bluuebunny/crossref_metadata_2025 huggingface dataset:
Non-linear responses of net ecosystem productivity to gradient warming in a paddy field in Northeast China
Authors: Sun Yulu , Qu Fuyao , Zhu Xianjin , Sun Bei , Wang Guojiao , Yin Hong , Wan Tao , Song Xiaowen , Chen Qian
Published: June 2020

Global warming has a known impact on ecosystems but there is a lack of understanding about its impact on ecosystem processes. Net ecosystem productivity (NEP) and its components play a key part in the global carbon cycle. Analysing the impact of global warming on NEP will improve our understanding of how warming affects ecosystems. In our study, conducted in 2018, five warming treatments were manipulated (0 W, 500 W, 1000 W, 1500 W, and 3000 W) using three repetitions of far infrared open warming over a paddy field in Northeast China. NEP and its two related components, gross primary productivity (GPP) and ecosystem respiration (ER), were measured using the static chamber-infrared gas analyser method to explore the effects of different warming magnitudes on NEP. Results showed that measurement dates, warming treatments, and their interactions significantly affected NEP, ER, and GPP. Warming significantly increased NEP and its components but they showed a non-linear response to different warming magnitudes. The maximum increases in NEP and its components occurred at 1500 W warming. NEP is closely related to its components and the non-linear response of NEP may have primarily resulted from that of GPP. Gradient warming non-linearly increased GPP in the paddy field studied in Northeast China, resulting in the non-linear response of NEP. This study provides a basis for predicting the responses of carbon cycles in future climate events.
Estrategias discursivas de las instituciones públicas en la web: nuevas formas de comunicar las políticas públicas
Authors: Martínez Romagosa Maite , Zukerfeld Gilda Lucía
Published: June 2020

Este trabajo se enmarca en un proyecto UBACyT que se propone estudiar las transformaciones en los sistemas semióticos de las prácticas políticas en la Argentina pos 2001.Creemos que el uso de nuevas plataformas para la distribución de información y la interacción entre las instituciones públicas y los ciudadanos ha dado lugar a modificaciones que afectan tanto al funcionamiento de las mismas instituciones como a la constitución de la ciudadanía. Si bien estos cambios han sido estudiados desde otras ciencias sociales, no se ha focalizado en los cambios en el uso lingüístico desde una perspectiva discursiva. En particular, en este trabajo nos interesa indagar en las prácticas comunicacionales de las instituciones gubernamentales, que llamaremos, con algunos autores (Agar, 1985; Iedema, 1998; Wodak, 1997), discurso institucional.Sostenemos que el Análisis del Discurso nos permite acceder a los contenidos de las representaciones sociales (Raiter, 2016) y observar los cambios discursivos y la lucha por la acentuación de los signos (Voloshinov, 2009). Nos interesa en particular estudiar cómo los cambios en los textos y en las prácticas discursivas pueden anticipar cambios en la clasificación de sectores de la sociedad y en las prácticas sociales (Fairclough, 1992).Nuestro corpus de análisis está conformado por las páginas de inicio de dos sitios web que dependen del Gobierno de la Nación Argentina y del Gobierno de la Ciudad de Buenos Aires, respectivamente, en los que se presentan lineamientos de políticas públicas que llaman a la “participación” de la ciudadanía mediante plataformas digitales
Rodrigo Pulgar Castro. La Ética en la era de la tecnociencia. Ril Editores, Santiago de Chile, 2017. (158 pág.)
Authors: Cea Anfossi Pietro Di Angelo
Published: June 2020

..por definir
Debates en torno a los efectos de la fertilización in vitro heteróloga en la filiación
Authors: Suárez Fernández Lisandra , Pérez Orozco Luis
Published: June 2020

Los avances logrados en la Biotecnología, especialmente en el ámbito de las técnicas de reproducción humana asistida, han provocado transformaciones en la forma en que actualmente se conciben algunas instituciones jurídicas. La fertilización in vitro heteróloga de modo particular, a razón de la intervención de donantes de material genético, trastoca los fundamentos en los cuales se ha sostenido la filiación como categoría del Derecho de Familia, dando lugar a que se contemple la posibilidad de existencia de una tercera tipicidad filiatoria. Las polémicas teóricas y doctrinales que genera su implementación para los usuarios en relación a las presunciones filiatorias y la dimensión del consentimiento informado trascienden a los ordenamientos foráneos en soluciones diversas que son reflejadas de modo igualmente variado en el plano a jurisprudencial. Aún existen ordenamientos ajenos a estos cambios, generando una desprotección a los derechos de los sujetos involucrados en su utilización, cuestión que amerita una construcción normativa que atempere los basamentos teóricos de la filiación a la realidad social y la ciencia en evolución.
Los diputados chilenos y la despenalización del aborto: una aproximación desde el análisis crítico del discurso
Authors: Camacho Lucio Lina Maritza
Published: June 2020

En este artículo se analizan treinta y cuatro discursos de los diputados chilenos, durante la sesión número cincuenta, celebrada el 20 de julio de 2017, en la cual se debatió el proyecto de ley originado en mensaje que regula la despenalización de la interrupción voluntaria del embarazo en tres causales. Las intervenciones de los legisladores se entienden como un tipo de comportamiento legislativo que es prominente para observar las posiciones ideológicas sobre los intereses de las mujeres. Por medio de una escala simple que evalúa el contenido de las intervenciones se ubica a cada diputado en un eje horizontal, el cual expresa el espectro ideológico de los partidos políticos, izquierda-derecha, y el segundo del espectro discursivo, control-autonomía de las mujeres en la decisión de abortar. Los hallazgos indican que los discursos de los legisladores son coherentes con la posición ideológica del partido al que se adscriben. El artículo también demuestra que las legisladoras chilenas a la hora de representar los intereses de género como el aborto actúan según la plataforma ideológica del partido y no por las experiencias compartidas de género.
Reformulación de la justicia civil: La pertinencia de ajustarnos al debido proceso según la Corte Interamericana de Derechos Humanos
Authors: Ocaranza Vargas Giovanni
Published: June 2020

