bluuebunny/crossref_metadata_2025 huggingface dataset:
Intervenção do profissional de saúde mental em situações de perda e luto no Brasil
Authors: Santos Gabriela Casellato Brown Ferreira
Published: July 2020

Luto é tido pela Psicologia como uma resposta natural e esperada diante de rupturas de vínculos. No entanto, considera-se que 10 a 25% dos enlutados  da população mundial tendem a apresentar reações de intensidade, duração  e funcionalidade consideradas comprometidas, desenvolvendo um quadro denominado “luto complicado”. Ações preventivas, suportivas e de tratamento foram empreendidas nas últimas duas décadas no Brasil e em outros países.  A partir de uma revisão bibliográfica foram identificadas as diferentes possibilidades de intervenções realizadas no Brasil e suas funções específicas no suporte psicológico ao enlutado. Uma pesquisa em bases virtuais de dados entre 2007 e 2017 demonstra que a maioria das intervenções psicológicas com enlutados ali mencionadas aconteceram no setting hospitalar e que os atendimentos em psicoterapia se deram com base em diferentes abordagens teóricas. Observa-se ainda nessas bases uma escassez de artigos sobre a eficácia das intervenções psicológicas com enlutados, aspecto que aponta para a necessidade de ampliação das pesquisas acadêmicas sobre esta questão.
O morrer: uma análise sociológica
Authors: Lima Carolina Peres de
Published: July 2020

KELLEHEAR, Allan. Uma história social do morrer. São Paulo: Editora Unesp, 2016. 538 p.
Memória, poder e religiosidade nas arenas romanas no início do Principado
Authors: Garraffoni Renata Senna
Published: July 2020

O objetivo central do artigo é enfocar os combates de gladiadores no início do Principado Romano, para discutir memória, poder e cotidiano. Em primeiro lugar analisarei o significado do termo munus funebre, e como os jogos eram organizados no período para, na sequência, refletir sobre as principais abordagens historiográficas sobre violência e morte nas arenas. Por fim, analisarei os relevos funerários e as armas dos gladiadores para propor uma abordagem cultural acerca das arenas romanas, em especial no que diz respeito às fronteiras entre a vida e a morte.
La Construcción social de la propia muerte. El suicidio y el habitus aristocrático de la elite del Imperio Romano: una Reflexión a partir de ejemplos de Plinio el Joven y Tácito
Authors: Vendramini Darío N. Sánchez
Published: July 2020

Las elites del Principado tienen una relación muy particular con el suicidio, al que consideran como una forma aceptable y hasta particularmente honorable de muerte. El objetivo de este trabajo es llevar el análisis de los suicidios aristocráticos del Principado del ámbito de las ideas filosóficas y su influencia, al de las prácticas sociales y sus condicionantes. Más precisamente, mi intención es argumentar que el proceso de construcción del modelo aristocrático de la muerte voluntaria romana, más allá de   la influencia indiscutible de la filosofía estoica, responde también a las disposiciones del habitus de la elite senatorial y ecuestre del Principado, en el cual se integra dentro de las prácticas de afianzamiento simbólico de la posición en el espacio social.
Memorial Funerário Mathias Haas: a trajetória da criação de um espaço sobre a morte e os cemitérios
Authors: Castro Elisiana
Published: July 2020

O artigo descreve a trajetória do Memorial Funerário Mathias Haas localizado em Blumenau, Santa Catarina, abordando as etapas de sua formação, desde o início de seu projeto  de  criação, em 2010, até  os dias atuais. O Memorial é o primeiro museu sobre o tema da morte e dos cemitérios do Brasil, e abrigará o acervo reunido pela família Haas, desde o início do século XX. O arquivo preservado pela família está relacionado com a trajetória de sua empresa, e é formado por artigos de funerária, ornamentos tumulares, fotografias, registros de empregados, desenhos, materiais de expediente, dentre outros. O acervo já está sendo inventariado e se destaca por sua diversidade, sendo atualmente um dos maiores acervos preservados sobre uma empresa do ramo.
La Santa Muerte: estigma e intercambio
Authors: Lomnitz Claudio W.
Published: July 2020

Este ensayo ofrece una perspectiva antropológica sobre la expansión reciente del culto de la Santa Muerte, a partir de un análisis de tres dimensiones: el manejo diverso de las marcas de la Santa Muerte como estigma; la distinción entre formas fuertes y débiles del culto, a partir de la naturaleza del pacto y lazo que se crea entre devotos y deidad; y las diversas formas de reciprocidad que se dan entre la Santa Muerte y sus devotos.
A morte lhe cai bem: reconsiderando o significado do mobiliário funerário na construção do prestígio social
Authors: Souza Camila Diogo de
Published: July 2020

Este artigo tem como objetivo discutir o papel do mobiliário funerário (ou acompanhamentos) na construção do prestígio e da memória social, a partir da apresentação de um estudo de caso específico. Os objetos depositados com os mortos, desde as abordagens teórico-metodológicas mais antigas dos estudos dos contextos funerários, constituíram o elemento central da cultura material de natureza funerária sobre o qual os pesquisadores se debruçavam para reconstruir aspectos da organização social e alcançar aspectos simbólicos dos rituais mortuários. Contudo, devido à caraterística multifacetada do registro arqueológico dos sepultamentos, as análises interdisciplinares têm possibilitado reconsiderar as relações entre os artefatos e os valores socioculturais de riqueza e prestígio, sob a ótica das diferentes abordagens interpretativas das linhas teórico-metodológicas da Arqueologia Funerária.
Voir Délos et mourir: La gestion de la mort interdite dans un sanctuaire grec à travers les sources épigraphiques et les données archéologiques
Authors: Prêtre Clarisse
Published: July 2020

