bluuebunny/crossref_metadata_2025 huggingface dataset:
An Examination of the Self-Evaluations Over Time of the 3.11 Tsunami Survivors Regarding Their Post-Disaster Recovery
Authors: LEE Young-Jun , Hanada Shinichi
Published: August 2020

In this study, the factors that affected the sense of recovery of the victims of the Great East Japan Earthquake were analyzed from both short-term and long-term perspectives. We used the results of questionnaire surveys conducted in 2013 and 2017 amongst residents of Noda Village, Iwate Prefecture, to determine the factors that affected the sense of recovery of their personal lives as well as their sense of the recovery of the village itself. We also examined how differences in the choice of temporary housing affected their sense of recovery. Utilizing decision tree analysis, we clarified differences in the key factors that impacted their short-term and long-term perspectives as well as the sense of their own recovery and their sense of the village’s recovery. Because the introduction of publicly-funded rental housing deviated from the temporary housing networks available in previous disasters, there remains the possibility that although the short-term sense of recovery of those in publicly-funded temporary housing was relatively low, it seemed to increase over the long-term.
CURATORSHIP PRAXISES TOWARDS ‘DISPLACED ASSETS’ IN POST-1989 MUSEUM PROJECTS: TWO EXHIBITIONS AT WARSAW’S XAWERY DUNIKOWSKI MUSEUM
Authors: Toniak Ewa
Published: August 2020

Two exhibitions at the Xawery Dunikowski Museum of Sculpture at the Królikarnia Palace, branch of the National Museum in Warsaw: the ‘Inventorying’ Display-Research Project, which was a kind of a public inventory of the sculpture collection (2012) and the Exhibition ‘The Estate. Sculptures from the collection of the Von Rose family and films and photographs from the archive of Zofia Chomętowska’ (2015) are case studies serving the Author to analyse curatorship practices with respect to the collections whose major part is composed of ‘displaced assets’, first of all from the so-called ‘Regained Territories’. In the words of the Chief Curator at the Królikarnia Museum since 2011 and the Exhibitions’ Curator Agnieszka Tarasiuk: it is a troublesome collection testifying to a difficult heritage and not yielding to conservation. The paper’s methodological basis is the museum exhibits’ provenance research conducted by R. Olkowski, L.M. Kamińska, and M. Romanowska-Zadrożna, while its context is found in the programme assumptions of the Strategy for the Operations and Development of the National Museum in Warsaw 2010–2020 worked out by the former National Museum’s Director Piotr Piotrowski. One of its priorities is to clarify the origins of the collections of unknown provenance, and settling accounts with their former owners. Furthermore, the question related to constructing museum’s genealogy and the memory of history of the period immediately following WWII in the new socio-political situation in Poland after 1989 is posed. The position for dealing with collections’ provenance research introduced by P. Piotrowski was liquidated following the Director’s dismissal in 2012. The paper forms part of a bigger whole.
Geragogy as a pedagogical subdyscipline
Authors: Dubas Elżbieta
Published: August 2020

The text presents geragogy ( first of all from from polish research perspective) that is frequently described as pedagogy of old age, but here presented as a pedagogical subdiscipline dealing with education aimed at old age. It points to the beginnings of geragogy and its position in the system of scientific research concerning old age and ageing that are, on the one hand pedagogical and on the other remaining in the realm of gerontology. It presents the subject matter of geragogy highlighting the key research context that education plays here, including its various functions. The work outlines the sections and contemporary fields of geragogical research and also reveals the character of geragogy as science as well as its methodology. Additionally, it presents the model of a geragogue as a specialist dealing with educational support for people (not only the elderly ones) as far as experiencing old age and the processes of ageing are concerned.
Projeto Matematicando: um caminho interdisciplinar
Authors: Sant’Ana Nádia Aparecida dos Santos , Oliveira Marilza Maria de
Published: August 2020

Este artigo visa apresentar o Projeto “Matematicando” realizado com alunos do Ensino Fundamental de uma escola pública do interior do Estado de Minas Gerais. O referido projeto é anual, consistindo na realização de diversas atividades interdisciplinares durante uma semana, com a participação ativa do corpo discente e docente da instituição. O objetivo central é motivar os alunos a realizarem pesquisas relativas a alguns conteúdos matemáticos. O mês que antecede a realização das atividades é dedicado à elaboração e produção do material que será utilizado. Há também a colaboração da comunidade externa à escola. Nas suas diversas edições, o Matematicando se revela como um espaço de grande interação entre professores e alunos, promovendo a ampliação e consolidação dos conhecimentos. A cada ano, verifica-se um maior envolvimento e expectativa dos estudantes. Os resultados obtidos indicam que o projeto propicia que os alunos tenham diferentes olhares para os conteúdos escolares, relacionando os saberes matemáticos com outras áreas do conhecimento de forma mais consistente e objetiva.
Cortical RORβ is required for layer 4 transcriptional identity and barrel integrity
Authors: Clark Erin A , Rutlin Michael , Capano Lucia S , Aviles Samuel , Saadon Jordan R , Taneja Praveen , Zhang Qiyu , Bullis James B , Lauer Timothy , Myers Emma , Schulmann Anton , Forrest Douglas , Nelson Sacha B
Published: August 2020

