalenya Alexander E. , Koonin Eugene V. , Wolf Yuri I.
Published: July 2002

Statistically significant similarity was revealed between amino acid sequences of NTP‐binding pattern‐containing domains which are among the most conserved protein segments in dissimilar groups of ss and dsDNA viruses (papova‐, parvo‐, geminiviruses and P4 bacteriophage), and RNA viruses (picorna‐, como‐ and nepoviruses) with small genomes. Within the aligned domains of 100–120 amino acid residues, three highly conserved sequence segments have been identified, i.e. ‘A’ and ‘B’ motifs of the NTP‐binding pattern, and a third, C‐terminal motif ‘C’, not described previously. The sequence of the ‘B’ motif in the proteins of the new superfamily is unusually variable, with substitutions, in some of the members, of the Asp residue conserved in other NTP‐binding proteins. The ‘C’ motif is characterized by an invariant Asn residue preceded by a stretch of hydrophobic residues. As the new superfamily included a well studied DNA and RNA helicase, T antigen of SV40, helicase function could be tentatively assigned also to the other related viral putative NTP‐binding proteins. On the other hand, the possibility of different and/or multiple functions for some of these proteins is discussed.
Identification of genetically high risk individuals to lung cancer by DNA polymorphisms of the cytochrome P45 0IA1 gene
Authors: Kawajiri Kaname , Nakachi Kei , Imai Kazue , Yoshii Akira , Shinoda Nahomi , Watanabe Junko
Published: July 2002

A good correlation was observed between enhanced lung cancer risk and restriction fragment length polymorphisms (RFLPs) of the P450IA1 gene with the restriction enzyme MspI. Genotype frequencies of 0.49 for the predominant homozygote, 0.40 for the heterozygote, and 0.11 for the homozygous rare allele were observed in a healthy population. Among lung cancer patients, the frequency of homozygous rare allele of P450IA1 gene was found to be about 3‐fold higher than that among healthy population, and this difference was statistically significant. This is the first report to identify the genetically high risk individuals to lung cancer at the gene level.
1H nuclear magnetic resonance study of the solution conformation of an antibacterial protein, sapecin
Authors: Hanzawa Hiroyuki , Shimada Ichio , Kuzuhara Takashi , Komano Hiroto , Kohda Daisuke , Inagaki Fuyuhiko , Natori Shunji , Arata Yoji
Published: July 2002

The solution conformation of an antibacterial protein sapecin has been determined by 1H nuclear magnetic resonance (NMR) and dynamical simulated annealing calculations. It has been shown that the polypeptide fold consists of one flexible loop (residues 4–12), one helix (residues 15–23), and two extended strands (residues 24–31 and 34–40). It was found that the tertiary structure of sapecin is completely different from that of rabbit neutrophil defensin NP‐5, which is homologous to sapecin in the amino acid sequences and also has the antibacterial activity. The three‐dimensional structure determination has revealed that a basic‐residue rich region and the hydrophobic surface face each other on the surface of sapecin.
The cytochrome b of the sea urchin Paracentrotus lividus is naturally resistant to myxothiazol and mucidin
Authors: Esposti M.Degli , Ghelli A. , Butler G. , Roberti M. , Mustich A. , Cantatore P.
Published: July 2002

The ubiquinol:cytochrome c reductase activity of Paracentrotus lividus mitochondria is relatively insensitive to the specific inhibitors myxothiazol and mucidin. The I
50 of myxothiazol and mucidin are three and two orders of magnitude higher, respectively, in P. lividus than in bovine heart mitochondria. The natural resistance of the P. lividus reductase to these inhibitors can be correlated with a single amino replacement, an alanine for a glycine at position 143, in the sequence of cytochrome b. This position is located in a conserved region of the molecule, believed to be important in the oxidation of ubiquinol by the reductase.
Cloning and expression of a cDNA encoding a novel human neurotrophic factor
Authors: Kaisho Yoshihiko , Yoshimura Koji , Nakahama Kazuo
Published: July 2002