La mort dans l’Antiquité grecque a déjà fait l’objet de nombreuses études, à travers l’histoire, la religion, l’archéologie ou l’anthropologie. De la mort héroïque étudiée par J.-P.Vernant aux considérations taphonomiques récemment rassemblées par J. Bradbury dans un travail collectif, en passant par les morts violentes analysées par Ph. Charlier, on remarque qu’en dépit des nombreux croisements interdisciplinaires réalisés, jamais n’a été menée aucune réelle confrontation des sources textuelles avec les autres champs d’études habituels. En nous appuyant sur l’exemple de Délos, le célèbre sanctuaire d’Apollon à l’époque hellénistique où il était interdit de mourir, nous étudierons la gestion de la mort dans cette île. Où enterrer les cadavres «illicites», comment gérer les morts accidentelles? En examinant minutieusement le vocabulaire du champ sémantique de la mort dans les sources épigraphiques déliennes et en portant une attention particulière aux témoignages funéraires encore en place, on mettra ainsi en exergue les stratégies funéraires de la cité et du sanctuaire de Délos, où même les morts obéissent à la volonté d’Apollon.
São Martins: um cemitério periférico, (des)conhecido na cidade de Bagé, Rio Grande do Sul (cronologia e morfologia, características sociais e culturais)
Authors: Cerqueira Fábio Vergara , Bastianello(†) Elaine M. Tonini
Published: July 2020

À Professora Elaine Maria Tonini Bastianello, in memoriam. Este artigo trata do Cemitério São Martins, situado na periferia, a norte da cidade de Bagé, Rio Grande do Sul. Propomos um modelo de quatro fases para compreender a cronologia de seus monumentos tumulares: 1) catacumbas  de inspiração neoclássica com tratamento caiado; 2) sepulturas caiadas com fachada em forma de templo com frontão encimado por cruz; 3) jazigos-capela de influência art déco com proteção em cirex; e 4) gavetas e a sociedade de massas. Considerando que os espaços de enterramento refletem o mundo dos vivos, procuramos entender essa necrópole como local alternativo à necrópole central e de elite, o Cemitério da Santa Casa de Caridade. O Cemitério São Martins é o espaço em que as classes econômicas não abastadas enterravam seus mortos e, ao mesmo tempo, estabeleciam laços de identidade.
Alimentando o luto: uma pesquisa sobre as comidas servidas nos velórios de Entre Rios de Minas e Belo Horizonte
Authors: Bonomo Juliana Resende
Published: July 2020

Este relato busca apresentar minha experiência no estudo da comensalidade nos velórios antigos e atuais em Entre Rios de Minas em comparação com igual prática nos velórios promovidos pelas funerárias de luxo em Belo Horizonte. Circunscrita na área de conhecimento da Memória Social, a pesquisa também dialogou com a História Cultural e a Sociologia. A partir do uso de metodologia da História Oral foi possível apreender a dimensão cultural do serviço de comidas e bebidas nos velórios antigos, além da mercantilização do morrer, promovida pela chegada das casas funerárias, que transformou essa tradição. Ao longo da pesquisa realizada foi possível identificar a ressignificação da tradição antiga de compartilhar comidas e bebidas nos velórios.
A mi hijo lo eligió dios. Un análisis sobre las moralidades en torno a muertes violentas en villa La Tela (Córdoba-Argentina)
Authors: Liberatori Marina
Published: July 2020

En este artículo busco reflexionar, etnográficamente, sobre diferentes moralidades configuradas en torno a una muerte acontecida en una villa de Córdoba. Ludo Hernández era un joven de 18 años quien era acusado de incurrir en prácticas delictivas tales como robos, consumo y venta de drogas, entre otras. Murió asesinado el 29 de diciembre de 2010. A partir de allí los vecinos de La Tela construyeron diversos relatos sobre esta muerte; muchos de ellos vinculados con cuestionamientos morales relacionados con su conducta, con la educación recibida por su familia, y por consiguiente, con la reputación de la misma. Su madre, por su parte, intentaba “limpiar” la memoria del muerto, a la vez que canalizar el sufrimiento. Intentaré reconstruir las diferentes explicaciones en torno a esta muerte, para ello compararé la muerte de Ludo Hernández, con la de otro joven quien también fue asesinado en la villa, pero en un contexto completamente diferente, puesto que era considerado por sus vecinos como un “chico bueno” y por tanto, su muerte fue llorada y considerada “injusta”.
Grupo de Apoio aos Enlutados pelo Suicídio: uma experiência de posvenção e suporte social
Authors: Scavacini Karen , Cornejo Elis Regina , Cescon Luciana França
Published: July 2020

O tabu e o estigma produzidos culturalmente em torno do suicídio impõem interdições e sanções no processo de luto dos sobreviventes do suicídio, o que implica em uma dificuldade na simbolização da perda, devido ao não reconhecimento e validação social desse tipo de morte. Estas características particulares denunciam a falha de suporte social ao sobrevivente enlutado por suicídio que pode contribuir com o intenso sofrimento vivido e para a complicação na elaboração do luto. O presente artigo tem como finalidade apresentar um relato de experiência de pósvenção do suicídio com o Grupo de Apoio aos Enlutados pelo Suicídio, abordando a importância deste espaço para a promoção de um suporte social que habitualmente é frágil ou inexistente no processo de luto destes sobreviventes por suicídio. Esta experiência evidencia o grupo de apoio como um importante espaço para elaboração do luto por suicídio, onde o pesar e o sofrimento são expressos e validados, possibilitando que os enlutados possam encontrar um lugar de fala e pertencimento entre os pares.
Biosynthesis of intestinal microvillar proteins
Authors: Danielsen E.Michael , Cowell Gillian M.
Published: July 2002

