bluuebunny/crossref_metadata_2025 huggingface dataset:
Perancangan Proses Remanufaktur pada Komponen Otomotif
Authors: Tanoto Yopi Yusuf
Published: February 2021

Kendaraan bermotor sebagai alat transportasi saat ini menjadi salah satu kebutuhan penting untuk manusia. Jumlah sepeda motor terus meningkat setiap tahunnya. Hal ini menyebabkan sampah atau limbah berupa komponen yang telah rusak dari sebuah kendaraan juga terus meningkat. Saat ini, beberapa bentuk dari limbah tersebut dapat dilakukan recycle agar materialnya dapat dimanfaatkan kembali. Beberapa komponen yang telah rusak sebenarnya masih dapat digunakan lagi dengan beberapa proses seperti repair, refurbish atau remanufaktur. Proses remanufaktur adalah salah satu recovery option paling tinggi dalam mengolah limbah atau sampah. Dalam melakukan perancangan proses remanufaktur untuk komponen otomotif, perlu adanya pemilihan komponen yang akan dijadikan objek. Setelah pemilihan objek, maka dibuat serangkaian proses yang harus ada di dalam proses remanufaktur komponen otomotif seperti disassembly, cleaning, inspection, recondition, dan reassembly. Tahap akhirnya adalah membuat gambar rancangan proses remanufaktur untuk objek komponen otomotif. Ciri khas yang terdapat pada proses remanufaktur komponen otomotif adalah adanya special treatment seperti normalizing, hardening untuk beberapa komponen otomotif yang berguna untuk mengembalikan atau bahkan meningkatkan performa atau mechanical properties dari komponen yang dilakukan remanufaktur.
Surfactant in SARS-CoV-2 - a therapeutic option based on underlying lung cell damage?
Authors: Carvalho Sousa Maristella de Araújo , Matos de Oliveira Gabriela Correia , De Oliveira Andrade Luis Jesuino
Published: February 2021

Introduction: the severe acute respiratory syndrome – coronavirus 2 (SARS Cov-2), leads to a diffuse alveolar deterioration due infection of type II pneumocytes. The type II pneumocytes are involved in synthesis and secretion of pulmonary surfactant in pulmonary alveoli. Objective: the purpose of this study is to discuss the indication of surfactant replacement as a potential adjunctive treatment modality for SARS CoV-2, similarly treatment to neonatal respiratory distress syndrome. Methodology: we argue that SARS can be triggered by surfactant deficiency secondary to production deficiency determined by type 2 pneumocyte injuries. In this sense, we carried out a bibliographic review. Conclusion: thus, the replacement of human surfactant could be a potential treatment modality for SARS CoV-2, in the same way that it is indicated for the treatment of neonatal respiratory distress syndrome
Qualidade de vida em adultos com Transtorno de Déficit de Atenção com Hiperatividade
Authors: Caribé Cerqueira Gustavo , Pondé de Sena Eduardo
Published: February 2021

Introdução: mais de 50% das crianças com transtorno de déficit de atenção com hiperatividade (TDAH) continuam apresentando sintomas na vida adulta, com impactos no desenvolvimento psicossocial, profissional, acadêmico e emocional e, consequentemente, na qualidade de vida (QV). Objetivo: descrever características e escores da QV de dados da linha de base de uma amostra clínica de indivíduos adultos, participantes de ensaio clínico randomizado controlado do uso de estimulação transcraniana por corrente contínua no TDAH. Metodologia: sessenta indivíduos foram selecionados e submetidos à avaliação da QV com a escala de autorrelato de adultos (ASRS) e a escala da qualidade de vida em adultos com TDAH (AAQoL). Características demográficas e clínicas dos indivíduos foram descritas como frequências e porcentagens para variáveis categóricas. Resultados: a média (DP) do escore total de AAQoL foi de 43,0 (14,1). Os escores médios (DP) das subescalas foram 37,4 (15,8) para Produtividade na Vida, 38,4 (23,0) para Saúde Psicológica, 53,7 (15,5) para Perspectiva de Vida e 45,9 (22,2) para Relacionamentos. Conclusão: os resultados obtidos com o AAQoL demonstraram que os sujeitos da amostra caracterizaram o perfil do adulto portador de TDAH, onde diversas esferas da vida são comprometidas, em especial os “Relacionamentos”, em que apresentou o maior comprometimento identificado, além da “Produtividade”. Foi identificado também o consumo de bebida alcoólica e a hereditariedade com familiares também portadores, como confirma a literatura
Engels: pensador maiúsculo
Authors: Moura Mauro Castelo Branco de
Published: February 2021

Apresenta-se Engels por ele próprio em sua obra, arguto e crítico observador das condições de vida dos trabalhadores e das relações sociais ainda na juventude em Barmen/Alemanha e em Manchester na Inglaterra. Pensador maduro, vigoroso formulador das teses e análises marxistas, Engels debruça-se sobre a economia política, a concepção de história, a propriedade privada, os princípios do comunismo. Companheiro fraterno de Marx, Engels é um pensador original que em seus 200 anos de nascimento ganha a merecida atenção.
Friedrich Engels - 200 anos
Authors: Ferreira Muniz Gonçalves
Published: February 2021

Este Volume 12, N. 3 conversa sobre as contribuições de Friedrich Engels com o Professor Muniz Ferreira. O Roteiro, elaborado por Maria de Fátima Rodrigues Pereira, Pedro Leão da Costa Neto e Rodrigo Castelo explora a trajetória de estudos do entrevistado acerca da abordagem das relações internacionais em perspectiva marxista, do próprio marxismo e da obra de Marx e Engels. O resultado é uma interessante reflexão que recupera a própria trajetória intelectual de Muniz Ferreira, destacando-se aqui os estudos sobre a abordagem de Marx e Engels sobre as relações internacionais, sem perder de vista a análise da relevância das contribuições de Engels para o marxismo. Boa leitura!!