Retinoic acid-related orphan receptor beta (RORβ) is a transcription factor (TF) and marker of layer 4 (L4) neurons, which are distinctive both in transcriptional identity and the ability to form aggregates such as barrels in rodent somatosensory cortex. However, the relationship between transcriptional identity and L4 cytoarchitecture is largely unknown. We find RORβ is required in the cortex for L4 aggregation into barrels and thalamocortical afferent (TCA) segregation. Interestingly, barrel organization also degrades with age in wildtype mice. Loss of RORβ delays excitatory input and disrupts gene expression and chromatin accessibility, with down-regulation of L4 and up-regulation of L5 genes, suggesting a disruption in cellular specification. Expression and binding site accessibility change for many other TFs, including closure of neurodevelopmental TF binding sites and increased expression and binding capacity of activity-regulated TFs. Lastly, a putative target of RORβ, Thsd7a, is down-regulated without RORβ, and Thsd7a knock-out alone disrupts TCA organization in adult barrels.
A broad mutational target explains a fast rate of phenotypic evolution
Authors: Besnard Fabrice , Picao-Osorio Joao , Dubois Clément , Félix Marie-Anne
Published: August 2020

The rapid evolution of a trait in a clade of organisms can be explained by the sustained action of natural selection or by a high mutational variance, that is the propensity to change under spontaneous mutation. The causes for a high mutational variance are still elusive. In some cases, fast evolution depends on the high mutation rate of one or few loci with short tandem repeats. Here, we report on the fastest evolving cell fate among vulva precursor cells inCaenorhabditisnematodes, that of P3.p. We identify and validate causal mutations underlying P3.p's high mutational variance. We find that these positions do not present any characteristics of a high mutation rate, are scattered across the genome and the corresponding genes belong to distinct biological pathways. Our data indicate that a broad mutational target size is the cause of the high mutational variance and of the corresponding fast phenotypic evolutionary rate.
Orderly assembly underpinning built-in asymmetry in the yeast centrosome duplication cycle requires cyclin-dependent kinase
Authors: Geymonat Marco , Peng Qiuran , Guo Zhiang , Yu Zulin , Unruh Jay R , Jaspersen Sue L , Segal Marisa
Published: August 2020

Asymmetric astral microtubule organization drives the polarized orientation of the S. cerevisiae mitotic spindle and primes the invariant inheritance of the old spindle pole body (SPB, the yeast centrosome) by the bud. This model has anticipated analogous centrosome asymmetries featured in self-renewing stem cell divisions. We previously implicated Spc72, the cytoplasmic receptor for the gamma-tubulin nucleation complex, as the most upstream determinant linking SPB age, functional asymmetry and fate. Here we used structured illumination microscopy and biochemical analysis to explore the asymmetric landscape of nucleation sites inherently built into the spindle pathway and under the control of cyclin-dependent kinase (CDK). We show that CDK enforces Spc72 asymmetric docking by phosphorylating Nud1/centriolin. Furthermore, CDK-imposed order in the construction of the new SPB promotes the correct balance of nucleation sites between the nuclear and cytoplasmic faces of the SPB. Together these contributions by CDK inherently link correct SPB morphogenesis, age and fate.
The oldest record of the Steller sea lionEumetopias jubatus(Schreber, 1776) from the early Pleistocene of the North Pacific
Authors: Tsuzuku Nahoko , Kohno Naoki
Published: August 2020