The effect of tunicamycin on synthesis and intracellular transport of pig small intestinal aminopeptidase N (EC 3.4.11.2), sucrase‐isomaltase (EC 3.2.1.48–10) and maltase‐glucoamylase (EC 3.2.1.20) was studied by labelling of mucosal explants with [35S]methionine. The expression of the microvillar enzymes was greatly reduced by tunicamycin but could be partially restored by leupeptin, suggesting the existence of a mechanism whereby newly synthesized, malprocessed enzymes are recognized and degraded. In the presence of tunicamycin, polypeptides likely to represent non‐glycosylated forms of the enzymes persisted in the Mg2+‐precipitated membrane fraction, indicating that high mannose glycosylation is essential for transport to the microvillar membrane. Treatment of aminopeptidase N and sucrase‐isomaltase with endo F reduced the size of the high mannose forms approximately to those seen in the presence of tunicamycin. The complex forms were also sensitive to endo F but did not coincide with the high mannose forms after treatment, indicating that the size difference cannot alone be ascribed to processing of N‐linked carbohydrate.
Isolation and reconstitution of the N‐formylpeptide receptor from HL‐60 derived neutrophils
Authors: Hoyle Peter C. , Freer Richard J.
Published: July 2002

A multifunctional receptor for N‐formylpeptides exists on the membranes of neutrophils. This receptor has now been isolated from neutrophils derived from HL‐60 promyelocytic leukemia cells. After solubilization by Nonidet‐P40 and purification by affinity chromatography and HPLC the isolated receptor was reconstituted into egg phosphatidylcholine vesicles by SM‐2 Bio‐Bead removal of the Nonidet‐P40. Analysis of the affinity and selectivity of the receptor was done by direct binding of two high‐affinity ligands, formyl‐Met‐Leu‐[3H]Phe‐OH and formyl‐Nle‐Leu‐Phe[3H]Tyr‐OH. The data suggest that the receptor can be isolated and reconstituted without apparent alteration of its binding affinity and selectivity, and that there appear to be no co‐factors or subunits upon which these binding characteristics are dependent.
Kringles: modules specialized for protein binding
Authors: Patthy L. , Trexler M. , Váli Z. , Bányai L. , Váradi A.
Published: July 2002

Prothrombin, plasminogen, urokinase‐ and tissue‐type plasminogen activators contain homologous structures known as kringles. The kringles correspond to autonomous structural and folding domains which mediate the binding of these multidomain proteins to other proteins. During evolution the different kringles retained the same gross architecture, the kringle‐fold, yet diverged to bind different proteins. We show that the amino acid sequences of the type II structures of the gelatin‐binding region of fibronectin are homologous with those of the protease‐kringles. Prediction of secondary structures revealed a remarkable agreement in the positions of predicted β‐sheets, suggesting that the folding of kringles and type II structures may also be similar. As a corollary of this finding, the disulphide‐bridge pattern of type II structures is shown to be homologous to that in kringles. It is noteworthy that protease‐kringles and fibronectin type II structures have similar functions inasmuch as they mediate the binding of multidomain proteins to other proteins. It is proposed that the kringles of proteases and type II structures of fibronectin evolved from a common ancestral protein binding module.
Resonance Raman evidence of chloride binding to the heme iron in myeloperoxidase
Authors: Ikeda-Saito Masao , Argade Pramod V. , Rousseau Denis L.
Published: July 2002

The resonance Raman spectra of ferric derivatives of myeloperoxidase at pH 8 show ligand‐dependent differences. The data are consistent with the resting enzyme and the chloride and fluoride derivatives all having 6‐coordinated high‐spin configurations. At pH 4 we find that the resting enzyme is susceptible to photodegradation from our low power incident laser beam. Chloride binding inhibits this denaturation. Our data support direct binding of chloride to the enzyme under physiological conditions.
Quantitative measurement of the total, peroxyl radical‐trapping antioxidant capability of human blood plasma by controlled peroxidation
Authors: Wayner D.D.M. , Burton G.W. , Ingold K.U. , Locke S.
Published: July 2002

Plasma obtained from fasted humans has been analysed for total radical‐trapping antioxidant content by subjecting it to controlled peroxidation using the thermal decomposition of water‐soluble azobis (2‐amidinopropane hydrochloride) at 37°C to produce peroxyl radicals at a known, steady rate. It is found that the total radical‐trapping antioxidant content is rather similar for the 7 subjects that have been tested and, furthermore, it is 10–20‐times larger than the effect attributable to vitamin E alone. Although it is shown that urate and ascorbate augment the contribution from vitamin E, their contributions (21–34 and 0–2%, respectively) still leave 57–73% of the total antioxidant content unaccounted for. Evidence is presented to show that this previously unrecognized large reserve of antioxidant capacity is attributable to the plasma proteins.
Application of monoclonal antibodies to the isolation and characterization of a killer toxin secreted by Hansenula mrakii
Authors: Yamamoto Tetsuro , Imai Mitsunobu , Tachibana Katsumi , Mayumi Makoto
Published: July 2002

A strain of yeast, Hansenula mrakii, secretes a toxin that kills sensitive yeasts, such as Saccharomyces cerevisiae. Monoclonal antibodies raised against the toxin had both binding and neutralizing activities. The toxin in culture media was isolated by an affinity column of monoclonal antibody. The toxin is a basic polypeptide with an isoelectric point at pH 9.1, and devoid of mannosides. It is composed of 88 amino acid residues with a molecular size of 10721 Da. The monoclonal antibodies could be applicable to the analysis of biologically active sites on the toxin, in an attempt to synthesize chemically a small peptide with killer activity and little immunogenicity.
Amiloride selectively inhibits the urokinase‐type plasminogen activator
Authors: Vassalli Jean-Dominique , Belin Dominique
Published: July 2002