La hipótesis central de esta investigación consiste en determinar que la necesidad de la reformulación de la justicia civil, radica en adecuarse a los estándares que la Corte Interamericana de Derechos Humanos ha establecido sobre el debido proceso. Para el cumplimiento de tal meta argumentaremos que dicho resultado no puede alcanzarse por la vía de la adecuación de la actual legislación a ciertas garantías. En tal sentido, resulta necesario un cambio de paradigma entorno a los principios que informan el proceso civil en Chile. Es por este cambio en las bases sobre los cuales se erige el proceso que resulta indispensable la implementación de un nuevo Código Procesal Civil. Mediante esta obra, el lector podrá apreciar, especialmente en su parte dispositiva, que para adecuarnos a los estándares de debido proceso impuestos por la Corte IDH, será necesario una reformulación en la esencia del proceso, lo cual no puede ser alcanzado mediante parcialidades, sino más bien mediante la promulgación de un Código Procesal Civil
A (des) proteção ao trabalho da mulher: análise da normativa laboral brasileira
Authors: Kugel Lazzarin Sonilde , Kugel Lazzarin Helena
Published: June 2020

As mulheres brasileiras continuam sendo tratadas de forma desigual e discriminatória, devido a motivos históricos e culturais, bem como à falta de tratamento igualitário de gênero no ordenamento jurídico interno. O presente estudo objetiva analisar as insuficiências da legislação trabalhista – conside-rando, inclusive, as normas advindas com a “Reforma Trabalhista” –, no que diz respeito à igualdade de gênero, tendo em vista que a regulamentação do trabalho feminino demonstra falhas quanto à inserção justa e igualitária desse coletivo de mulheres no mundo do trabalho. A pesquisa é de cunho bibliográ-fico e utiliza autores nacionais e internacionais, bem como leituras e pesquisa em livros, artigos de revistas, sites oficiais e legislação nacional e internacional.
Emergencia de las mujeres indígenas en América Latina. Debates sobre género, etnicidad e identidad cultural
Authors: Fernandez Migues Sheila , Faundez Peñafiel Juan Jorge
Published: June 2020

Revisamos la situación de las mujeres indígenas en América Latina, considerando algunas de las formas de exclusión y discriminación que las afectan en el contexto de la globalización y el Estado Nación, como sistemas de dominación que se inician en la época de la conquista colonial. En particular, trabajamos en torno a las propuestas de «equidad de género», «feminismo indígena», «dualidad» y «complementariedad», en la búsqueda de alternativas para la femineidad y la interculturalidad, hacia la redefinición del Estado y la profundización democrática
Angiospermic Biodiversity of Lucknow Areas of Uttar Pradesh, India
Authors: Singh B. P. , Krishna Ajay , Singh S. C. , Kumar Sujit
Published: June 2020

The total angiospermic floral biodiversity of Lucknow district including indigenous, naturalised and cultigens comprises over 1263 plant species covering 705 genera and 140 families of which 989 species are dicotyledons and 274 species are monocotyledons. The monocotyledons are poorly represented except Poaceae and Cyperaceae. Of the 274 species of monocotyledons, 176 species belong to these two families while 98 species represent 23 different families. Poaceae is the largest family followed by Leguminosae (s.l.), Asteraceae, etc. and Euphorbia is the largest genus followed by Cassia, Cyperus, etc.
Biochemical and Microbiological Evaluation of the Effect of Processing on Cucumeropsis mannii Seed
Authors: Ileola A. O. , Omodara T. R. , Awoyinka O. A.
Published: June 2020

The determination of the effect of processing on nutritional composition and level of undigested oligosaccharide in Cucumeropsis mannii seeds was carried out. The dried melon seeds were sorted, cleaned, soaked in 60mL water for 15 min for easy removal and de-hulled to get the cotyledons, which were divided into three portions. The first portion was used unprocessed, the second portion was boiled while the third portion was fermented for analysis. The microbial load, physicochemical analyses, proximate compositions, anti-nutritional factors, antioxidant activity, vitamins, mono and oligosaccharides sugars were determined. Processing led to significant increase in the pH, moisture contents buta reduction in the total titratable acidity. Processing increased protein, fat and ash contents. Significant reduction in anti-nutritional factors of boiled and fermented melon seed was noted. Processing led to significant increase in the antioxidant activity. The saccharide sugars reduced after processing. The study observed that processing increased significantly the nutritional composition of the melon seeds but significantly reduced the anti-nutritional factors.
Magnitude of Prior Teenage Pregnancy among Women Aged 18–67 Years in Rural Southwestern Uganda
Authors: Mulogo Edgar Mugema , Ntaro Moses , Kenney Jessica , Patel Palka , Wesuta Andrew , Kawungezi Peter Chris , Bwambale Shem , Matte Michael , Bagenda Fred , Stone Geren
Published: June 2020

Objective: Teenage pregnancy is a growing health challenge among adolescents in Uganda with its magnitude varying across the different regions of Uganda. This study evaluated the magnitude and factors associated with prior teenage pregnancy among women aged 18-67 years in a rural community of Kasese district, Uganda.

Results: Fifty-two percent (52%) of the 138 women interviewed, had a prior teenage pregnancy. Having experienced a teenage pregnancy was independently associated with; occupation of current household heads (adjusted odds ratio, aOR= 0.2, 95% confidence interval, CI: 0.1 – 0.9), whether or not the current household could adequately meet their food needs (aOR= 0.1, 95% CI: 0.01 – 0.8), and whether the current household shared toilet facilities (aOR= 4.7, 95% CI: 1.0 – 21.8).