The extant genera of fur seals and sea lions of the family Otariidae (Carnivora: Pinnipedia) are thought to have emerged in the Pliocene or the early Pleistocene in the North Pacific. Among them, the Steller sea lion (Eumetopias jubatus) is the largest and distributed both in the western and eastern North Pacific. In contrast to the limited distribution of the current population around the Japanese Islands that is now only along the coast of Hokkaido, their fossil records have been known from the middle and late Pleistocene of Honshu Island. One such important fossil specimen has been recorded from the upper lower Pleistocene Omma Formation (ca. 1.36–0.83 Ma) in Kanazawa, Ishikawa Prefecture, Japan, which now bears the institutional number GKZ-N 00001. Because GKZ-N 00001 is the earliest fossil having been identified as a species of the sea lion genusEumetopias, it is of importance to elucidate the evolutionary history of that genus. The morphometric comparisons were made among 51 mandibles of fur seals and sea lions with GKZ-N 00001. As results of bivariate analyses and PCA based on 39 measurements for external morphologies with internal structures by CT scan data, there is almost no difference between GKZ-N 00001 and extant male individuals ofE. jubatus. In this regard, GKZ-N 00001 is identified specifically as the Steller sea lionE. jubatus. Consequently, it is recognized as the oldest Steller sea lion in the North Pacific. About 0.8 Ma, the distribution of the Steller sea lion had been already established at least in the Japan Sea side of the western North Pacific.
The influence of bait on remote underwater video observations in shallow-water coastal environments associated with the North-Eastern Atlantic
Authors: Jones Robyn E. , Griffin Ross A. , Januchowski-Hartley Stephanie R. , Unsworth Richard K.F.
Published: August 2020

The use of baited remote underwater video (BRUV) for examining and monitoring marine biodiversity in temperate marine environments is rapidly growing, however many aspects of their effectiveness relies on assumptions based on studies from the Southern Hemisphere. The addition of bait to underwater camera systems acts as a stimulus for attracting individuals towards the camera field of view, however knowledge of the effectiveness of different bait types in northern temperate climbs is limited, particularly in dynamic coastal environments. Studies in the Southern Hemisphere indicate that oily baits are most effective whilst bait volume and weight do not impact BRUV effectiveness to any great degree. The present study assesses the influence of four bait types (mackerel, squid, crab and no bait (control)) on the relative abundance, taxonomic diversity and faunal assemblage composition at two independent locations within the North-Eastern Atlantic region; Swansea Bay, UK and Ria Formosa Lagoon, Portugal. Two different bait quantities (50 g and 350 g) were further trialled in Swansea Bay. Overall, patterns showed that baited deployments recorded statistically higher values of relative abundance and taxonomic diversity when compared to un-baited deployments in Swansea Bay but not in Ria Formosa Lagoon. No statistical evidence singled out one bait type as best performing for attracting higher abundances and taxonomic diversity in both locations. Faunal assemblage composition was however found to differ with bait type in Swansea Bay, with mackerel and squid attracting higher abundances of scavenging species compared to the crab and control treatments. With the exception of squid, bait quantity had minimal influence on bait attractiveness. It is recommended for consistency that a minimum of 50 g of cheap, oily fish such as mackerel is used as bait for BRUV deployments in shallow dynamic coastal environments in the North-Eastern Atlantic Region.
A Study of MHD Casson Fluid Flow over a Permeable Stretching Sheet with Heat and Mass Transfer
Authors: Asogwa K. K. , Ibe A. A.
Published: August 2020

In this paper, we study the numerical approach of MHD Casson fluid flow over a permeable stretching sheet with heat and mass transfer taking into cognizance the various parameters present. A set of suitable local similarity transformations are used to non-dimensionalize the governing equations of the present problem. The system of ordinary differential equations are solved numerically by MATLAB bvp4c solver. The effect of the involved parameters on Velocity, Temperature, and Concentration, Skin friction coefficient, Nusselt number and Sherwood number has been studied and numerical results are presented graphically and in tabular form. The numerical results are in good agreement with those of the results previously published in the literature.
The chloroplast ATP synthetase consist of the subunits α, β, γ, δ, ϵ and proteolipid only
Authors: Süss Karl-Heinz , Manteuffel Renate
Published: July 2002

The subunit stoichiometry of the ATP synthetase (CF1‐CF0) immunoprecipitated from Triton X‐100 extracts of chloroplast thylakoid membranes was determined to be α3, β3, γ, δ, ϵ (CF1) and I0.3, II0.6–0.9, III4(6) (CF0). Antibodies against the polypeptides α, β, γ, δ, I, II and ϵ combined specifically with the isolated subunits as analysed by the protein blotting method. Applying this technique, antibodies against the CF1 subunits were found to form complexes with the corresponding polypeptides of thylakoids, whereas those against I (M
r 20 000) and II (M
r 17 000) combined with M
r 26 000 and M
r 24 500 membrane polypeptides, respectively. The M
r 26 000 polypeptide was identified as the major subunits of the light‐harvesting chlorophyll a/b‐protein (LHCP) complex and the M
r 24 500 component seems to be functionally connected with this complex. From the results it is concluded that the chloroplast ATP synthetase consists of the subunit of the α, β, γ, δ, ϵ and III (proteolipid only and that proteolytically altered LHCP polypeptides bind artifically to the protein complex during isolation.
An iron‐activated alcohol dehydrogenase
Authors: Scopes R.K.
Published: July 2002