The diuretic drug amiloride, an inhibitor of Na+ uptake, competitively inhibits the catalytic activity of the urokinase‐type plasminogen activator (u‐PA), with a K
i of 7 × 10−6 M. Generation of plasmin, cleavage of peptide substrates, and interaction of u‐PA with a specific macromolecular proteinase inhibitor are all prevented in the presence of the drug. In contrast, amiloride does not affect the activity of either tissue‐type plasminogen activator, plasmin, plasma kallikrein or thrombin. The inhibition of u‐PA by amiloride may be related to the previously reported inhibition of u‐PA‐type enzymes by Na+. Amiloride or related compounds could prove useful in selectively controlling u‐PA‐catalyzed extracellular proteolysis.
Intracellular organelle motility and membrane fusion processes in human neutrophils upon cell activation
Authors: Mollinedo Faustino , Schneider Donald L.
Published: July 2002

Release subcellular fractionation experiments indicate that fusion of a novel tertiary granule with the plasma membrane is concomitant with human neutrophil activation. Phorbol 12‐myristate 13‐acetate (PMA) induced a respiratory burst in human neutrophils as well as a high release of gelatinase, a marker of the tertiary granule. Preincubation of neutrophils with cytochalasin E induced a partially activated or ‘primed’ state, in which cells were unable to generate superoxide anion, but showed a reduced latency period for this activity. Fusion of tertiary granules with the cell surface also occurred during priming, although to a lesser extent than in PMA stimulation. The rapid tertiary granule degranulation, preceding that of specifics azurophilics, seems to play an important role in the functionality and secretory properties of human neutrophils.
Low‐temperature spectrophotometry of phoborhodopsin
Authors: Shichida Yoshinori , Imamoto Yasushi , Yoshizawa Tôru , Takahashi Tetsuo , Tomioka Hiroaki , Kamo Naoki , Kobatake Yonosuke
Published: July 2002

Photoreactions of the fourth rhodopsin‐like pigment, phoborhodopsin (pR480), ofHalobacterium halobiumwere studied by low‐temperature spectrophotometry. Upon irradiation of pR480 at ‐ 80 as well as ‐ 170°C with 436 nm light, a new intermediate having a difference absorption maximum at about 520 nm was produced. We designate this intermediate 'P520'. Prolonged irradiation caused the formation of a photosteady‐state mixture composed of P520 and pR480, indicating that P520 was photoreversible to pR480 at ‐ 80°C. P520 was thermally converted back to pR480 through P350 and P530, both of which were detected previously [(1986) Biochem. Biophys. Res. Commun. 139, 389–395]. Based on these results, a new photochemical reaction cycle of pR is presented.
Endothelin stimulates c‐fos and c‐myc expression and proliferation of vascular smooth muscle cells
Authors: Komuro Issei , Kurihara Hiroki , Sugiyama Takao , Takaku Fumimaro , Yazaki Yoshio
Published: July 2002

Recently, a potent vasoconstrictor peptide, endothelin (EDT), was isolated from vascular endothelial cells. We examined its effect on rat vascular smooth muscle cells (VSMCs). EDT induced the elevation of intracellular calcium, which was dependent on extracellular calcium and inhibited by a calcium‐channel antagonist in a competitive manner. EDT caused a rapid and transient increase in the c‐fos and c‐myc mRNA levels and stimulated the DNA synthesis of VSMCs in a dose‐dependent manner. This effect of EDT on the proliferation of VSMCs might be related to the development of atherosclerosis.
Demonstration of a high density lipoprotein (HDL)‐binding protein in Hep G2 cells using colloidal gold‐HDL conjugates
Authors: Kambouris Ambrosios M. , Roach Paul D. , Nestel Paul J.
Published: July 2002

Solubilized membrane proteins of Hep G2 cells were electrophoretically separated on polyacrylamide gels and electrotransferred onto nitrocellulose paper. Overlaying the nitrocellulose with human high density lipoproteins conjugated to colloidal gold revealed the presence of a single protein band with an apparent molecular mass of 80 kDa. Binding of the conjugates to this protein was specific for high density lipoproteins in as much as it was effectively displaced by an excess of unlabelled high density lipoproteins but not by a similar excess of unlabelled low density lipoproteins. Binding was not dependent on Ca2+ as 10 mM EDTA had no effect. The binding activity of the solubilized membranes was increased by incubating the cells with non‐lipoprotein cholesterol. This was detected on electroblots and quantified with a new dot blot assay using the colloidal gold‐high density lipoprotein conjugates.
Evidence for the photo‐induced oxidation of the primary electron donor P680 in the isolated photosystem II reaction centre
Authors: Telfer Alison , Barber James
Published: July 2002

A steady‐state reversible signal peaking at 680 nm in the red region of the light‐dark difference spectrum has been generated in the isolated D1/D2 photosystem II reaction centre using silicomolybdate as an electron acceptor. The signal is dependent on the structural integrity of the reaction centre and is probably due to the formation of P680+resulting from oxidation of the primary donor, thought to be a chlorophylladimer. The spectral shape of the red band of the difference spectra, however, contains a shoulder at 672 nm which suggests that monomeric chlorophyll within the reaction centre may also be photooxidised. Prolonged illumination of the sample with bright white light causes a selective bleaching of the long‐wavelength absorbing form of chlorophyll within the reaction centre. Protection against this photodestruction of P680 occurs when silicomolybdate is present during the preillumination.
DNA polymerase I (Klenow fragment): Role of the structure and length of a template in enzyme recognition
Authors: Kolocheva T.I. , Nevinsky G.A. , Volchkova V.A. , Levina A.S. , Khomov V.V. , Lavrik O.I.
Published: July 2002