Conclusion: The findings suggest that magnitude of prior teenage pregnancy among women in this rural community is much higher than the national average. Socio economic factors at household level are contributory to prior teenage pregnancy. A multi sectoral approach integrating household livelihood improvement with health interventions targeting the girl child is proposed to curb teenage pregnancy in this context.
Characterization for Different Traits in Asiatic and European Type Carrot (Daucus carota var. sativa L.) Germplasm Lines
Authors: Singh Davinder , Dhillon Tarsem Singh , Singh Rajinder
Published: June 2020

The present investigation was carried out to characterize the eighty one carrot genotypes through variability, heritability and genetic gain for different characters. Analysis of variance revealed highly significant differences among the genotypes for all the characters. Genetic variability, heritability and genetic advance were estimated for marketable and its component yield traits in 81 lines of carrot (Daucus carota var. sativa L.). The study showed high ranges for total yield (4.9-9.3 kg plot-1), marketable yield (4.6-9.2 kg plot-1), root length (13.8-30.8 cm) and root weight (101.4-127.2 g). Genotypes PC-161, PC-15, PC-173 (red), PCO-30, PCO-5, PCO-7 (Orange) PCP-2, PCP-1 and PCP-17B (Purple) were superior with respect to total and marketable yield per plot along with various horticultural traits. Magnitude of phenotypic coefficient of variation (PCV) was higher than corresponding genotypic coefficient of variation (GCV) for all the characters which indicated the role of environment on the character expression. Quantitative traits like Anthocyanin content (mg 100 g-1), Total sugar content (%), Carotene content (mg 100g-1) and Dry matter content (%) have higher values of GCV, heritability, genetic advance and genetic gain hence these were the most important traits for applying selection in carrot for crop improvement.
Hydro-geologic, Corrosivity and Geotechnical Implications of Geoelectric Sounding Survey at FUTA Cooperative Housing Estate, Ilaramokin near Akure Southwestern Nigeria
Authors: Adeyemo I. A. , Iwuala V. O. , Fadare T. S. , Otayemi E. O.
Published: June 2020

Aim: Groundwater potential evaluation, corrosivity and geotechnical competence investigations of the subsurface layers were carried at FUTA Cooperative Housing Estate, Ilaramokin near Akure Southwestern Nigeria in order to facilitate proper positioning of facilities within the estate.

Methodology: A total of 14 vertical electrical sounding (VES) data was acquired across the estate using Schlumberger array.

Results: The VES survey results delineated 3 - 5 geoelectric layers across the estate which corresponds to the topsoil, weathered layer, partially weathered basement, partially fractured basement and the presumed fresh bedrock. The groundwater potential map of the estate revealed that the groundwater potential is moderate (GWP of 0.5 - 0.75) to high (GWP of 0.75 - 1.0) in the western and southeastern parts of the estate. The depth slice corrosivity maps (1.0, 2.0 and 3.0 m) of the estate indicated that the corrosivity of the subsurface layers within the estate reduces with depth. The 0.5 m depth slice isoresistivity map of the estate indicates that the central and a segment of the southwestern parts of the estate are characterized with low resistivity (50 - 100 Ωm) indicating incompetence, while the estate flanks (west and east) are considered to be moderately competent (100 - 350 Ωm). Likewise, the 1.0 m depth slice isoresistivity map of the estate reveals that the central and western parts of the estate are characterized with low resistivity (50 - 100 Ωm) suggesting incompetent layer, while the northern, southern and eastern parts of the estate are moderately competent (100 - 350 Ωm).

Conclusion: These results suggested that structural failures can be expected on any engineering structures sited at the central, northern, southern and eastern parts of the estate. Groundwater efforts should be concentrated at the western and southeastern parts of the estate.
Transferred NOE analyses of conformations of peptides as bound to membrane bilayer of phospholipid; mastoparan‐X
Authors: Wakamatsu Kaori , Higashijima Tsutomu , Fujino Masahiko , Nakajima Terumi , Miyazawa Tatsuo
Published: July 2002

The 270‐MHz 1H‐NMR spectra of mastoparan‐X, a tetradecapeptide toxin from hornet venom, have been observed in the presence of vesicles of perdeuterated dilauroylphosphatidylcholine ([2H64]DLPC). By the analysis of transferred nuclear Overhauser effects (TRNOE), the mastoparan‐X molecule as bound to [2H64]DLPC vesicles is found to take an α‐helical form in the C‐terminal (and central) part. The TRNOE analysis with perdeuterated phospholipid is a unique method for the elucidation of conformation of peptide (and drug) molecules as bound to phospholipid membranes.
Purification and properties of suppressor seryl‐tRNA:ATP phosphotransferase from bovine liver
Authors: Mizutani Takaharu , Hashimoto Atsushi
Published: July 2002

Seryl‐tRNASer
CmCA:ATP phosphotransferase was purified 1200‐fold from bovine liver by ultracentrifugation at 150 000 × g, chromatography on DEAE—cellulose, fractional precipitation with ammonium sulfate, chromatography on hydroxyapatite, gel filtration on Sephacryl S‐300 and affinity chromatography on Blue Sepharose. Molecular mass was estimated as 135–145 kDa. The K
m values for ATP and ser‐tRNASer
CmCA were 2 mM and 21 nM, respectively. This enzyme did not react with ser‐tRNASer
IGA, tyr‐tRNA or thr‐tRNA.
Evidence for cytochrome bc1 complex involvement in nitrite reduction in a photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans
Authors: Urata Katsuro , Satoh Toshio
Published: July 2002