An alcohol dehydrogenase isolated from Zymomonas mobilis was found to be activated by ferrous ions but not by zinc, after inactivation with metal‐complexing agents. Cobaltous ions also re‐activated to a lesser extent. It is suggested that in this species the alcohol dehydrogenase naturally contains iron. Kinetic studies on the iron‐treated enzyme indicate an ‘alcohol activation’ phenomenon, which may have physiological relevance in overcoming product inhibition during fermentation.
Chlorophyll a/b proteins of Photosystem I
Authors: Lam Eric , Ortiz William , Malkin Richard
Published: July 2002

The chlorophyll a/b‐protein complex (LHCPI) associated with Photosystem I (PS I) has been isolated from spinach thylakoids and further fractionated into two chlorophyll‐containing complexes by sucrose gradient centrifugation. The lighter fraction contains two polypeptides with relative molecular masses of 23 and 22 kDa and has been designated as LHCPIa. The denser fraction is enriched in a 20 kDa polypeptide and has been named LHCPIb. Both fractions have a chlorophyll a/b ratio of 3.5 ± 0.5. The absorption spectra and 77 K fluorescence emission spectra of the fractions show distinct characteristics with LHCPIb having a fluorescence maximum at 730 nm at 77 K while LHCPIa shows a maximum at 680 nm. The optical activities of the chlorophyll a/b complexes and the antenna‐depleted PS I have been examined by circular dichroism (CD) in the near‐UV and visible regions of the spectrum. All the LHCPI complexes show strong CD signals at 648, 485 and 340 nm which are absent in the antenna‐depleted PS I complex.
Correlation of photochemical cycle, H+ release and uptake, and electric events in bacteriorhodopsin
Authors: Drachev L.A. , Kaulen A.D. , Skulachev V.P.
Published: July 2002

The kinetics of 3 photoinduced responses of bacteriorhodopsin have been compared: spectral changes, pH shifts in a suspension of open purple membrane sheets and electric potential generation by the sheets incorporated into a lipid‐impregnated collodion film. In the presence of a pH‐buffer, the H+ release by bacteriorhodops in was shown to correlate with the formation of the M412 intermediate and the microsecond phase of the potential generation. The H+/M412 ratio is equal to 0.7±0.1 if the ionic strength of the solution is high. In the absence of the buffer, the H+ release proved to be much slower than spectral and electric responses. The kinetics of H+ uptake by bacteriorhodopsin is close to M412 decay and to the electrogeneous millisecond phase in both the presence and absence of the pH buffer. The bacteriorhodopsin‐induced proton release phase accounts for about 20%, and the uptake phase for about 80% of the overall potential. This is compatible with the model assuming that the proton start‐out point ‐ possibly, the protonated Schiff base connecting lysine 216 with retinal ‐ is closer to the outer rather than the inner (cytoplasmic) surface of the bacterial membrane.
Human glucagon‐like peptides 1 and 2 activate rat brain adenylate cyclase
Authors: Hoosein Naseema M. , Gurd Ruth S.
Published: July 2002

Two human glucagon‐like peptides, GLP‐1 and GLP‐2, which are coencoded with pancreatic glucagon in the preproglucagon gene, do not significantly inhibit [125I]monoiodoglucagon binding to rat liver and brain membranes and do not activate adenylate cyclase in liver plasma membranes. Nevertheless, GLP‐1 and GLP‐2 were each found to be potent stimulators of both rat hypothalamic and pituitary adenylate cyclase. Only 30–50 pM concentrations of each peptide elicited half‐maximal adenylate cyclase stimulation. Our data suggest that GLP‐1 and GLP‐2 may be neurotransmitters and/or neuroendocrine effectors, which would account for their high degree of sequence conservation through vertebrate evolution.
Steady‐state kinetics of F1‐ATPase
Authors: Roveri Oscar A. , Calcaterra Nora B.
Published: July 2002