The values of K
d and Gibbs energy (ΔG°) have been measured for complexes of the template site of DNA polymerase I Klenow fragment with the homo‐oligonucleotides d(pC)
n
, d(pT)
n
, d(pG)
n
and d(pA)
n
and hetero‐oligonucleotides of various structures and lengths. These parameters were evaluated from the protective effect of the oligonucleotide on enzyme inactivation by the affinity reagents d(Tp)2C[Pt2+(NH3)2OH](pT)7 and d[(Tp)2C(Pt2+(NH3)2OH)p]3T of the template site. The present results and previously reported data [(1985) Biorg. Khim. 13, 357–369] indicate that the nucleoside components of the template form complexes as a result of their hydrophobic interactions with the enzyme. Only one template internucleotide phosphate forms an Me2+‐dependent electrostatic contact and a hydrogen bond with the enzyme. The 19–20‐nucleotide fragments of the template appear to interact with the protein molecule.
Low‐molecular‐mass proteins in cyanobacterial photosystem II: Identification ofpsbH andpsbK gene products by N‐terminal sequencing
Authors: Koike Hiroyuki , Mamada Kaoru , Ikeuchi Masahiko , Inoue Yorinao
Published: July 2002

The O2‐evolving photosystem II core complex was isolated from a thermophilic cyanobacterium,Synechococcus vulcanusCopeland. Analysis by SDS‐polyacrylamide gel electrophoresis revealed that the complex contained at least seven low‐molecular‐mass proteins in addition to the well characterized CP47 apoprotein, CP43 apoprotein, 33 kDa extrinsic protein, D1 protein, D2 protein and large subunit of cytochromeb‐559. The separation profiles of these low‐molecular‐mass proteins were very similar between cyanobacterial and higher plant PS II. N‐terminal sequences of the 6.5 kDa and 3.9 kDa proteins of cyanobacterial core complex were determined after blotting to a polyvinylidene difluoride membrane. The sequence of the 6.5 kDa protein showed high homology with an internal sequence of plantpsbH gene product, so‐called 10 kDa phosphoprotein, but did not conserve the Thr residue which is specifically phosphorylated in plants. The sequence of the 3.9 kDa protein corresponded to the K protein of higher plants (mature form ofpsbK gene product). These results indicate that the products of bothpsbH andpsbK genes are present in cyanobacterial PS II as well as being associated with the O2‐evolving core complex.
α‐ and β‐tubulin mRNAs of Trypanosoma cruzi originate from a single multicistronic transcript
Authors: Soares Célia M.A. , de Carvalho Elizeu F. , Urményi Turán P. , O. Carvalho José Francisco , de Castro Firmino T. , Rondinelli Edson
Published: July 2002

The cluster of alternated α‐ and β‐tubulin genes in the genome of Trypanosoma cruzi was shown to be transcribed into a single RNA molecule which upon processing gives rise to the mature α‐ and β‐tubulin mRNAs. This conclusion was based on: (i) nuclear RNA species with the same molecular mass hybridize to both α‐ and β‐tubulin cDNA probes; (ii) S1 nuclease assay of the clustered tubulin genes has shown protected DNA fragments of the same size and of greater molecular mass than that corresponding to the mRNAs, hybridizable to both α‐ and β‐tubulin cDNA probes; (iii) β‐tubulin hybrid selected RNA is still able to hybridize to α‐tubulin probe.
Recombinant H‐chain ferritins: Effects of changes in the 3‐fold channels
Authors: Treffry Amyra , Harrison Pauline M. , Luzzago Alessandra , Cesareni Gianni
Published: July 2002

Human H‐chain ferritins bearing sequence changes in the 3‐fold channels have been expressed in E. coli to investigate the role of these channels in iron‐storage processes. The proteins assemble into shells resembling those of native ferritins. Iron uptake measurements indicate that residues in the 3‐fold channels are involved neither in initial Fe(II)‐oxidation nor in iron‐core nucleation.
Characterization of two different peptides from the venom of the scorpion Buthus sindicus
Authors: Fazal Annul , Beg Obaid U. , Shafqat Jawed , Zaidi Zafar H. , Jörnvall Hans
Published: July 2002

Two disulfide‐rich, low‐molecular mass peptides (~3 kDa and ~4 kDa) have been isolated from Buthus sindicus venom using ion‐exchange and reverse‐phase HPLC. Peptide I has 35 residues with 8 half‐cystine residues and is clearly related to four‐disulfide core proteins of the neurophysin type and to toxins of other scorpion species (55–63% residue identity). Peptide II, present in low yield, has 28 residues with 6 half‐cystine residues and a structure largely dissimilar from that of peptide I and other characterized toxins, although probably still a member of the disulfide core peptide type. Consequently, scorpion venom contains, in addition to toxins characterized before, toxin‐like compounds with distant relationships.
Tropomyosin‐troponin complex stabilizes the pointed ends of actin filaments against polymerization and depolymerization
Authors: Weigt Christiane , Schoepper Beate , Wegner Albrecht
Published: July 2002

In striated muscle the pointed ends of polar actin filaments are directed toward the center of the sarcomer. Formed filaments keep a constant length of about 1 μm. As polymerization and depolymerization at free pointed ends are not sufficiently slow to account for the constant length of the filaments, we searched for proteins which occur in sarcomers and can stabilize the pointed ends of actin filaments. We observed that tropornyosintroponin complex reduces the rate of association and dissociation of actin molecules at the pointed ends more than 30‐fold. On the average, every 600 s one association or dissociation reaction has been found to occur at the pointed ends near the critical actin monomer concentration.
Photosystem I reaction‐centre proteins contain leucine zipper motifs
Authors: Webber Andrew N. , Malkin Richard
Published: July 2002

The photosystem I (PS I) reaction‐centre polypeptides, encoded by thepsaA andpsaB genes, are shown to contain several highly conserved leucine repeats, consisting of a leucine residue every seventh amino acid, similar to the leucine zipper motifs known to mediate DNA‐binding polypeptide dimerisation. In each of the PSI reaction‐centre subunits the leucine zipper motif precedes highly conserved cysteine residues which have been proposed to ligate the interpolypeptide [4Fe‐4S] centre, Fx. We propose that PS I reaction‐centre dimerisation and [4Fe‐4S] centre formation are mediated through the leucine zipper.
Chemoattractant‐induced membrane hyperpolarization in Dictyostelium discoideum A possible role for cyclic GMP
Authors: Van Duijn Bert , Wang Mei
Published: July 2002