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 
Sonho de voo e medo da queda, o conflito primitivo na exploração virtuosística das técnicas aéreas circenses
Authors: Mendonça Gabriel Coelho , Gatti Daniela
Published: February 2021

O circo moderno surge em 1770 com a união dos artistas das feiras medievais e os cavaleiros militares no Astley’s Royal Amphiteatre of Arts, em Londres. Neste espaço se fundou o espetáculo de variedades circenses, baseado na valorização da potencialidade física de seus artistas, as conhecidas proezas físicas ou virtuoses acrobáticas. Nos anos de 1980, com o surgimento e a disseminação de escolas de ensino das técnicas circenses surge um movimento que se auto-intitula novo circo, ou circo contemporâneo. Em busca de postular seus cânones, os novo circenses contrapõem-se às bases do espetáculo de variedades circenses, deixando escapar termos que permitem uma compreensão reducionista acerca da potência expressiva das proezas acrobáticas. No presente trabalho, apoiando-nos nos conceitos de sonho de voo e medo da queda, propostos por Bachelard, pretendemos evidenciar a maneira pela qual os números aéreos atingem e evocam desejos primitivos e medos arquetípicos na audiência. Demonstrando, através do caso específico dos números aéreos, a ligação íntima entre as formas circenses e o conteúdo que podemos expressar dentro dessas formas, pretendemos ampliar o campo de compreensão das artes circenses em toda a sua amplitude, fortalecendo sua tradição e oferecendo ferramentas eficazes para sua renovação
Planejamento das aulas no cotidiano da alfabetização: práticas, perspectivas e desafios
Authors: Zasso Silvana Maria Bellé , Da Silva Paula Pires , Borges Daniele Simões
Published: February 2021

Neste texto apresentamos resultados de uma pesquisa realizada no ano de 2018, com professoras alfabetizadoras de um município do interior do Rio Grande do Sul, as quais participaram da formação realizada no âmbito do Pacto Nacional pela Alfabetização na Idade Certa - PNAIC 2017/2018. Problematizamos acerca da prática de planejamento das aulas pautados por referenciais que discutem o trabalho docente pelo viés coletivo e reflexivo na ação alfabetizadora e tratamos das orientações dos Cadernos de Formação do PNAIC 2012 e 2014. Explicamos a abordagem metodológica, mostrando que, por nos situarmos na área das ciências humanas e pesquisarmos fenômenos e fatos que não são possíveis de serem controlados, sustentamos a investigação numa metodologia de pesquisa qualitativa. Os dados nos permitem afirmar que há uma prática de planejamento que ora acontece de forma individual e ora coletiva e que as condições de trabalho destas profissionais nem sempre favorecem os encontros. Percebemos que as áreas de conhecimento mais abordadas por estas alfabetizadoras são a linguagem e matemática por acreditarem que no 3º ano do ciclo da alfabetização o domínio da leitura e escrita e os conceitos matemáticos precisam ser consolidados. Por fim, existe uma tendência forte de práticas de planejamento integrado.
Asplenic-hyposplenic Overwhelming Sepsis: Postsplenectomy Sepsis Revisited
Authors: Hansen Katrine , Singer Don B.
Published: July 2002

Absence of the spleen or splenic function predisposes individuals to risk of overwhelming infection. These infections are most often due to encapsulated organisms, especially pneumococcus, Haemophilus influenzae type b, and meningococcus, but any bacterial agent may cause the rapid onset of septicemia, meningitis, pneumonia, and shock characteristic of the asplenic-hyposplenic condition. The risk is greatest in infants and young children, but asplenic-hyposplenic adults also have an increased risk of infection. Prophylactic antibiotics and immunization with polyvalent pneumococcal, H. influenzae type b, and meningococcal vaccines have reduced the incidence of infections in asplenic-hyposplenic individuals, but even these measures have not eliminated the risk. Surgeons have adopted techniques to save as much splenic tissue as possible and some splenic functions, such as pitting red cells, have been preserved, but conservative surgery has not provided total protection against overwhelming infection. Therapies designed to interrupt the cascade of overwhelming sepsis have not yet been successful.

In those cases in which the spleen is surgically removed, the underlying disease or condition leading to splenectomy influences the risk of sepsis. Splenectomy incidental to other operations, such as gastrectomy, results in the lowest risk for overwhelming infection, but this is still some 35-fold greater than the risk for overwhelming infections in the general population. In increasing order of risk, the other main indications for surgical removal of the spleen are idiopathic thrombocytopenia purpura, trauma, transplantation procedures, hereditary spherocytosis, staging Hodgkin's disease, portal hypertension with hypersplenism, and thalassemia. Pathologists should comment on the risk of overwhelming sepsis when spleens are processed as surgical specimens, and should carefully weigh all splenic tissue, including accessory spleens and splenic implants (splenosis), in autopsy cases with and without overwhelming sepsis.
Caracterisation de glycolipides dans une souche de Nocardia brasiliensis
Authors: Lanélle M.A. , Asselineau J.
Published: July 2002

Several glycolipid fractions have been isolated from the bound lipids of N. brasiliensis. The main fraction was characterized as a monoester of nocardomycolic acids and D‐arabinose. Fractionation of the free lipids yielded a glycolipid fraction consisting of nocardomycouc acids, arabinose, galactose and several amino acids. This last fraction appears to be similar to wax D of Mycobacteria; mild acid treatment of this fraction produced arabinose mononorcardomycolate.
Equilibration of metabolic CO2 with preformed CO2 and bicarbonate
Authors: Hems Reginald , Saez Guillermo T.
Published: July 2002