A cDNA encoding a novel human neurotrophic factor (designated nerve growth factor‐2; NGF‐2) was cloned from a human glioma cDNA library using a synthetic DNA corresponding to human nerve growth factor (NGF). The cloned cDNA encodes a polypeptide composed of 257 amino acid residues including a prepro‐sequence of 138 residues and a mature region of 119 residues. The amino acid sequence of human NGF‐2 exhibits 58% similarity with that of human NGF. Conditioned medium of COS‐7 cells transfected with an expression plasmid for human NGF‐2 cDNA supported the survival of sensory neurons isolated from dorsal root ganglia of embryonic chicks. A 1.5 kb of NGF‐2 mRNA can be detected from an early development stage in rat brain, by Northern blotting analysis.
Trypsin‐SBTI interaction in reverse micelles A slow intermicellar exchange‐dependent binding
Authors: Bru Roque , Garcia-Carmona Francisco
Published: July 2002

Solubilisate exchange between reverse micelles must take place before any reaction inside reverse micelles occurs if the reactants are confined to the aqueous micellar core. When the interacting species are 2 small molecules or one small molecule and one macromolecule, it has been shown that the exchange is faster than the typical turnover of an enzymatic reaction. The study of the interaction between 2 macromolecules (trypsin and soybean trypsin inhibitor) in reverse micelles carried out in this work reveals that the exchange between these macromolecule‐containing reverse micelles slows down by a thousand times and the limiting‐step in the exchange, the fusion, by 10* times. Both reverse micellar size (ωo = [water]/ [surfactant]) and temperature affected the rate of the fusion process. A hypothesis for the proposed active role of macromolecules in the exchange process is also given.
Crystallization and preliminary X‐ray diffraction data of the Fab fragment of a monoclonal antibody against apamin, a bee venom neurotoxin
Authors: Devaux Christiane , Defendini Maria-Léria , Alzar Pedro M. , Abergel Chantal , Granier Claude , Fontecilla-Camps Juan C.
Published: July 2002

Fab fragments of anti‐apamin monoclonal antibodies have been purified to homogeneity and crystallized. The crystals belong to the monoclinic space group P21 with cell dimensions a = 99.0 ± 0.3 Å, b = 137.1 ± 0.4 Å, c = 76.0 ± 0.2 Å and β = 92.9 ± 0.9°. They most likely contain four molecules in the asymmetric unit (V
m = 2.39 Å3/Da). The possibility of the existence of non‐crystallographic symmetry is discussed.
Identification and nucleotide sequence determination of the gene responsible for Ogawa serotype specificity of V. cholerae 01
Authors: Ito Teruyo , Ohshita Yoshihiro , Hiramatsu Keiichi , Yokota Takeshi
Published: July 2002

The gene encoding a protein of 27 kDa, which is specifically expressed in Vibrio cholerae of serotype Ogawa, was identified and its nucleotide sequence determined. The plasmid carrying this gene was found to convert serotype specificity from Inaba to Ogawa when introduced into the Escherichia coli DH5(pVCI112) cell which harbors a cloned 20‐kilobase genomic DNA fragment of V. cholerae NIH35A3 and expresses the 01 antigen of Inaba serotype.
Cloning, pharmacological characteristics and expression pattern of the rat GABAA receptor α4 subunit
Authors: Wisden William , Herb Anne , Wieland Heike , Keinänen Kari , Lüddens Hartmut , Seeburg Peter H.
Published: July 2002

A cDNA of rat brain encoding the GABAA receptor α4 subunit has been cloned. Recombinant receptors composed of α4, β2 and γ2 subunits bind with high affinity the GABA agonist [3H]muscimol and the benzodiazepine ‘alcohol antagonist’ [3H]Ro 15‐4513, but fail to bind benzodiazepine agonists. The α4 subunit is expressed mainly in the thalamus, as assessed by in situ hybridization histochemistry, and may participate in a major population of thalamic GABAA receptors. The α4 mRNA is found at lower levels in cortex and caudate putamen, and is rare in cerebellum.
Cloning, expression and chromosomal localization of a new putative receptor‐like protein tyrosine phosphatase
Authors: Gebbink Martijn F.B.G. , van Etten Ingrid , Hateboer Guus , Suijkerbuijk Ron , Beijersbergen Roderick L. , van Kessell Ad Geurts , Moolenaar Wouter H.
Published: July 2002