Membrane potentials of the cellular slime mold Dictyostelium discoideum were monitored after chemotactic stimulation by measuring the distribution of the lipophilic cation tetraphenylphosphonium. Stimulation with the chemoattractant cAMP induces a transient membrane hyperpolarization which reaches its most negative value between 1 3 min after stimulation. This hyperpolarization is consistent with the opening of potassium channels. Measurements in streamer F mutant cells reveal that cGMP likely plays a role in the regulation of the cAMP‐induced hyperpolarization.
The primary structure of the larger subunit of soluble guanylyl cyclase from bovine lung Homology between the two subunits of the enzyme
Authors: Koesling Doris , Harteneck Christian , Humbert Peter , Bosserhoff Armin , Frank Rainer , Schultz Günter , Böhme Eycke
Published: July 2002

The primary structure of the larger subunit of the soluble guanylyl cyclase from bovine lung, which catalyzes the formation of cyclic GMP from GTP, has been determined. Two clones, isolated from two bovine libraries yielded a total of 3261 bp with a coding region of 2073 bp. The open reading frame encodes a protein of 691 amino acids and a molecular mass of 77 500. The deduced amino acid sequence reveals regions which are, to a large extent, homologous to the sequence of the smaller subunit of the enzyme as well as to the sequences of other gyanylyl and adenylyl cyclases.
B—Z transition of poly(dG‐m5dC) induced by binding of Lys‐containing peptides
Authors: Takeuchi Hideo , Hanamura Nobuyuki , Hayasaka Hitoshi , Harada Issei
Published: July 2002

Effects of oligopeptides containing Lys residues on the conformation of poly(dG‐m5dC) have been investigated by circular dichroism spectroscopy. Lys‐Ala‐Lys (KAK) and its longer analogs with Lys‐Ala repeats are found to convert the B‐form polynucleotide to the Z form very efficiently. The ability to induce the B‐Z transition is characteristic of alternating Lys‐Ala sequences and increases exponentially with increasing number of the repeats. The heptupeptide KAKAKAK has an ability comparable with that of spermine, one of the most effective inducers hitherto known. The present results provide the list example of the B‐Z transition of poly(dG‐m5dC) induced by peptide binding.
Downregulation of protein kinase C‐γ is independent of a functional kinase domain
Authors: Freisewinkel Ina , Riethmacher Dieter , Stabel Silvia
Published: July 2002

Prolonged activation of protein kinase C (PKC) types α and β by tumor‐promoting phorbol esters leads to desensitization of the phorbol ester response, downregulation of protein kinase C activity and depletion of the protein kinase C polypeptide. When the γ isoenzyme of PKC is transiently expressed in COS‐1 cells and exposed to phorbol esters, PKC‐γ is downregulated in COS cells although these cells do not normally express this subtype. A point mutation in the purative ATP‐binding site (Lys‐380→Met‐380) of the protein kinase C γ isoenzyme which results in a kinase‐deficient enzyme does not interfere with this downregulation. Our results suggest that autophosphorylation or constitutive signalling through the protein kinase C‐γ kinase domain is not a prerequisite for downregulation of PKC activity.
Properties of the tungsten‐substituted molybdenum formylmethanofuran dehydrogenase from Methanobacterium wolfei
Authors: Schmitz R.A. , Albracht S.P.J. , Thauer R.K.
Published: July 2002

In Methanobacterium wolfei two formylmethanofuran dehydrogenases are present, one of which is a molybdenum‐ and the other a tungsten enzyme. We report here that also the ‘molybdenum’ enzyme contained tungsten when the archaeon was grown on molybdenum‐deprived medium supplemented with tungstate (1 μM). Unexpectedly the tungsten‐substituted molybdenum enzyme was catalytically active and displayed a rhombic EPR signal which was attributed to tungsten by the characteristic 113W splitting.
Molecular cloning of the full‐length cDNA encoding the human calbindin‐D9k
Authors: Jeung Eui-Bae , Krisinger John , Dann Jamie L. , Leung Peter C.K.
Published: July 2002

The full‐length cDNA encoding the human calbindin‐D9k (CaBP‐9k) has been cloned using reverse transcription/PCR methodology with rat‐ and bovine‐derived primers and intestinal RNA. A core product, and both a 5′and 3′ product encompassing the full‐length cDNA were obtained. The clones include coding region for 79 amino acids, 57 nucleotides 5′‐ and 159 nucleotides 3′‐non‐coding region, and a poly(A) tail. The deduced protein sequence is homologous Lo other mammalian CaBPs. Northern analysis revealed the mRNA in human duodenum to be about 600 nucleotides in length, Expression levels in adult human tissue were substantially lower than in child, rat or porcine intestine.
Purification of the 260 kDa cytosolic complex involved in the Superoxide production of guinea pig neutrophils
Authors: Someya Akimasa , Nagaoka Isao , Yamashita Tatsuhisa
Published: July 2002

A 260 kDa cytosolic complex (SP‐1) was purified from guinea pig neutrophils. SP‐1 was composed of 63 kDa, 47 kDa and 39 kDa proteins. The 63 kDa and 47 kDa proteins proved to correspond to human p67phox and p47phox by Western blot analysis, whereas Western blot and amino acid sequence analyses revealed that the 39 kDa protein was a novel protein. The 47 kDa protein was separated from the 63 kDa and 39 kDa proteins by dithiothreitol (DTT)‐treatment. On the other hand, the 63 kDa and 39 kDa proteins were not separated with DTT, detergent and ethanol treatment. These results suggest that the 39 kDa protein tightly associates with the 63 kDa protein and may regulate the function of the 63 kDa protein.
Metal as a novel type of the enzyme substrate
Authors: Shumilin I.A. , Nikandrov V.V. , Krasnovsky A.A. , Popov V.O.
Published: July 2002