Entry of metabolic 14CO2 into urea is shown to occur more readily than it equilibrates with the general pool of cellular plus extracellular bicarbonate plus CO2. Since the sites of CO2 production (pyruvate dehydrogenase and oxoglutarate dehydrogenase) and of fixation (carbamoylphosphate synthetase) are intramitochondrial, it is likely that the fixation of CO2 is also more rapid than its equilibration with the cytoplasmic pool of bicarbonate plus CO2. This observation may point to a more general problem concerning the interpretation of isotope data, when compartmentation or proximity of sites of production and utilisation of metabolites may result in the isotope following a preferred pathway.
Hydrodynamic studies of a DNA‐protein complex
Authors: Scheerhagen M.A. , van Amerongen H. , van Grondelle R. , Blok Joh.
Published: July 2002

Short DNA and RNA fragments complexed with the helix destabilizing protein of bacteriophage T4, GP32, have been studied in solution by electric birefringence and circular dichroism. The birefringence of the complexes is positive and the magnitude indicates that the DNA and RNA fragments become linear and rigid upon protein binding. The field free decay is biphasic. On the basis of a rigid rod approximation the slow relaxation time leads to a base‐base distance along the helix axis in the complex from 4.3 to 5.6 Å, an elongation of at least 50% compared to single‐stranded DNA.
Hydrodynamic studies of a DNA‐protein complex
Authors: Scheerhagen M.A. , Kuil M.E. , van Grondelle R. , Blok Joh.
Published: July 2002

The translational diffusion coefficient of the saturated complex of single‐stranded 145 base DNA and the helix‐destabilizing protein of phage T4, GP32, can be measured at equilibrium by means of quasi‐elastic light scattering. If the complex is considered as a rigid rod one can estimate its dimensions by combining the translational diffusion coefficient with earlier data on rotational diffusion. It was found that the average base‐base distance of the 145 base DNA in the complex is between 4.3 and 4.7 Å, while the diameter of the complex is between 44 and 68 Å. This suggests that the conformation of the complex must be such that a large amount of water is trapped.
TLC immunostaining characterization of Clostridium botulinum type A neurotoxin binding to gangliosides and free fatty acids
Authors: Takamizawa Kotaro , Iwamori Masao , Kozaki Shunji , Sakaguchi Genji , Tanaka Ryuichiro , Takayama Hiroo , Nagai Yoshitaka
Published: July 2002

TLC
GQ1b ganglioside
Anti‐neurotoxin antibody
Salt effect
Activation of yeast plasma membrane ATPase by acid pH during growth
Authors: Eraso Pilar , Gancedo Carlos
Published: July 2002

When yeast grows on media in which a final external pH lower than 4 is attained there is a 2–3‐fold increase in plasma membrane ATPase activity. This acid‐mediated activation produces a 2‐fold increase in the ATPase affinity for ATP but does not modify its optimum pH. The acid‐mediated activation is dependent on the stage of growth, only late logarithmic or stationary cells being capable of being activated by incubation in an acidic buffer. Only cells able to activate the ATPase maintain a constant internal pH when incubated in acidic buffers. It is concluded that acid‐mediated activation of the plasma membrane ATPase is a mechanism of maintaining constant internal pH during growth of yeast on acid media.
Tryptophan 54 and phenylalanine 60 are involved synergistically in the binding of E. coli SSB protein to single‐stranded polynucleotides
Authors: Casas-Finet Jose R. , Khamis Mustafa I. , Maki August H. , Chase John W.
Published: July 2002

The binding of both wild‐type and point‐mutated E. coli single‐stranded DNA‐binding (SSB) protein to poly(deoxythymidylic acid) has been studied by fluorescence and optical detection of triplet state magnetic resonance spectroscopy. Involvement of tryptophan residues 40 and 54 in stacking interactions with nucleotide bases has been inferred earlier from such studies. Investigation of a point mutation in the E. coli SSB gene product obtained by site specific oligonucleotide mutagenesis in which Phe‐60 is replaced by alanine strongly suggests the participation of Phe‐60 in the binding process, possibly by the formation of an extended stacking structure by Trp‐54, thymine and Phe‐60. This hypothesis is supported by results on the point mutations in which His‐55 is replaced by either leucine or tyrosine.
Light‐stimulated protein movement in rod photoreceptor cells of the rat retina
Authors: Philp Nancy J. , Chang Wesley , Long Kenneth
Published: July 2002

We examined the intracellular distribution of three proteins involved in the cyclic GMP cascade of visual transduction; cGMP phosphodiesterase, the α‐subunit of G‐protein and arrestin. In adult rats, light‐induced changes in the amounts of G and arrestin in the photoreceptor cell outer segments were observed both by polyacrylamide gel analysis of purified ROS and by immunocytochemical localization on retinal sections. In dark conditions, G was concentrated in the outer segments of photoreceptor cells while in the light Gα was seen in the inner segments and the outer nuclear layer. Arrestin had the opposite distribution, appearing in the inner segments and outer nuclear layer under dark conditions and in the ROS under light conditions. In contrast, PDE, the enzyme which is activated by G and inhibited by arrestin showed no light‐stimulated movement. In both light‐ and dark‐adapted retinas, PDE was localized primarily in the outer segments of the photoreceptor cells.
Effect of the bile acid taurolithocholate on cell calcium in saponin‐treated rat hepatocytes
Authors: Combettes Laurent , Dumont Micheline , Berthon Brigitte , Erlinger Serge , Claret Michel
Published: July 2002

Neomycin was used to assess the involvement of Ins (1,4,5)P3 in the Ca2+ release from the endoplasmic reticulum induced by the bile acid taurolithocholate. In saponin‐permeabilized rat hepatocytes, neomycin via its ability to bind Ins (1,4,5)P3 abolished the release of Ca2+ induced by added Ins (1,4,5)P3. In contrast, it did not alter the Ca2+ release initiated by the bile acid. In intact cells, neomycin had no effect on the [Ca2+]i rises promoted by taurolithocholate and vasopressin. It is suggested that the effect of taurolithocholate in liver is not mediated by Ins (1,4,5)P3 but results from a primary action on endoplasmic reticulum.
A time‐course study on superoxide generation and protein kinase C activation in human neutrophils
Authors: Christiansen Niels Obel
Published: July 2002