We have isolated a mouse cDNA of 5.7 kb, encoding a new member of the family of receptor‐like protein tyrosine phosphatases, termed mRPTPμ. The cDNA predicts a protein of 1432 amino acids (not including signal peptide) with a calculated M
r of 161 636. In addition, we have cloned the human homologue, hRPTPμ, which shows 98.7% amino acid identity to mRPTPμ. The predicted mRPTPμ protein consists of a 722 amino acid extracellular region, containing 13 potential N‐glycosylation sites, a single transmembrane domain and a 688 amino acid intracellular part containing 2 tandem repeats homologous to the catalytic domains of other tyrosine phosphatases. The N‐terminal extracellular part contains a region of about 170 amino acids with no sequence similarities to known proteins, followed by one Ig‐like domain and 4 fibronectin type III‐like domains. The intracellular part is unique in that the region between the transmembrane domain and the first catalytic domain is about twice as large as in other receptor‐like protein tyrosine phosphatases. RNA blot analysis reveals a single transcript, that is most abundant in lung and present in much lower amounts in brain and heart. Transfection of the mRPTPμ cDNA into COS cells results in the synthesis of a protein with an apparent Mr
of 195 000, as detected in immunoblots using an antipeptide antibody. The human RPTPμ gene is localized on chromosome 18pter‐q11, a region with frequent abnormalities implicated in human cancer.
UV difference spectroscopy of ligand binding to maltose‐binding protein
Authors: Gehring Kalle , Bao Kogan , Nikaido Hiroshi
Published: July 2002

We have used UV absorbance spectroscopy to study the binding of linear and circular maltodextrins to maltose‐binding protein (MBP). Titrations with maltose yield three ??? points in the difference spectrum of MBP, consistent with two protein conformations: ligand‐free and ligand‐bound. In contrast, titrations with maltotetraose reveal three conformation: ligand‐free, a low‐affinity liganded state, and a high affinity liganded state. These results confirm and extend the results from tritium NMR spectroscopy, namely, that MBP can bind maltodextrin either by the sugar's anomeric end (high affinity) or by the middle of the maltodextrin chain flow affinity.
A laser flash absorption spectroscopy study of Anabaena sp. PCC 7119 flavodoxin photoreduction by photosystem I particles from spinach
Authors: Medina Milagros , Hervás Manuel , Navarro José A. , De la Rosa Miguel A. , Gómez-Moreno Carlos , Tollin Gordon
Published: July 2002

Electron transfer from P700 in photosystem I (PSI) particles from spinach to Anabaena sp. PCC 7119 flavodoxin has been studied using laser flash absorption spectroscopy. A non‐linear protein concentration dependence of the rate constants was obtained, suggesting a two‐step mechanism involving complex formation (k=3.6 × 107 M−1· s−1) followed by intracomplex electron transfer (k=270 s−1). The observed rate constants had a biphasic dependence on the concentrations of NaCl or MgCl2 with maximum values in the 40–80 mM range for NaCl and 4–12 mM for MgCl2. To our knowledge, this is the first time that the kinetics of PSI‐dependent flavodoxin photoreduction have been determined.
Lack of DNA enzymatic photoreactivation in HeLa cell‐free extracts
Authors: Chao Chuck C.-K.
Published: July 2002

Using a DNA‐protein binding assay, we have previously identified and characterized a UV‐damaged DNA recognition protein (UVDRP) from HeLa cells [(1991) Nucleic Acids Res. 19, 6413‐6418]. In this report, the photoreactivating activity of UVDRP from the yeast, Saccharomyces cerevisiae, and HeLa cells was investigated. Although yeast and human cells are evolutionarily different from each other, both UVDRPs were conserved in the sense of their biochemical characteristics except that the yeast UVDRP also exhibited an enzymatic photoreactivating activity. A mammalian expression vector plasmid DNA carrying the bacterial chloramphenicol acetyltransferase (CAT) gene was UV irradiated in vitro followed immediately by exposure to photoreactivating light, and its transient expression in repair‐deficient xeroderma pigmentosum (XP) cells was investigated. More than 80% of the CAT activity was inhibited by UV irradiation, which was partially restored (> 60%) by a partially purified yeast photolyase. In contrast, HeLa cell extracts did not express a photoreactivatable recovery from UV‐induced inhibition of the CAT activity tested in the same system. This study has demonstrated the potential use of the DNA‐mobility shift assay to investigate enzymatic photoreactivation, and has indicated the absence of the repair mechanism in human cells.
Differanisole A, an inducer of the differentiation of Friend leukemic cells, induces stalk cell differentiation in Dictyostelium discoideum
Authors: Kubohara Yuzuru , Okamoto Koji , Tanaka Yoshimasa , Asahi Ken-ichi , Sakurai Akira , Takahashi Nobutaka
Published: July 2002