The process of NAD+photoreduction under the coupled action of CdS semiconductor and NAD‐dependent hydrogenase from hydrogen‐oxidizing bacteriumAlcaligenes eutrophusmay be divided into light and dark stages. At the first stage, illumination of the system leads to the photooxidation of the sacrificial electron donor and results in the reduction of the semiconductor surface. At the second dark stage NAD+is reduced to NADH in the presence of hydrogenase. Atoms of metallic Cd(0) are shown to be the true substrate of the enzymatic reaction. The prerequisite for the electron transfer from Cd(0) to hydrogenase is enzyme adsorption on the semiconductor surface, the redox center of the hydrogenase reacting with Cd(0) atoms resides on the flavin‐containing heterodimer of the protein. The activity of the hydrogenase immobilized on CdS in the reaction of NAD+reduction by metallic Cd is close to the enzyme activity with the physiological substrates in solution. Thus, the first example of a metal being the substrate of the enzymatic process is presented.
CoA esters of valproic acid and related metabolites are oxidized in peroxisomes through a pathway distinct from peroxisomal fatty and bile acyl‐CoA β‐oxidation
Authors: Vamecq Joseph , Vallee Louis , Fontaine Monique , Lambert Didier , Poupaert Jacques , Nuyts Jean-Pierre
Published: July 2002

In rat liver homogenates fortified with the appropriate cofactors (ATP and CoA), valproic acid induced H2O2 production rates by far lower than those recorded on the straight medium‐chain fatty acid n‐octanoic acid. Using directly the CoA esters of these carboxylic acids as substrates for the rat liver H2O2‐generating enzyme activities, valproyl‐CoA, and n‐octanoyl‐CoA were found to induce similar oxidation rates. In the rat liver homogenates, cyanide‐insensitive valproyl‐CoA and octanoyl‐CoA oxidations occurred at rates similar to those of valproyl‐CoA and octanoyl‐CoA oxidase(s), respectively. Studies on fractions obtained from rat liver postnuclear supernatants by isopycnic centrifugation on a linear sucrose density gradient disclose that the density distribution of valproyl‐CoA oxidase superimposes to those of catalase, fatty acyl‐CoA oxidase and cyanide‐insensitive fatty acyl‐CoA oxidation, three peroxisomal marker activities. By contrast, the cyanide‐insensitive valproyl‐CoA oxidation does not adopt the typical peroxisomal distribution of these activities but rather exhibits a mitochondrial localization with, however, a minor peroxisomal component. Interestingly enough, the comparative study of rat tissue distribution, inducibility by clofibrate and sensitivity to deoxycholate indicated that valproyl‐CoA oxidase is an enzyme distinct from fatty acyl‐CoA oxidase and bile acyl‐CoA oxidase. Taken as a whole, the results presented here support the occurrence of a peroxisomal oxidation of the CoA ester of valproic acid and its Δ4‐enoic derivate which might be characterized by two major features: initiation by an acyl‐CoA oxidase distinct from fatty and bile acyl‐CoA oxidases, and inability to complete the β‐oxidation cycle which would not proceed, at significant rates, further than the β‐hydroxyacyl‐CoA dehydrogenation step in peroxisomes.
Mitochondrial carrier proteins
Authors: Palmieri F.
Published: July 2002

Ten mitochondrial carriers have been purified from animal mitochondria. They are small proteins with a molecular mass ranging from 28 to 34 kDa on SDS‐PAGE. So far, five of these proteins have been sequenced. Their polypeptide chain consists of three tandemly related sequences of about 100 amino acids. The repeats of the different proteins are related and probably fold into two transmembrane a‐helices linked by an extra‐membrane loop. The features of this family are also present in several proteins of unknown function characterized by DNA sequencing. Isoforms of some carriers have been found. All mitochondrial carriers investigated in proteoliposomes function according to a simultaneous (sequential) mechanism of transport. The only exception is the carnitine carrier that proceeds via a ping‐pong mechanism. Three mitochondrial carriers have been expressed in yeast and two overexpressed in E. coli and refolded in active form.
Vacuolar ATPase inactivation blocks recycling to the trans‐Golgi network from the plasma membrane
Authors: Reaves Barbara , Banting George
Published: July 2002

TGN38/41 is an integral membrane protein which recycles between the trans‐Golgi network (TGN) and the cell surface but is predominantly located in the TGN of rat (NRK) cells at steady state. As part of our studies on the mechanism and route of recycling between the TGN and the cell surface we have used chloroquine or Bafilomycin A1 to modulate the lumenal pH of endocytic organelles. The data we present demonstrate that inactivation of the proton pump which maintains the acidic environment within the lumen of endocytic organelles leads to an accumulation of TGN38/41 in early endosomes. These data confirm the observation that TGN38/41 recycles between the plasma membrane and the TGN and identifies a specific block in that recycling pathway.
Evidence for nitric oxide‐mediated oxidative damage in chronic inflammation Nitrotyrosine in serum and synovial fluid from rheumatoid patients
Authors: Kaur Harparkash , Halliwell Barry
Published: July 2002

Reaction of nitric oxide (NO•) with superoxide radical generates peroxynitrite, which can decompose to products that nitrate aromatic amino acids. Such nitro‐aromatics may be ‘markers’ of NO•‐dependent oxidative damage. Blood serum and synovial fluid from patients with the inflammatory joint disease rheumatoid arthritis contain 3‐nitrotyrosine. By contrast, body fluids from normal subjects and patients with osteoarthritis contain no detectable 3‐nitrotyrosine; much lower levels were found in serum from patients in the early stages of rheumatoid arthritis. This is evidence that NO• plays a role in joint damage in rheumatoid arthritis.
Molecular cloning of rat kynurenine aminotransferase: Identity with glutamine transaminase K
Authors: Mosca Monica , Cozzi Liviana , Breton Jerome , Speciale Carmela , Okuno Etsuo , Schwarcz Robert , Benatti Luca
Published: July 2002