The time course of superoxide generation and membrane association of protein kinase C was studied in human neutrophils stimulated by PMA, FMLP, ionomycin and A23187. The initiation of superoxide generation in PMA; ionomycin‐ and A23187‐stimulated neutrophils was characterized by a lag period of at least 30 s in contrast to a lag period of 10–15 s in FMLP‐stimulated cells. The time course of membrane association of protein kinase C in PMA‐stimulated neutrophils was highly dependent upon the PMA concentration used for stimulation. However, membrane association of protein kinase C preceded superoxide generation in cells stimulated by 10–300 ng/ml PMA. FMLP, ionomycin and A23187 induced membrane association of protein kinase C in a few seconds and always before superoxide generation. It is concluded that membrane association of protein kinase C in PMA‐, FMLP‐, ionomycin‐ and A23187‐stimulated neutrophils precedes superoxide generation, and thereby may be part of the mechanism initiating NADPH‐oxidase activity. A simple correlation between the two parameters could not be proven, indicating that also other activation mechanisms are decisive in the activation of NADPH‐oxidase.
Sheep brain glutathione reductase: Purification and general properties
Authors: Acan N.L. , Tezcan E.F.
Published: July 2002

Sheep brain glutathione reductase was purified about 11000‐fold with an overall yield of 40%. The method included ammonium sulphate fractionation, heat denaturation, 2′,5′‐ADP Sepharose 4B and Sephadex G‐200 chromatography steps. Specific activity at the final step was 193 IU/mg. The M
r of the enzyme was found to be 116000 by gel filtration chromatography. On SDS‐PAGE, two identical subunits of M
r 64000 were obtained. From the spectral data, about 2 mol FAD per mol of enzyme were calculated.
Glycine and other amino compounds prevent chromatin precipitation at physiological ionic strength
Authors: Buche A. , Ouassaidi A. , Hacha R. , Delpire E. , Gilles R. , Houssier C.
Published: July 2002

Glycine, proline and taurine, when present in the range 0.1–0.60 M, inhibit chromatin precipitation by sodium chloride. Histone gel electrophoresis revealed that the linker histones H1 and H5 were largely depleted from the supernatant chromatin fraction at 0.2 M NaCl, while this depletion was absent in the presence of glycine. These observations are discussed in relation with the various factors which may be involved in the precipitation process.
Amino acid sequence of cinnamomin, a new member of the elicitin family, and its comparison to cryptogein and capsicein
Authors: Huet Jean-Claude , Pernollet Jean-Claude
Published: July 2002

The phytopathogenic fungi Phytophthora cinnamomi cause systemic leaf necrosis on its non‐host tobacco; in culture, it secretes a protein, called cinnamomin, which elicits leaf necrosis and protects tobacco against the pathogen Phytophthora nicotianæ, in a way similar to cryptogein and different from capsicein, elicitins of known amino acid sequences. The cinnamomin sequence has been determined and compared to other elicitins. The differences in the 3 elicitin sequences were correlated to the biological acivities: 2 lysines were ascribed as the key amino acids involved in the differential control of protection with respect to necrosis.
Amplification of the phosphorylation site ‐ ATP‐binding site cDNA fragment of the Na+,K+‐ATPase and the Ca2+‐ATPase of Drosophila melanogaster by polymerase chain reaction
Authors: Váradi András , Gilmore-Heber Maureen , Benz Edward J.
Published: July 2002

In vitro DNA‐amplification technique has been utilized to generate a 430 bp fragment of the Na+,K+‐ATPase, and a 550 bp fragment of a Ca2+‐ATPase (the sarcoplasmic reticulum‐type) of Drosophila melanogaster. The oligonucleotide primers for the DNA‐amplification (Polymerase Chain Reaction) had been designed on the basis of amino acid sequence motifs ‐ the phosphorylation site and the ATP‐binding site ‐ conserved among members of the ATPase protein family. Using the amplified cDNA‐segments as probes, we demonstrated that there is one Na+,K+‐ATPase and one Ca2+‐ATPase (sarcoplasnaic reticulum‐type) gene in the Drosophila genome. Three different mRNA species are processed from the Na+,K+‐ATPase gene and one from the Ca2+‐ATPase gene. Developmental control in expression of the Ca2+‐ATPase gene was observed.
A single amino acid difference distinguishes the human and the rat sequences of stathmin, a ubiquitous intracellular phosphoprotein associated with cell regulations
Authors: Maucuer Alexandre , Doye Valérie , Sobel André
Published: July 2002

Stathmin is a ubiquitous phosphoprotein proposed to play a general role as an intracellular relay integrating diverse regulatory signals of the cell's environment. We used a rat stathmin probe to isolate two classes of cDNAs coding for the human protein and corresponding to the usage of different polyadenylation sites. Compared to the rat sequences, they displayed a very high conservation both at the nucleic acid and the deduced protein sequence levels, with a single conservative amino acid difference. Further analysis of the protein sequence revealed novel putative phosphorylation sites, as well as internal repeated sequences which might reflect structural features involved in the molecular mechanisms by which stathmin fulfills its biological functions. The extreme conservation of the entire stathmin sequence further stresses the essential and general role of stathmin in cell regulations.
Nickel (II) as a temporary catalyst for hydroxyl radical generation
Authors: Torreilles Jean , Guérin Marie-Christine
Published: July 2002