The cytochrome bc
1 complex which has a quinol : cytochrome c
2 oxidoreductase activity was isolated from a photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans. When duroquinol (tetramethyl‐p‐benzoquinol) was used as the electron donor, maximum nitrite reduction activity was obtained in an anaerobic mixture containing the isolated cytochrome bc
1 complex, nitrite reductase and cytochrome c
2 Nitrite reduction in chromatophore membranes, as well as in the reconstituted system, was inhibited in the presence of antimycin A. No nitrite reduction occurred in the absence of cytochrome c
2. These results indicated that the cytochrome bc
1 complex involved in photosynthetic electron transfer also donates reducing equivalents to the dissimilatory nitrite reductase via cytochrome c
2 in this photodenitrifier.
Sialosyllactotetraosylceramide, a novel ganglioside antigen detected in human carcinomas by a monoclonal antibody
Authors: Nilsson Olle , Månsson Jan-Eric , Lindholm Leif , Holmgren Jan , Svennerholm Lars
Published: July 2002

A novel ganglioside was detected in a small cell lung carcinoma by TLC‐immunostaining of gangliosides with a monoclonal antibody, the C‐50 MAb. Structural characterization showed this ganglioside to be IV3NeuAc‐LcOse4Cer, a hitherto unknown ganglioside. This ganglioside has also been detected as a minor component in many different carcinomas using the C‐50 MAb. The normally dominant CA‐50 ganglioside antigen is IV3NeuAc. III4Fuc‐LcOse4Cer. Based upon solid‐phase binding to IV3NeuAc, III4‐LcOse4Cer and IV3NeuAc‐LcOse4Cer it is concluded that the C‐50 MAb recognizes an epitope present in sialylated type I carbohydrate chains.
C‐terminal lysine residues of fibrinogen fragments essential for binding to plasminogen
Authors: Christensen Ulla
Published: July 2002

C‐terminal lysine residue
Fibrinogen
Plasminogen
Lysine binding site
Fibrinogen fragment D
Fibrinogen fragment E
Castanospermine: a potent inhibitor of sucrase from the human enterocyte‐like cell line Caco‐2
Authors: Trugnan Germain , Rousset Monique , Zweibaum Alain
Published: July 2002

The addition of castanospermine (5–50 μM) to a culture medium of Caco‐2 cells results in a specific suppression of sucrase activity without modification of the biosynthesis of the enzyme. This effect is due to a direct inhibiting effect of castanospermine on Caco‐2 sucrase activity. This inhibition is time‐dependent (half‐maximum efficiency at 10 min for 100 nM), enhanced by preincubation (suggesting a strong interaction with the enzyme), dose‐dependent (ED50 at 4 nM after 1 h preincubation period) and of the fully non‐competitive type. The calculated K
i (2.6 nM) suggests that castanospermine is the most potent inhibitor of sucrase so far reported.
The activity of γ‐glutamyltranspeptidase in regenerating rat liver
Authors: Sulakhe Susan J.
Published: July 2002

γ‐Glutamyltranspeptidase is expressed at low levels in the liver of the male Fischer 344 rat where it exhibits 15‐fold purification and 33% recovery in isolated plasma membranes. While the activity of the enzyme is unaltered in regenerating liver 24 h after partial hepatectomy, it increases steadily thereafter over a period of one week. Seven days after partial hepatectomy the enzyme is maximally activated: 5.6‐fold in liver homogenates and 5.3‐fold in isolated liver plasma membranes. The enzyme declines in activity over the next fourteen days and is expressed at normal levels three weeks after partial hepatectomy. These results demonstrate that the activity of γ‐glutamyltranspeptidase increases in regenerating liver but that the increase is out of phase with the proliferative response.
Nucleotide sequence of the open reading frames adjacent to the coat protein cistron in potato virus X genome
Authors: Morozov S.Yu. , Lukasheva L.I. , Chernov B.K. , Skryabin K.G. , Atabekov J.G.
Published: July 2002

The cloned cDNA copies corresponding to 1300 nucleotides adjacent to the 3'‐terminal poly(A) tract of the potato virus X (PVX) genome have been sequenced. The amino acid sequences of three open reading frames were deduced from the nucleotide sequence. Two putative small nonstructural polypeptides corresponding to the open reading frames adjacent to the coat protein cistron possess some properties of membrane‐associated proteins. Direct sequence homology and common structural peculiarities exist between the PVX small proteins and the putative small nonstructural proteins encoded by RNA 2 of hordeiviruses and furoviruses
Nucleotide sequence analysis of cDNAs encoding a bovine galanin precursor protein in the adrenal medulla and chemical isolation of bovine gut galanin
Authors: Rökaeus Åke , Carlquist Mats
Published: July 2002

A description is given of the primary structure of a bovine adrenal medullary precursor protein (123 amino acids), containing a nonrepetitive galanin sequence, different in four amino acid positions from pig galanin, structural information deduced from the nucleotide sequence analysis of two cDNA clones. Pairs of lysine and arginine residues separate the galanin sequence N‐terminally from the initiating methionine and a 29 amino acid leader peptide and C‐terminally from a 59 amino acid peptide. The deduced amino acid composition was confirmed by the isolation and determination of the amino acid composition for bovine intestinal galanin. Northern blot analysis revealed the presence of an apparent single galanin mRNA species, of approx. 850–900 bases in size, which was present in a population of chromaffin cells scattered throughout the bovine adrenal medulla.
Gene cloning from yeast chromosome‐specific mini‐library Isolation of the SRP1 ‐related DNA sequence located on chromosome XV
Authors: Marguet D. , Fantino E. , Lauquin G.J.-M.
Published: July 2002

We have isolated and purified chromosome XV DNA molecules from the yeast S. cerevisiae using contour‐clamped homogeneous electric field (CHEF) gel electrophoresis. A chromosome‐specific mini‐library was constructed and the element of the SRP1 (serine‐rich protein) related sequence family located within chromosome XV was isolated by in situ colony hybridization with an SRP1 probe. Results indicated that (i) a single‐copy sequence homologous to SRP1 is present within chromosome XV; (ii) this sequence lies within the 2.3 kb HindIII fragment of the plasmid pXVAc6; (iii) the lack of a specific transcript from this SRP1 ‐related element suggests that it could be considered as a pseudogene.
Growth hormone‐albumin conjugates Reduced renal toxicity and altered plasma clearance
Authors: Poznansky Mark J. , Halford Jennifer , Taylor Donna
Published: July 2002