The enzyme kynurenine aminotransferase (KAT) catalyses the conversion of l‐kynurenine to kynurenic acid. A combination of polymerase chain reaction techniques and hybridization screening was used to isolate a cDNA clone encompassing the entire coding region of KAT from rat kidney. Identification of the cDNA as coding for KAT was based both on the comparison of amino acid sequences obtained from purified rat KAT and on the expression of KAT activity in COS‐1 cells transfected with the cDNA. RNA blot analysis indicated that KAT mRNA is widely expressed in rat tissues. Cultured cells transfected with the cDNA for KAT also showed glutamine transaminase K activity. Based mainly on sequence data, these results demonstrate that rat kidney KAT is identical with glutamine transaminase K.
Level of ATP synthase activity required for yeast Saccharomyces cerevisiae to grow on glycerol media
Authors: Mukhopadhyay Arindam , Uh Misook , Mueller David M.
Published: July 2002

Two independent cold‐sensitive pet mutants in the gene (ATP5) coding for the oligomycin sensitivity conferring protein (OSCP) have been isolated in the yeast Saccharomyces cerevisiae. The mutations in both strains alter the initiating methionine codon in the ATP5 gene: ATG to ATA (Ile) and AAG (Lys). Western blot analysis of total yeast protein after the cells were grown at 18°C, 30°C, and 37°C, indicates that the level of OSCP decreased 80% relative to the wild type strain. In addition, the level of the oligomycin‐sensitive ATPase decreased 85% relative to the wild type strain, after growth at 30°C. These findings indicate that for S. cerevisiae, the level of oxidative phosphorylation can decrease 85% without showing a large growth defect on media containing glycerol at 30°C, but not at 18°C.
Comparative analysis of catalases: spectral evidence against heme‐bound water for the solution enzymes
Authors: Andersson Laura A. , Johnson Anna K. , Simms Melissa D. , Willingham Timothy R.
Published: July 2002

A recent X‐ray structural analysis of M. luteus catalase indicates heme‐bound H2O trans to the proximal tyrosinate ligand, a finding in contrast to previous X‐ray data reporting a 5‐coordinate heme for bovine liver catalase. The presence of heme‐bound H2O, requiring displacement prior to substrate‐binding, is likely to be catalytically significant for catalases. We have used magnetic circular dichroism (MCD) spectroscopy, a highly accurate method for assignment of heme spin‐ and coordination‐states, to study native, solution forms of bovine liver, M. luteus, and A. niger catalases. All three enzymes display similar spectral features with the weak (∼5 Δϵ
m [moles·cm·Tesla]−1 intensity typical of a 5‐coordinate high‐spin ferric heme. No evidence for H2O‐ligation, inducing a 6‐coordinate heme, occurred upon variation of pH or buffer composition. Therefore, we suggest that the catalytically significant structure of catalases has an unoccupied heme binding site trans to the proximal tyrosinate heme ligand.
Cloning of a potential cytochrome P450 from the Archaeon Sulfolobus solfataricus
Authors: Wright Rhonda L. , Harris Kathryn , Solow Barbara , White Robert H. , Kennelly Peter J.
Published: July 2002

A gene, CYP119, for a potential cytochrome P450 has been isolated and sequenced from the extreme acidothermophilic archaeon Sulfolobus solfataricus. The gene predicts a polypeptide of 368 amino acids containing the consensus hemebinding sequence Phe‐Gly‐Xaa‐Gly‐Xaa‐His‐Xaa‐Cys‐Xaa‐Gly‐Xaa3‐Ala‐Arg‐Xaa‐Glu. It most closely resembles the cytochrome P450s found in the bacterium Bacillus subtilis, with which it shares 129 identical amino acid residues (35%). This first sequence of a potential archaeal cytochrome P450 represents an important step in tracing the complex evolutionary history of this biologically important enzyme family.
Crystal structure of a caricain D158E mutant in complex with E‐64
Authors: Katerelos Nikolaos A. , Taylor Mark A.J. , Scott Mandy , Goodenough Peter W. , Pickersgill Richard W.
Published: July 2002

The structure of the D158E mutant of caricain (previously known as papaya protease omega) in complex with E‐64 has been determined at 2.0 Å resolution (overall R factor 19.3%). The structure reveals that the substituted glutamate makes the same pattern of hydrogen bonds as the aspartate in native caricain. This was not anticipated since in the native structure there is insufficient room to accommodate the glutamate side chain. The glutamate is accommodated in the mutant by a local expansion of the structure demonstrating that small structural changes are responsible for the change in activity.
Molecular cloning of cDNA for nonhepatic mitochondrial arginase (arginase II) and comparison of its induction with nitric oxide synthase in a murine macrophage‐like cell line
Authors: Gotoh Tomomi , Sonoki Takashi , Nagasaki Akitoshi , Terada Kazutoyo , Takiguchi Masaki , Mori Masataka
Published: July 2002

Arginase exists in two isoforms. Liver‐type arginase (arginase I) is expressed almost exclusively in the liver and catalyzes the last step of urea synthesis, whereas the nonhepatic type (arginase II) is expressed in extrahepatic tissues. Arginase II has been proposed to play a role in down‐regulation of nitric oxide synthesis. A cDNA for human arginase II was isolated. A polypeptide of 354 amino acid residues including the putative NH2‐terminal presequence for mitochondrial import was predicted. It was 59% identical with arginase I. The arginase II precursor synthesized in vitro was imported into isolated mitochondria and proteolytically processed. mRNA for human arginase II was present in the kidney and other tissues, but was not detected in the liver. Arginase II mRNA was coinduced with nitric oxide synthase mRNA in murine macrophage‐like RAW 264.7 cells by lipopolysaccharide. This induction was enhanced by dexamethasone and dibutyryl cAMP, and was prevented by interferon‐γ. Possible roles of arginase II in NO synthesis are discussed.