Many in vivo studies show peroxidative damage during nickel toxicity, suggesting the generation of oxygen‐activated species. Using the murexide (5,5'‐nitrilodibarbituric acid ammonium salt) bleaching technique, we attempted to spectroscopically determine whether there are any histidyl‐peptides‐Ni(II) complexes able to catalyze a nickel‐dependent reduction of hydrogen peroxide leading to free oxygen radical production. We show that peptides containing the glycyl‐glycyl‐L‐histidyl sequence trigger nickel‐dependent production of oxygen radicals which can damage proteins, cause a rapid loss of tryptophan and a significant production of bityrosine and also induce peroxidation of polyunsaturated fatty acids. During the reaction, the histidine residue in the peptide is selectively damaged and breakdown of the peptide switches off hydroxyl‐radical production.
The 68 kDa protein of signal recognition particle contains a glycine‐rich region also found in certain RNA‐binding proteins
Authors: Herz Joachim , Flint Nicholas , Stanley Keith , Frank Rainer , Dobberstein Bernhard
Published: July 2002

Signal recognition particle (SRP) interacts with the signal sequence in nascent secretory and membrane proteins and directs them to the membrane of the endoplasmic reticulum. Membrane targeting is mediated by the 68 and the 72 kDa proteins of SRP. We have cloned and sequenced cDNA encoding the 68 kDa protein of canine signal recognition particle (SRP68). SRP68 is a basic protein comprised of 622 amino acid residues. Close to the amino terminus there is a glycine‐rich region which SRP68 has in common with some RNA‐binding proteins. SRP68 shares no detectable similarity to any of the proteins in data libraries.
Pigment stoichiometry of a newly isolated D1—D2—Cyt b559 complex from the higher plant Beta vulgaris L
Authors: Montoya G. , Yruela I. , Picorel R.
Published: July 2002

Two D1‐D2‐Cyt b
559 complexes with different pigment stoichiometry were isolated from the higher plant B. vulgaris. The procedures for isolating both complexes only differed in the washing time of the DEAE column with 50 mM Tris‐HCl, pH 7.2, 0.05% Triton X‐100 and 30 mM NaCl. When the column was washed until the eluate had an absorbance of 0.01 at 670 nm, the isolated D1—D2—Cyt b
559 complex presented a pigment stoichiometry of 6 chlorophyll a, 2 β‐carotene, and 1 cytochrome b
559 per 2 pheophytin a. In contrast, when the column was exhaustively washed until the eluate reached an absorbance of 0.005 at 670 nm, the complex had a stoichiometry of 4 chlorophyll a, 1 β‐carotene, and 1 cytochrome b
559 per 2 pheophytin a. We think that the former stoichiometry corresponds to that of the native D1—D2—Cyt b
559 complex. Moreover, both preparations showed 2 mol of pheophytin a per 1 mol of reaction center protein.
Protein aggregation and inclusion body formation in Escherichia coli rpoH mutant defective in heat shock protein induction
Authors: Gragerov A.I. , Martin E.S. , Krupenko M.A. , Kashlev M.V. , Nikiforov V.G.
Published: July 2002

Mutations in the rpoH gene, encoding σ32, an alternative factor required for transcription of the heat shock genes, result in the extensive aggregation of virtually all cellular proteins and formation of inclusion bodies both under stress and non‐stress conditions. Inhibitors of protein synthesis suppress this aggregation, suggesting that newly synthesized proteins preferentially aggregate in rpoH mutants. These data suggest that the heat shock proteins are involved in acquisition of the soluble state (i.e. correct conformation) of the bulk of intracellular proteins after their translation.
Glycoprotein‐binding site of dystrophin is confined to the cysteine‐rich domain and the first half of the carboxy‐terminal domain
Authors: Suzuki Atsushi , Yoshida Mikiharu , Yamamoto Hideko , Ozawa Eijiro
Published: July 2002

Dystrophin, a protein product of the Duchenne muscular dystrophy gene, is thought to associate with the muscle membrane by way of a glycoprotein complex which was co‐purified with dystrophin. Here, we firstly demonstrate direct biochemical evidence for association of the carboxy‐terminal region of dystrophin with the glycoprotein complex. The binding site is found to lie further inward than previously expected and confined to the cysteine‐rich domain and the first half of the carboxy‐terminal domain. Since this portion corresponds well to the region that, when missing results in severe phenotypes, our findings may provide it molecular basis of the disease.
Attraction of human monocytes by the neuropeptide secretoneurin
Authors: Reinisch Norbert , Kirchmair Rudolf , Kähler Christian M. , Hogue-Angeletti Ruth , Fischer-Colbrie Reiner , Winkler Hans , Wiedermann Christian J.
Published: July 2002

Secretoneurin is a newly discovered 33‐amino‐acid peptide derived from secretogranin II (chromogranin C) that is found in sensory afferent C‐fibers. We show here that secretoneurin triggers the selective migration of human monocytes in vitro and in vivo. Combinations of secretoneurin with the sensory neuropeptides, substance P or somatostatin, synergistically stimulate such migration. The attraction of monocytes represents the first established function of secretoneurin as a sensory neuropeptide.
Adenovirus‐mediated gene transfer to murine retinal cells in vitro and in vivo
Authors: Jomary C. , Piper T.A. , Dickson G. , Couture L.A. , Smith A.E. , Neal M.J. , Jones S.E.
Published: July 2002

Adenovirus‐mediated gene transfer to retinal cells was evaluated using the replication‐defective recombinant adenovirus vector Ad2/CMVlacZ‐1 (coding for β‐galactosidase) both in an in vitro murine culture model and in vivo in adult mice. In vitro, no difference in infectability of neuronal and glial cells was observed, and 50% of neurons expressed the exogenous gene at low viral concentration (10 pfu/cell). In vivo, intraocular injection of 3 × 106 pfu Ad2/CMVlacZ‐1 resulted in expression of the transferred β‐galactosidase gene in retinal pigment epithelium and ganglion cells. These results demonstrate that Ad2/CMVlacZ‐1 is an effective vector for gene transfer into retinal cells.
Nitric oxide donor GEA 3162 inhibits endothelial cell‐mediated oxidation of low density lipoprotein
Authors: Malo-Ranta Ulla , Ylä-Herttuala Seppo , Metsä-Ketelä Timo , Jaakkola Olli , Moilanen Eeva , Vuorinen Pauli , Nikkari Tapio
Published: July 2002