The effective therapeutic use of many small peptides such as growth hormone has been limited by their small molecular masses and rapid clearance by the kidneys. Moreover, various degrees of nephrotoxicity have been reported for small proteins which are readily filtered at the level of the glomerulus. We have attempted to circumvent this drawback by conjugating growth hormone (somatotropin) to serum albumin in an effort to alter the peptide's pharmacokinetics while retaining its biological activity.
Characterization of the structural genes for the DNA‐binding protein H‐NS in Enterobacteriaceae
Authors: La Teana A. , Falconi M. , Scarlato V. , Lammi M. , Pon C.L.
Published: July 2002

The promoter region of Escherichia coli hns, the structural gene for the DNA‐binding protein H‐NS, has been identified by use of a promoter search vector and the in vivo transcriptional start point by primer extension analysis. The homologous hns genes of two other Enterobacteriaceae, Proteus vulgaris and Serratia marcescens, were identified by heterologous hybridization with a DNA probe derived from E. coli hns, cloned and sequenced. Taking into account only the invariant nucleotides and amino acids, the homology of H‐NS among the three organisms was found to be ⪢ 70% at the DNA level and ⪢ 75% at the protein level. The three hns genes were also found to have nearly identical transcriptional and translational signals.
Cupredoxines of obligate methylotroph
Authors: Dinarieva Tatiana , Netrusov Alexander
Published: July 2002

Cells of type I obligate methylotroph Methylobacillus flagellatum KT, which can use only methanol or methylamine as carbon and energy sources, possess two types of blue copper proteins: amicyanin and azurin. During exponential phase of growth amicyanin was the only blue copper‐containing protein in the cells from methylamine medium. Azurin‐type proteins appeared in these cells when they reached the stationary growth phase. In contrast, amicyamin could not be detected in methylamine‐grown cells when they are from stationary growth phase. Cells from methanol‐containing medium possess only azurin, amicyanin could not be detected in all growth phases. Kinetic studies with methylamine dehydrogenase and amicyanin purified from M. flagellatum KT cells show that amicyanin may serve as a primary electron acceptor from methylamine and methylamine dehydrogenase in this type I of obligate methylotroph.
Substrate isomerization inhibits ribulosebisphospate carboxylase‐oxygenase during catalysis
Authors: Edmondson Daryl L. , Kane Heather J. , Andrews T.John
Published: July 2002

The inhibition of purified spinach ribulosebisphosphate carboxylase‐oxygenase which occurs progressively during catalysis in vitro is caused by accumulation of at least two tight‐binding inhibitors at the catalytic site. Reduction of these inhibitors with NaB3H4, followed by dephosphorylation, produced a mixture of xylitol and arabinitol, thus identifying one of them as D‐xylulose 1,5‐bisphosphate. It was formed during carboxylation, presumably by a stereochemically incorrect reprotonation of the 2,3‐enediolate intermediate bound at the catalytic site. Under the conditions used, this epimerization occurred approximately once for every 400 carboxylation turnovers. Another inhibitor may be 3‐keto‐D‐arabinitol 1,5‐bisphosphate which would also be formed by misprotonation of the enediolate intermediate, but at C‐2 rather than at C‐3.
Thrombopoietic activity of human interleukin‐6
Authors: Nagasawa Toshiro , Orita Tetsuro , Matsushita Jun-ichi , Tsuchiya Masayuki , Neichi Tomohiro , Imazeki Ikuo , Imai Nobuo , Ochi Norimichi , Kanma Hiroshi , Abe Tsukasa
Published: July 2002

Thrombopoietin (TPO), a regulatory factor in platelet production, was purified from the conditioned medium of TNK‐01 cells cultured in the presence of human interleukin‐1. The N‐terminal sequence of purified TPO was determined to be VPPGEDSKDVAAPHRQPLT, identical to that of the N‐terminal region of human interleukin‐6 (IL‐6). Two forms of TPO with molecular masses of 24 and 27 kDa were identified as IL‐6 by Western analysis using an anti‐IL‐6 antibody. Commercial recombinant human IL‐6 produced in Escherichia coli, stimulated megakaryocyte colony formation in the presence of mouse interleukin‐3 and increased the number of peripheral platelets in mice in a dose‐dependent manner. From these results, it is concluded that human IL‐6 has thrombopoietic activity.
Inhibition of digestive proteinase of stored grain coleoptera by oryzacystatin, a cysteine proteinase inhibitor from rice seed
Authors: Liang C. , Brookhart G. , Feng G.H. , Reeck G.R. , Kramer K.J.
Published: July 2002

Electrophoresis of midgut extracts from the rice weevil, Sithopilus oryzae, and the red flour beetle, Tribolium castaneum, in polyacrylamide gels containing sodium dodecyl sulfate and gelatin revealed there was one major proteinase (apparent molecular mass = 40 000) in the rice weevil and two major proteinase (apparent molecular masses = 20 000 and 17 000) in the red flour beetle. The pH optima using [3PH]casein as substrate were about pH 6.8. for the rice weevil and pH 5.2 for the red flour beetle. Use of specific inhibitors, including L‐trans‐epoxysuccinyl‐leucylamino‐(4‐guanidino)‐butane (E‐64). p‐chloromercuriphenylsulfonic acid (PCMS), and oryzacystatin, indicated that nearly all of the proteinase activity against casein was contributed by cysteine proteinases. The estimated IC10 values for oryzacystatin were 2 × 10−6 M and 4 × 10−7 M when tested against midgut extracts from T. castaneum and S. oryzae, respectively.
The three‐dimensional structure of the bifunctional proteinase k/α‐amylase inhibitor from wheat (PKI3) at 2.5 Å resolution
Authors: Zemke Klaus J. , Müller-Fahrnow Anke , Jany Klaus-Dieter , Pal Gour P. , Saenger Wolfram
Published: July 2002