Differanisole A isolated from the conditioned medium of a soil microorganism, Chaetomium strain RB‐001, is an inducer of the differentiation of the Friend leukemic cells (mouse leukemia cells). The chemical structure of this substance is very similar to that of stalk cell differentiation‐inducing factor (DIF) isolated from the cellular slime mould, Dictyostelium discoideum. We examined the effects of differanisole A on Dictyostelium HM44 cells, a mutant strain which is defective in DIF production, and found this substance to be an inducer of stalk cell differentiation in D. discoideum.
Characterisation of wild‐type and mutant forms of human monoamine oxidase A and B expressed in a mammalian cell line
Authors: Gottowik Jürgen , Cesura Andrea M. , Malherbe Parichehr , Lang Gabrielle , Da Prada Mosé
Published: July 2002

Monoamine oxidase (MAO)‐A and MAO‐B are FAD‐containing mitochondrial enzymes which catabolize biogenic and xenobiotic amines. The N‐terminal regions of both forms of MAO contain an ADP‐binding consensus sequence found in several dinucleotide‐dependent enzymes, but otherwise show remarkable sequence differences. In order to investigate whether the N‐terminal region of MAOs participates in the different catalytic properties and inhibitor specificities exhibited by MAO‐A and MAO‐B, we constructed chimeric A/B forms and expressed them in a human embryonic kidney cell line (293 cells). The MAO‐A chimeric form containing the N‐terminus (36 amino acids) of MAO‐B and the B chimera having the first 45 amino acid sequence of MAO‐A were both catalytically active. Compared to the respective wild‐type form, they did not show any significant difference in their catalytic properties (K
m, k
cat) towards the substrates tested or in their sensitivity towards inhibitors. This indicates that the N‐terminal region of the two isoenzymes is not involved in the different specificities of MAO‐A and MAO‐B. Substitution of Cys‐397 of MAO‐B, i.e. the residue covalently anchoring FAD, with an Ala or a His residue resulted in the total loss of enzymatic activity, suggesting that the covalent coupling of FAD to MAO occurs specifically at the ‐SH group of cysteine.
Switch of histamine receptor expression from H2 to H1 during differentiation of monocytes into macrophages
Authors: Wang Ke-Yong , Arima Nobuyuki , Higuchi Satoshi , Shimajiri Shohei , Tanimoto Akihide , Murata Yoshitaka , Hamada Tetsuo , Sasaguri Yasuyuki
Published: July 2002

It is known that histamine suppresses gene expression and synthesis of tumor necrosis factor alpha (TNF‐α) induced by lipopolysaccharide (LPS) in human peripheral blood mononuclear monocytes (HPM) or alveolar macrophages via histamine H2 receptors. We investigated the effect of histamine and differentiation in macrophages on the expression and secretion of TNF‐α, TNF‐α‐converting enzyme (TACE), and histamine H1 and H2 receptors by use of a leukemia cell line, U937, and HPM. Differentiation of U937 and HPM cells with 12‐O‐tetradecanoylphorbol‐13‐acetate (TPA) enhanced the H1 receptor expression and rather suppressed the H2 receptor, resulting in up‐regulation of the histamine‐induced expression and secretion of TNF‐α, modulated via TACE. Therefore, histamine failed to inhibit up‐regulated expression of TNF‐α induced by LPS in macrophages. The switch from H2 to H1 receptors during differentiation in the monocyte/macrophage lineage could participate in the pathogenic processes of atherosclerosis and inflammatory reactions in the arterial wall.
Novel fluorescent protein from Discosoma coral and its mutants possesses a unique far‐red fluorescence
Authors: Fradkov Arkady F. , Chen Ying , Ding Li , Barsova Ekaterina V. , Matz Mikhail V. , Lukyanov Sergey A.
Published: July 2002

A novel gene for advanced red‐shifted protein with an emission maximum at 593 nm was cloned from Discosoma coral. The protein, named dsFP593, is highly homologous to the recently described GFP‐like protein drFP583 with an emission maximum at 583 nm. Using the remarkable similarity of the drFP583 and dsFP593 genes, we performed a ‘shuffling’ procedure to generate a pool of mutants consisting of various combinations of parts of both genes. One ‘hybrid gene’ was chosen for subsequent random mutagenesis, which resulted in a mutant variant with a uniquely red‐shifted emission maximum at 616 nm.
Antibacterial activity of a pepsin‐derived bovine hemoglobin fragment
Authors: Froidevaux R. , Krier F. , Nedjar-Arroume N. , Vercaigne-Marko D. , Kosciarz E. , Ruckebusch C. , Dhulster P. , Guillochon D.
Published: July 2002