The effect of a nitric oxide (NO) donor GEA 3162 on the endothelial cell (EC)‐mediated oxidation of low density lipoprotein (LDL) was studied. In comparison to LDL incubated with EC without GEA 3162, the presence of GEA 3162 inhibited LDL oxidation by EC, as indicated by the following findings. (a) The degradation rate of LDL in macrophages was reduced to control levels. (b) The electrophoretic mobility of LDL decreased in a dose‐dependent manner, (c) The concentrations of thiobarbituric acid‐reactive substances and hydroperoxide‐derived hydroxy fatty acids were lower. (d) The breakdown of apolipoprotein B was reduced. The results indicate that GEA 3162 prevents EC‐mediated oxidation of LDL.
Structural implications for the role of the N terminus in the ‘superactivation’ of collagenases
Authors: Reinemer Peter , Grams Frank , Huber Robert , Kleine Thomas , Schnierer Susanne , Piper Michael , Tschesche Harald , Bode Wolfram
Published: July 2002

For the collagenases PMNL‐CL and FIB‐CL, the presence of the N‐terminal Phe79 correlates with an increase in proteolytic activity. We have determined the X‐ray crystal structure of the recombinant Phe79‐Gly242 catalytic domain of human neutrophil collagenase (PMNL‐CL, MMP‐8) using the recently solved model of the Met80‐Gly242 form for phasing and subsequently refined it to a final crystalographic R‐factor of 18.0% at 2.5 Å resolution. The PMNL‐CL catalytic domain is a spherical molecule with a flat active site cleft separating a smaller C‐terminal subdomain from a bigger N‐terminal domain, that harbours two zinc ions, namely a ‘structural’ and a ‘catalytic’ zinc, and two calcium ions. The N‐terminal segment prior to Pro86, which is disordered in the Met80‐Gly242 form, packs against a concave hydrophobic surface made by the C‐terminal helix. The N‐terminal Phe79 ammonium group makes a salt link with the side chain carboxylate group of the strictly conserved Asp232. Stabilization of the catalytic site might be conferred via strong hydrogen bonds made by the adjacent, likewise strictly conserved Asp233 with the characteristic ‘Met‐turn’, which forms the base of the active site residues.
Effect of glutathione depletion on formation of paramagnetic chromium in Chinese hamster V‐79 cells
Authors: Sugiyama Masayasu , Tsuzuki Katsuyuki
Published: July 2002

Incubation of Chinese hamster V‐79 cells with either buthionine sulfoximine (BSO) or diethylmaleate (DEM) prior to exposure to Na2CrO4 resulted in a depletion of GSH. ESR study shows that the depletion of GSH by BSO caused an increase in the cellular level of Cr(V) intermediate without affecting the level of Cr(III) complex, whereas the levels of Cr(V) and (III) were both suppressed by the depletion of GSH by DEM. GSH depletion by DEM decreased cellular uptake of chromate more than that by BSO. Glutathione reductase activity in cells, which is capable of reducing Cr(VI), was unaffected by BSO, however a strong inhibition was observed in cells treated with DEM, indicating that DEM affects not only GSH levels but also Cr(VI)‐reducing enzyme activity. Thus, the depletion of GSH in cells increases Cr(V) formation, based upon the effect of BSO. The role of intracellular GSH in Cr(VI) reduction is discussed.
Formation of N‐formylkynurenine suggests the involvement of apolipoprotein B‐100 centered tryptophan radicals in the initiation of LDL lipid peroxidation
Authors: Gieβauf Andreas , van Wickern Barbara , Simat Thomas , Steinhart Hans , Esterbauer Hermann
Published: July 2002

Tryptophan oxidation products were determined in pronase E digests of apo B of delipidated, native and Cu2+ oxidized LDL using a sensitive and specific HPLC method. Oxidized LDL contained N‐formylkynurenine, kynurenine and tryptamine but no oxindolylalanine and 5‐hydroxytryptophan. N‐Formylkynurenine increased from an initial value of 0.21 to 1.67 mollmol apo B within 5 h. Apo B of native LDL also contained kynurenine (0.80 mollmol) and tryptamine (0.13 mol/mol). The results support the assumption that oxidation of Trp residues in apo B is an early event and possibly an elementary reaction involved in initiating LDL oxidation.
Military pregnancies and adverse perinatal outcome
Authors: Magann E.F. , Winchester M.I. , Carter D.P. , Martin J.N. , Nolan T.E. , Morrison J.C.
Published: July 2002

Objective: To identify significant risk factors for an adverse outcome in active‐duty military women. Method: A prospective study of 105 pregnancies and their outcome. Results: The data revealed that: (1) single women more than married personnel had cesarean births when compared with forceps and vacuum (P < 0.03) or spontaneous vaginal delivery (P < 0.04); and (2) active‐duty women who gained <25 pounds during pregnancy developed preterm labor more often (P < 0.05). Conclusion: Risk factors for these adverse outcomes remain unknown.
Metal X‐ray microanalysis in the olfactory system of rainbow trout exposed to low level of copper
Authors: Julliard A. Karyn , Saucier Diane , Astic Liliane
Published: July 2002

Summary— It has recently been shown that a chronic copper exposure induces specific degeneration of olfactory receptor cells in rainbow trout; however, the exact mechanism of action of the metal is not yet known. Using X‐ray microanalysis in transmission electron microscopy, we have studied the distribution of metal in the olfactory system of fish exposed for 15, 30 and 60 days to 20 μg/l of copper. This was done in order to determine if it was accumulated in receptor cells and transported into the central nervous system via the olfactory nerve. No copper accumulation was detected either in the olfactory epithelium, in the olfactory nerve or in the olfactory bulb. The heavy metal was exclusively found within melanosomes of melanophores located in the lamina propria. After 60 days of exposure, the copper content in melanosomes was about two‐fold higher than that in controls. As far as some morphological recovery took place in the olfactory organ during the metal exposure, which lets us suppose that some detoxication mechanism occurs, it could be suggested that melanophores might be somehow involved in such a mechanism.
pH control of the plant outwardly‐rectifying potassium channel SKOR
Authors: Lacombe Benoît , Pilot Guillaume , Gaymard Frédéric , Sentenac Hervé , Thibaud Jean-Baptiste
Published: July 2002