Wheat germ contains an inhibitor for proicinase K, called PK13 (M
1 ∼ 19600) which simultaneously inhibits α‐amylase. PK13 was crystallized, space group P21, α = 43.02 (5) Å, n = 65.18 (7) Å, c = 32.33 (4) Å, β = 112.79 (9), X‐ray data were collected to 2.5 Å resolution, the structure solved by molecular replacement on the basis of the atomic coordinates of the homologous Erythrina caffra DE‐3 inhibitor, and refined with simulated annealing techniques with a current R‐factor of 21%. The three‐dimensional structure of PK13 is stabilized by two disulfide bridges and has a central β‐barrel with distorted β‐structure. In analogy to related inhibitors, the binding site for proteinase K is assumed to be located on the surface of the protein (amino acids residues 66–67), although the 75–76 peptide bond is cleaved upon binding.
Staurosporine induces a neuronal phenotype in SH‐SY5Y human neuroblastoma cells that resembles that induced by the phorbol ester 12‐O‐tetradecanoyl phorbol‐13 acetate TPA)
Authors: Jalava Annika , Heikkilä Jari , Lintunen Minnamaija , Åkerman Karl , Pahlman Sven
Published: July 2002

Treatment of SH‐SY5Y human neuroblastoma cells with the protein kinase inhibitor staurosporine, induced both morphological and functional differentiation in these cells. The effects of staurosporine were comparable to those induced by the protein kinase C (PKC) activator. 12‐O tetradecanoyl phorbol 13‐acetate (TPA), with respect to induction of neuronal differentiation, i.e. neurite outgrowth, inhibition of DNA synthesis, induction and down‐regulation of c‐myc protein expression, induction of mRNA for both neuropeptide Y (NPY) and growth associated protein 43 (GAP‐43) and stimulation of tyrosine hydroxylase expression. Staurosporine failed to translocate PKC to the membrane fraction or to stimulate phosphorylation of the endogenous PKC substrate M
r 80,000 (p8O). Instead, staurosporine inhibited TPA‐induced phosphorylation of p80.
In vivo phosphorylation of the 30‐kDa protein of tobacco mosaic virus
Authors: Watanabe Yuichiro , Ogawa Toshiya , Okada Yoshimi
Published: July 2002

The 30‐kDa protein of tobacco mosaic virus, which is involved in cell‐to‐cell movement function, is phosphorylated in tobacco protoplasts. To investigate which portion of the protein is phosphorylated we inoculated several truncated 30‐kDa protein mutants into protoplasts and determined whether or not those truncated proteins are phosphorylated. The results showed that amino acid residues 234–261 of the 30‐kDa protein are required for this phosphorylation.
Inhibition of the degradation of the precursor and of the mature β1 integrin subunit by different protein synthesis inhibitors and by ATP depletion
Authors: Vekeman Sylvie , Jaspers Martine , Cassiman Jean-Jacques
Published: July 2002

A stabilization of both the precursor and the mature β1 integrin subunit was observed in metabolically labeled human skin fibroblasts and Molt‐4 T lymphocytes upon addition of protein synthesis inhibitors or by ATP depletion. Differentia] effects of protein synthesis inhibitors are reported since the slow degradation of the mature β1 subunit was sensitive to cycloheximide but not to puromycin. We also show that the half‐life of the mature subunit was not dependent on intracellular lysosomal degradation or on ubiquitination suggesting that VLA turn‐over occurs at the cell surface and might involve proteins or proteases with short half‐life.
Human insulin gene enhancer‐binding proteins in pancreatic α and β cell lines
Authors: Clark Andrew R. , Petersen Helle V. , Read Martin L. , Scott Valerie , Michelsen Birgitte K. , Docherty Kevin
Published: July 2002

Electrophoretic mobility shift assays were performed using oligonucleotides corresponding to known protein binding sites within the human insulin gene enhancer and nuclear extracts from mouse pancreatic α and β cell lines. The results demonstrate that a previously described factor, IUF‐1, binds to three sites at −82 (the CT1 box), −215 (the CT2 box), and −319 (the CT3 box) in the human insulin gene enhancer. IUF‐1 was present only in β but not in α cells, while all other DNA‐binding proteins were present in both cell lines. IUF‐1 may therefore be an important determinant of insulin gene β cell‐specific expression.
The secondary structure of the ferredoxin transit sequence is modulated by its interaction with negatively charged lipids
Authors: Horniak Ladislav , Pilon Marinus , van't Hof Ron , de Kruijff Ben
Published: July 2002

Import of proteins into chloroplasts depends on an N‐terminal transit sequence. Transit sequences contain little primary sequence similarity and therefore recognition of these sequences is thought to involve specific folding. To assess the conformational flexibility of the transit sequence, we studied the transit peptide of preferredoxin (trfd) by circular dichroism. In buffer, trfd is in a random coil conformation. A large increase in α‐helix was induced in the presence of micelles or vesicles formed by anionic lipids. Less pronounced changes in secondary structure were induced by zwitterionic detergents but no changes were observed in the presence of neutral detergents or vesicles composed of phosphatidylcholine.
The amino acid sequence previously attributed to a protein kinase or a TCP1‐related molecular chaperone and co‐purified with phytochrome is a β‐glucosidase
Authors: Gus-Mayer Sabine , Brunner Harald , Schneider-Poetsch Hansjörg A.W. , Lottspeich Friedrich , Eckerskorn Christoph , Grimm Rudolf , Rüdiger Wolfhart
Published: July 2002