Peptic digestion of bovine hemoglobin yields a fragment with antibacterial activity. This peptide was purified to homogeneity by a two‐step procedure including anion exchange chromatography and preparative reversed‐phase HPLC. Mass determination and fragmentation indicated that this peptide corresponded to the 1–23 fragment of the α chain of hemoglobin. The minimum inhibitory concentration and mode of action of this peptide towardsMicrococcus luteusstrain A270 were determined. Hemolytic assay, interaction with liposomes, and study of its structure in solution were also performed.
Isoproterenol inhibits resistin gene expression through a GS‐protein‐coupled pathway in 3T3‐L1 adipocytes
Authors: Fasshauer Mathias , Klein Johannes , Neumann Susanne , Eszlinger Markus , Paschke Ralf
Published: July 2002

Resistin was recently identified as a hormone secreted by adipocytes which leads to insulin resistance in vivo and in vitro and might therefore be an important link between obesity and diabetes. To clarify the regulation of resistin gene expression, 3T3‐L1 adipocytes were treated with various agents known to modulate insulin sensitivity, and resistin mRNA was measured by quantitative real‐time reverse transcription‐polymerase chain reaction. Interestingly, isoproterenol treatment reduced the level of resistin mRNA to 20% of non‐treated control cells. This effect was dose‐dependent with significant inhibition occurring at concentrations as low as 10 nM isoproterenol. Moreover, pretreatment of adipocytes with the β‐adrenergic antagonist propranolol almost completely reversed the inhibitory effect of isoproterenol, whereas addition of the α‐adrenergic antagonist phentolamine did not have any effect. Furthermore, the effect of isoproterenol could be mimicked by activation of GS‐proteins and adenylyl cyclase. Thus, both cholera toxin and forskolin decreased resistin mRNA expression in a dose‐dependent fashion by up to 90% of control levels. Taken together, these results suggest that resistin gene expression is regulated by a protein kinase A‐dependent pathway in 3T3‐L1 adipocytes.
Subunit stoichiometry of the pancreatic β‐cell ATP‐sensitive K+ channel
Authors: Inagaki Nobuya , Gonoi Tohru , Seino Susumu
Published: July 2002

We have investigated the subunit stoichiometry of the pancreatic β‐cell ATP‐sensitive K+ (KATP) channel (SUR1/Kir6.2 channel) by constructing cDNA encoding a single polypeptide (βα polypeptide) consisting of a SUR1 (β) subunit and a Kir6.2 (α) subunit. 86Rb+ efflux and single‐channel properties of COS1 cells expressing βα polypeptides were similar to those of COS1 cells coexpressing α monomers and β monomers. Coexpression of βα polypeptides with α monomers inhibited the K+ currents, while coexpression with β monomers did not. We then constructed another single polypeptide (βα2) consisting of a β subunit and a dimeric repeat of the α subunit. 86Rb+ efflux from COS1 cells expressing βα2 polypeptides was small, but was restored by supplementation with β monomers. These results indicate that the activity of KATP channels is optimized when the α and β subunits are coexpressed with a molar ratio of 1:1. Since inward rectifier K+ channels are thought to function as homo‐ or hetero‐tetramers, this suggests that the KATP channel functions as a multimeric protein, most likely a hetero‐octamer composed of a tetramer of the Kir6.2 subunit and a tetramer of the SUR1 subunit.
ATR‐FTIR spectroscopic investigation of E. coli transconjugants β‐lactams‐resistance phenotype
Authors: Bouhedja W , Sockalingum G.D , Pina P , Allouch P , Bloy C , Labia R , Millot J.M , Manfait M
Published: July 2002

Hyphenation of attenuated total reflection Fourier transform infrared spectroscopy and cluster analysis has been used to characterise a susceptible Escherichia coli K12 strain and the transconjugants TEM‐1, TEM‐2, TEM‐3, SHV‐2, SHV‐3, SHV‐4. A good discrimination of the susceptible strain from the transconjugants was obtained. Although a limited success was achieved in the differentiation of SHV and TEM phenotypes in general, results obtained with TEM‐2 and SHV‐3 were convincing. Spectral differences observed are ascribed to the global effects of the conjugation process, particularly their repercussions in the nucleic acids and carbohydrate absorbing regions, rather than to β‐lactamase point‐mutations.