SKOR, an Arabidopsis depolarisation‐activated K+‐selective channel, was expressed in Xenopus oocytes, and external and internal pH effects were analysed. Internal pH was manipulated by injections of alkaline or acidic solutions or by acid load from acetate‐containing medium. An internal pH decrease from 7.4 to 7.2 induced a strong (ca. 80%) voltage‐independent decrease of the macroscopic SKOR current, the macroscopic gating parameters and the single channel conductance remained unchanged. An external acidification from 7.4 to 6.4 had similar effects. It is proposed that pH changes regulate the number of channels available for activation. Sensitivity of SKOR activity to pH in the physiological range suggests that internal and external pH play a role in the regulation of K+ secretion into the xylem sap.
Increase in the adenine nucleotide translocase content of duckling subsarcolemmal mitochondria during cold acclimation
Authors: Roussel Damien , Chainier François , Rouanet Jean-Louis , Barré Hervé
Published: July 2002

Intermyofibrillar and subsarcolemmal mitochondria were isolated from duckling gastrocnemius muscle. The adenine nucleotide translocase (ANT) content of subsarcolemmal mitochondria was found to be half of that present in intermyofibrillar mitochondria. In addition, cold acclimation resulted in a 1.7‐fold increase in subsarcolemmal mitochondrial ANT content, with intermyofibrillar mitochondrial ANT remaining constant. This change in mitochondrial ANT content correlates with the previously reported cold‐induced change in the sensitivity of mitochondria to palmitate‐inhibited ATP synthesis [Roussel et al. (1998) FEBS Lett. 439, 258–262]. It is suggested that the mitochondrial ANT content enhances or reduces the fatty acid uncoupling activity in tissue, depending on the energetic state of mitochondria.
Arginine‐rich peptides are blockers of VR‐1 channels with analgesic activity
Authors: Planells-Cases R. , Aracil A. , Merino J.M. , Gallar J. , Pérez-Payá E. , Belmonte C. , González-Ros J.M. , Ferrer-Montiel A.V.
Published: July 2002

Vanilloid receptors (VRs) play a fundamental role in the transduction of peripheral tissue injury and/or inflammation responses. Molecules that antagonize VR channel activity may act as selective and potent analgesics. We report that synthetic arginine‐rich hexapeptides block heterologously expressed VR‐1 channels with submicromolar efficacy in a weak voltage‐dependent manner, consistent with a binding site located near/at the entryway of the aqueous pore. Dynorphins, natural arginine‐rich peptides, also blocked VR‐1 activity with micromolar affinity. Notably, synthetic and natural arginine‐rich peptides attenuated the ocular irritation produced by topical capsaicin application onto the eyes of experimental animals. Taken together, our results imply that arginine‐rich peptides are VR‐1 channel blockers with analgesic activity. These findings may expand the development of novel analgesics by targeting receptor sites distinct from the capsaicin binding site.
Drosophila AD3 mutation of synaptotagmin impairs calcium‐dependent self‐oligomerization activity
Authors: Fukuda Mitsunori , Kabayama Hiroyuki , Mikoshiba Katsuhiko
Published: July 2002

Genetic analysis of a Drosophila synaptotagmin (Syt) I mutant (AD3) has revealed that Tyr‐334 within the C2B domain is essential for efficient Ca2+‐dependent neurotransmitter release. However, little is known as to why a missense mutation (Tyr‐334‐Asn) disrupts the function of the C2B domain at the molecular level. Here, we present evidence that a Tyr‐312 to Asn substitution in mouse Syt II, which corresponds to the Drosophila AD3 mutation, completely impairs Ca2+‐dependent self‐oligomerization activity mediated by the C2B domain but allows partial interaction with wild‐type proteins in a Ca2+‐dependent manner. This observation is consistent with the fact that the AD3 allele is homozygous lethal but complements another mutant phenotype. We also showed that the Ca2+‐dependent C2B self‐oligomerization is inhibited by inositol 1,3,4,5‐tetrakisphosphate, a potent inhibitor of neurotransmitter release. All of these findings strongly support the idea that self‐oligomerization of Syt I or II is essential for neurotransmitter release in vivo.
GORK, a delayed outward rectifier expressed in guard cells of Arabidopsis thaliana, is a K+‐selective, K+‐sensing ion channel
Authors: Ache Peter , Becker Dirk , Ivashikina Natalya , Dietrich Petra , Roelfsema M.Rob G , Hedrich Rainer
Published: July 2002

Here we report on the molecular identification, guard cell expression and functional characterization of AtGORK, an
Arabidopsis thaliana goutward rectifying K+ channel. GORK represents a new member of the plant Shaker K+ channel superfamily. When heterologously expressed in Xenopus oocytes the gene product of GORK mediated depolarization‐activated K+ currents. In agreement with the delayed outward rectifier in intact guard cells and protoplasts thereof, GORK is activated in a voltage‐ and potassium‐dependent manner. Furthermore, the single channel conductance and regulation of GORK in response to pH changes resembles the biophysical properties of the guard cell delayed outward rectifier. Thus GORK very likely represents the molecular entity for depolarization‐induced potassium release from guard cells.
Generation and degradation of human endostatin proteins by various proteinases
Authors: Ferreras Mercedes , Felbor Ute , Lenhard Thomas , Olsen Bjorn R. , Delaissé Jean-Marie
Published: July 2002