A 60 kDa protein (P60) co‐purified with phytochrome was identified as avenacosidase, a β‐glucosidase which is part of the defense system of Avena
sativa. An antiserum raised against P60 was used to isolate a cDNA clone coding for the complete amino acid sequence of P60. The cDNA‐derived amino acid sequence contained the partial sequences described before for a protein kinase [(1989) Planta 178, 199–206] and for a TCP1‐related molecular chaperone [(1993) Nature 363, 644–647] co‐purified with phytochrome. We conclude that these activities were related to minor contaminants and that only sequences of avenacosidase had been obtained.
Formation of SRP‐like particle induces a conformational change in E. coli 4.5S RNA
Authors: Lentzen Georg , Dobberstein Bernhard , Wintermeyer Wolfgang
Published: July 2002

E. coli P48 protein is homologous to the SRP54 component of the eukaryotic signal recognition particle. In vivo, P48 is associated with 4.5S RNA which shares a homology with eukaryotic SRP RNA. To study the interaction between P48 and 4.5S RNA in vitro, we used 4.5S RNA with fluorescein coupled to the 3′‐terminal ribose. Upon binding of P48, the fluorescent 4.5S RNA shows a substantial decrease in fluorescence. Fluorescence quenching as well as anisotropy measurements reveal that the effect is not due to a direct interaction of P48 with the dye. This suggests that the binding of P48 induces a conformational change in 4.5S RNA which affects the structure at the 3′ end of the RNA. From equilibrium titrations with fluorescent 4.5S RNA, a dissociation constant of O.15 μm is obtained for the RNA · protein complex. The formation of the complex is not affected by GTP binding to or hydrolysis by P48.
A stably folded presecretory protein associates with and upon unfolding translocates across the membrane of mammalian microsomes
Authors: Schlenstedt Gabriel , Zimmermann Maria , Zimmermann Richard
Published: July 2002

The presecretory protein ppcecDHFR, a hybrid between preprocecropin A and dihydrofolate reductase, is transported into mammalian microsomes post‐translationally, i.e. independently of ribosome and signal recognition particle. Upon staging the transport process, stably folded ppcecDHFR bound to mammalian microsomes and subsequently translocated across the membrane. Membrane association depended on the signal peptide but involved neither ATP nor an N‐ethylmaleimide‐sensitive microsomal protein. Membrane insertion of bound ppcecDHFR did not necessitate unfolding of the DHFR domain but depended on ATP and an N‐ethylmaleimide‐sensitive microsomal protein. Completion of translocation relied on unfolding of the DHFR domain. Thus mammalian microsomes have the capability of transporting a bound and folded precursor protein, i.e. to trigger unfolding of a precursor protein on the membrane surface.
Prothymosin α receptors on peripheral blood mononuclear cells
Authors: Cordero Oscar J. , Sarandeses Conceptión , Nogueira Montserrat
Published: July 2002

125I‐Labeled prothymosin α (ProTα) was used to study the presence and characteristics of receptors for ProTα on human peripheral blood mononuclear cells (PBMC). The kinetics of 125I‐ProTα binding to PBMC was fast at 37°C, whilst it required 50 min to reach equilibrium at 4°C and room temperature. Analysis of steady state binding data by the method of Scatchard and by unlabeled ProTα competition experiments identified two binding sites with an apparent equilibrium dissociation constant of 216–321 pM for the high‐affinity receptor and of 11.4–21.1 nM for the low‐affinity one; the sites per cell ranged from 1,479 to 1,519 and from 47,547 to 56,169, respectively. The kinetically derived equilibrium dissociation constant agreed with these data and showed no interaction between receptors.
A new function of Nm23/NDP kinase as a differentiation inhibitory factor, which does not require it's kinase activity
Authors: Okabe-Kado Junko , Kasukabe Takashi , Hozumi Motoo , Honma Yoshio , Kimura Narimichi , Baba Hideo , Urano Takeshi , Shiku Hiroshi
Published: July 2002

We recently identified a differentiation inhibiting factor (I‐factor) in mouse myeloid leukemia M1 cells as a murine homolog of nm23‐H2/nucleoside diphosphate kinase (NDPK)‐B gene product. We examined the I‐factor activities of several authentic nm23/NDPK proteins, i.e recombinant rat NDPK α and β, recombinant mouse nm23‐M1 and ‐M2, and recombinant human nm23‐H1 and ‐H2 containing a mutant nm23‐H2His protein lacing NDPK activity. Almost all these nm23/NDPK proteins showed I‐factor activity. Moreover, to understand the active domain exhibiting I‐factor activity of nm23‐H2 protein lacking NDPK activity, we have investigated the I‐factor activities of some truncated nm23‐H2 proteins. The truncated nm23‐H2 protein containing N‐terminal peptide 1–60 retained the I‐factor activity. These results provide the first evidence for a function of nm23/NDPK as a differentiation inhibiting factor in leukemic cells, that is independent of its NDPK activity and dependent on the presence of N‐terminal peptide.
The secG deletion mutation of Escherichia coli is suppressed by expression of a novel regulatory gene of Bacillus subtilis
Authors: Kontinen Vesa P. , Helander Ilkka M. , Tokuda Hajime
Published: July 2002

SecG, a membrane component of E. coli protein translocase, stimulates the translocation of proteins across the cell membrane through the cycle of topology inversion, which is coupled to the membrane‐insertion and deinsertion cycle of SecA [Nishiyama et al. (1996) Cell 85, 71–81]. A gene of B. subtilis able to suppress the cold‐sensitive phenotype of the secG deletion mutant of E. coli was cloned and found to encode a novel regulatory protein, ScgR. Similarity search revealed homology with known proteins such as GlnR of B. subtilis. Plasmid‐encoded ScgR stimulated protein translocation in the deletion mutant. ScgR increased the proportion of cardiolipin at the expense of phosphatidylglycerol, but did not affect the composition of other lipid components of the cell, suggesting that the increased cardiolipin level compensates for the SecG function and thereby stimulates protein translocation.
Quality of life and insomnia validation study for a specifically designed questionnaire
Authors: Jambon B , Le Gal M , Pilate C
Published: July 2002