The angiogenesis inhibitor endostatin is a fragment of the NC1 domain of collagen XVIII. The generation of endostatin has been investigated only in murine hemangioendothelioma cell cultures and was ascribed to cathepsin L. Distinct endostatin‐like fragments were detected in human tissues and serum. To identify proteinases able to generate such fragments, we incubated human NC1 with proteinases of all classes, including cathepsin L. Eleven out of 12 generate fragments with an N‐terminus within the same 15 residue stretch as those occurring physiologically, indicating that this region is sensitive to many proteinases. None correspond to mouse endostatin. However, the efficiencies of these proteinases differed markedly. Some proteinases also proved to degrade endostatin, pointing to another regulatory loop of angiogenesis.
Inhibitory serpins from rye grain with glutamine as P1 and P2 residues in the reactive center
Authors: Hejgaard Jørn
Published: July 2002

Six of seven serpins detected in grains of rye (Secale cereale) were purified and characterized. The amino acid sequence close to the blocked N‐terminus, the reactive center loop sequence and the second order association rate constant (k
a′) for irreversible complex formation with chymotrypsin were determined for each serpin. Three of four serpins containing the unusual reactive center P2–P1′ QQ↓S and one with P2–P1′ PQ↓M were equally efficient inhibitors of chymotrypsin (k
a′∼105 M−1 s−1). One serpin with P2–P1′ PY↓M was a faster inhibitor (k
a′∼106 M−1 s−1). Similar but differently organized glutamine‐rich reactive centers were recently found in grain serpins cloned from wheat [Østergaard et al. (2000) J. Biol. Chem. 275, 33272] but not from barley. The prolamin storage proteins of cereal grains contain similar sequences in their glutamine‐rich repeats. A possible adaption of hypervariable serpin reactive centers late in Triticeae cereal evolution as defence against insects feeding on cereal grains is discussed.
Aureobasidin A, an antifungal cyclic depsipeptide antibiotic, is a substrate for both human MDR1 and MDR2/P‐glycoproteins
Authors: Kino Kouichi , Taguchi Yoshitomo , Yamada Kouji , Komano Tohru , Ueda Kazumitsu
Published: July 2002

The human MDR1 gene encodes the multidrug transporter P‐glycoprotein (Pgp). Although the MDR2/Pgp shares about 80% identity at the amino acid level with the MDRI/Pgp, the MDR2/Pgp cannot act as a multidrug transporter. We examined the drug sensitivity of Saccharomyces cerevisiae expressing either the human MDR1/Pgp or MDR2/Pgp. The human MDR1/Pgp conferred about 4‐fold resistance to aureobasidin A, a cyclic depsipeptide antifungal antibiotic, on the drug‐sensitive yeast strains. Interestingly the human MDR2/Pgp also conferred about 2.5‐fold resistance to aureobasidin A. The resistance to aureobasidin A conferred by the MDR2/Pgp as well as by the MDR1/Pgp was overcome by vinblastine, verapamil, and cyclosporin A, depending on their concentrations, but not by colchicine. Aureobasidin A probably interacts directly with Pgps, because it overcame multidrug resistance of human cells and inhibited azidopine photoaffinity labeling of MDRI/Pgp in human cell membranes. These results suggest the possibility that the human MDR1 and MDR2/Pgps have conserved domain(s) for drug recognition.
Subunit a of proton ATPase F0 sector is a substrate of the FtsH protease in Escherichia coli
Authors: Akiyama Yoshinori , Kihara Akio , Ito Koreaki
Published: July 2002

Escherichia coli FtsH is a membrane‐bound ATPase with a proteolytic activity against the SecY subunit of protein translocase. We now report that subunit a of the membrane‐embedded F0 part of H+‐ATPase is another substrate of FtsH. Pulse‐chase experiments showed that subunit a is unstable when it alone (without F0 subunits b and c) was oversynthesized and that it is stabilized in the ftsH mutants. Selective and ATP‐dependent degradation of subunit a by purified FtsH protein was demonstrated in vitro. These results suggest that FtsH serves as a quality‐control mechanism to avoid potentially harmful accumulation of free subunit a in the membrane.
A cell‐density sensing factor regulates the lifetime of a chemoattractant‐induced Gα‐GTP conformation
Authors: Brazill Derrick T , Gundersen Robert , Gomer Richard H
Published: July 2002

© 1997 Federation of European Biochemical Societies.
p160ROCK, a Rho‐associated coiled‐coil forming protein kinase, works downstream of Rho and induces focal adhesions
Authors: Ishizaki Toshimasa , Naito Mamoru , Fujisawa Kazuko , Maekawa Midori , Watanabe Naoki , Saito Yuji , Narumiya Shuh
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Identification and characterization of an activated 20S proteasome in Trypanosoma brucei
Authors: To Wah-Yuen , Wang Ching C.
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Modulation of intracellular protein degradation by SSB1–SIS1 chaperon system in yeast S. cerevisiae
Authors: Ohba Masayuki
Published: July 2002

In prokaryotes, DnaK–DnaJ chaperon is involved in the protein degradation catalyzed by proteases La and ClpA/B complex as shown in E. coli. To extend this into eukaryotic cells, we examined the effects of hsp70 genes, SSA1 and SSB1, and DnaJ genes, SIS1 and YDJ1, on the growth of proteasome subunit mutants of the yeast S. cerevisiae. The results identified SSB1 and SIS1 as a pair of chaperon genes specifically involved in efficient protein turnover in the yeast, whose overexpression suppressed the growth defects caused by the proteasome mutations. Moreover, a single amino acid substitution in the putative peptide‐binding site of SSB1 protein profoundly enhanced the suppression activity, indicating that the activity is mediated by the peptide‐binding activity of this chaperon. Thus SSB1, with its partner DnaJ, SIS1, modulates the efficiency of protein turnover through its chaperon activity.