A questionnaire was devised to assess changes in the quality of life of insomniac patients when treated by a hypnotic. The questionnaire was validated in two groups of patients (60 non-insomniac and 52 insomniac patients). The results of this study confirmed the relevance of the questionnaire and its power to distinguish between the two groups of patients. A second study carried out simultaneously in 58 insomniac patients receiving treatment showed the internal reliability of the questionnaire and its reproducibility after 3 to 5 days.
Regulation of the Golgi structure by the α subunits of heterotrimeric G proteins
Authors: Yamaguchi Tomohiro , Nagahama Masami , Itoh Hiroshi , Hatsuzawa Kiyotaka , Tani Katsuko , Tagaya Mitsuo
Published: July 2002

Disassembly of the Golgi apparatus is elicited by the action of nordihydroguaiaretic acid (NDGA) and this disassembly is prevented by the activation of heterotrimeric G proteins. In the present study we showed that overexpression of Gαz or Gαi2 significantly suppresses the disassembly of the Golgi apparatus induced by NDGA. Overexpression of Gβ1γ2, on the other hand, had no effect on NDGA‐induced Golgi disassembly. Gαz neither blocked Golgi disassembly induced by brefeldin A or nocodazole, nor interfered with protein transport, suggesting its specificity on the action of NDGA. Our results suggest that the α subunits of heterotrimeric G proteins are responsible for the maintenance of the Golgi structure.
The Arabidopsis thaliana cDNA sequencing projects1
Authors: Delseny Michel , Cooke Richard , Raynal Monique , Grellet Françoise
Published: July 2002

© 1997 Federation of European Biochemical Societies
Bromocriptine is a strong inhibitor of brain nitric oxide synthase: possible consequences for the origin of its therapeutic effects
Authors: Renodon A. , Boucher J.-L. , Sari M.-A. , Delaforge M. , Ouazzani J. , Mansuy D.
Published: July 2002

The ergot alkaloid bromocriptine (BKT) was found to act as a strong inhibitor of purified neuronal nitric oxide synthase (NOS) (IC50=10±2 μM) whereas it was poorly active towards inducible macrophage NOS (IC50>100 μM). BKT affects the activation of NOS by calmodulin, as it not only inhibits l‐arginine oxidation to NO and l‐citrulline but also NADPH oxidation and calmodulin‐dependent cytochrome c reduction catalyzed by neuronal NOS. These results suggest that BKT could exert some of its therapeutic effects by interfering with the NOS‐dependent formation of nitric oxide and/or superoxide ion in various tissues.
Cloning and expression of superoxide dismutase from Aquifex pyrophilus, a hyperthermophilic bacterium
Authors: Lim Jae-Hwan , Yu Yeon Gyu , Choi In-Geol , Ryu Jae-Ryeon , Ahn Byung-Yoon , Kim Sung-Hou , Han Ye Sun
Published: July 2002

A superoxide dismutase (SOD) gene of Aquifex pyrophilus, a marine hyperthermophilic bacterium, was cloned, sequenced, expressed in Escherichia coli, and its gene product characterized. This is the first SOD from a hyperthermophilic bacterium that has been cloned. It is an iron‐containing homo‐oligomeric protein with a monomeric molecular mass of 24.2 kDa. The DNA‐derived amino acid sequence is more similar to those of known Mn‐ and Fe‐SODs from thermophilic archaea than of Cu, Zn‐SODs. The metal binding residues found in all SOD sequences from different species are also conserved in A. pyrophilus SOD. The protein is biochemically active only as an oligomer and is resistant to thermal denaturation.
The 14‐3‐3 protein binds its target proteins with a common site located towards the C‐terminus
Authors: Ichimura Tohru , Ito Mitsuki , Itagaki Chiharu , Takahashi Masuhiro , Horigome Tsuneyosi , Omata Saburo , Ohno Shigeo , Isobe Toshiaki
Published: July 2002

The 14‐3‐3 protein family binds a variety of proteins in cell‐signaling pathways, but the structural elements necessary for the ligand binding are poorly understood. Here we demonstrate that the `box‐1' region, which spans residues 171–213 in the η‐isoform and was previously identified as the binding site of 14‐3‐3 to the phosphorylated tryptophan hydroxylase, plays a critical role in the interaction with many target proteins. Using a series of truncated 14‐3‐3 mutants, we show that the mutant 167–213 carrying box‐1 binds bacurovirus‐expressed Raf‐1 and Bcr protein kinases to the similar extent as the full‐length 14‐3‐3 in a phosphorylation‐dependent manner, while the mutants lacking this region abolish the binding activity. Furthermore, the box‐1 region also appears essential for binding of 14‐3‐3 to more than 40 phosphoproteins found in the brainstem extract. These results suggest that the box‐1 region, consisting of helices 7 and 8 in the tertiary structure, is a common structural element whereby the 14‐3‐3 protein binds many, if not all, target proteins.
E. coli translation initiation factor IF2 – an extremely conserved protein. Comparative sequence analysis of the infB gene in clinical isolates of E. coli
Authors: de A. Steffensen Søren A , Poulsen Ane B , Mortensen Kim K , Sperling-Petersen Hans U
Published: July 2002

The functionally uncharacterised N‐terminal of translation initiation factor IF2 has been found to be extremely variable when comparing different bacterial species. In order to study the intraspecies variability of IF2 the 2670 basepairs nucleotide sequence of the infB gene (encoding IF2) was determined in 10 clinical isolates of E. coli. The N‐terminal domains (I, II and III) were completely conserved indicating a specific function of this region of IF2. Only one polymorphic position was found in the deduced 890 amino acid sequence. This Gln/Gly490 is located within the central GTP/GDP‐binding domain IV of IF2. The results are further evidence that IF2 from E. coli has reached a highly defined level of structural and functional development.