The kinetic behaviour of the ATPase activity of beef heart F1 depends largely on the exposure of the enzyme to some anionic ligands such as sulphate and/or EDTA. F1 prepared in the presence of such anions exhibited a triphasic kinetic pattern whereas F1 from which those anions were removed by dialysis exhibited only two K
m values for ATP. Conversely to what has been previously reported, bicarbonate did not linearize F2‐ATPase kinetics. Moreover, anion activation cannot be simply explained by promotion of ADP release but mainly by an increase in affinity of the third catalytic site for ATP.
Spatial structures in mitochondrial suspension induced by cation efflux
Authors: Holmuhamedov E.L. , Evtodienko Yu.V.
Published: July 2002

The formation of spatial structures in a thin unstirred layer of a mitochondrial suspension has been studied. It is shown that the structure formation depends on the state of the ion‐transporting systems of mitochondria and that pattern development coincides with the activation of cation efflux from preloaded mitochondria. Spatial structure formation is an energy‐dependent process and is suppressed by respiratory chain inhibitors. Patterning is also inhibited by EGTA, EDTA and ruthenium red, reflecting the requirement for divalent cation translocation in mitochondria for the studied phenomenon.
Iron promoters of the Fenton reaction and lipid peroxidation can be released from haemoglobin by peroxides
Authors: Gutteridge John M.C.
Published: July 2002

Hydroxyl radical
Lipid peroxidation
Transferrin iron binding
Heme‐protein iron release
Hemoglobin Fenton catalyst
Desferrioxamine
Bleomycin‐detectable iron
The phosphorylation at Thr 124 of simian virus 40 large T antigen is crucial for its oligomerization
Authors: Montenarh Mathias , Müller Dorothee
Published: July 2002

SV40 large T antigen is phosphorylated at up to ten different amino acids clustered in an N‐terminal and a C‐terminal part of the polypeptide chain. The N‐terminal phosphorylated residues include Ser 123 and Thr 124. We have analyzed the oligomerization, the complex formation with the cellular oncoprotein p53 and the DNA‐binding properties of T antigen from two different SV40 transformed cell lines which have either an amino acid exchange at Ser 123 to Phe (W7) or Thr 124 to Ile (D29). In comparison to wild‐type T antigen both mutant T antigens have a slightly reduced binding affinity for both binding sites, I and II, of SV40 DNA. Phosphorylation at both residues of T antigen is not essential for formation of the complex with p53. Only the phosphorylation at Thr 124 seems to be critical for the formation of high molecular mass oligomers. Our data support the hypothesis that the oligomerization of T antigen seems to be implicated in viral DNA replication.
Identification of three pertussis toxin substrates (41, 40 and 39 kDa proteins) in mammalian brain Comparison of predicted amino acid sequences from G‐protein α‐subunit genes and cDNAs with partial amino acid sequences from purified proteins
Authors: Itoh Hiroshi , Katada Toshiaki , Ui Michio , Kawasaki Hiroshi , Suzuki Koichi , Kaziro Yoshito
Published: July 2002

We have determined the partial amino acid sequences of the 40 kDa protein, one of the three pertussis toxin substrates in porcine brain. Purified 40 kDa protein from porcine brain was completely digested with TPCK‐trypsin. Digested peptides were separated by reverse‐phase HPLC and subjected to analysis by gas‐phase protein sequencing. Several sequences of porcine brain 40 kDa protein completely matched with those which were deduced from the nucleotide sequences of the human Gi2α gene and rat Gi2α cDNA. On the other hand, the previously determined sequences of the rat brain 41 and 39 kDa proteins were in complete agreement with the predicted amino acid sequences of rat Gi1α and Goα cDNAs, respectively.
Lignin peroxidase compound III
Authors: Wariishi Hiroyuki , Gold Michael H.
Published: July 2002

At pH 3.0 in the absence of a reducing substrate and the presence of only 20 equivalents of H2O2, lignin peroxidase (LiP) is readily converted to LiP compound III (LiPIII) (Fe(III)O−
2 ‐ complex). LiPIII which is produced via the reaction of LiP compound II (LiPII) with H2O2 is not part of the peroxidase catalytic cycle, and is readily and irreversibly inactivated. Veratryl alcohol (VA), a Phanerochaete chrysosporium secondary metabolite, protects the enzyme from inactivation via two mechanisms. Acting as a substrate, VA reduces LiPII to regenerate the native enzyme. Secondly, the binding of VA to LiPIII rapidly displaces O.−
2/HO.−
2, thereby converting LiPIII directly to the native enzyme. VA is not consumed during this displacement reaction. These results help to explain the role of VA in stabilizing the enzyme in the presence of excess H2O2.
A fifth chloroplast‐encoded polypeptide is present in the photosystem II reaction centre complex
Authors: Webber A.N. , Packman L. , Chapman D.J. , Barber J. , Gray J.C.
Published: July 2002

The small polypeptides present in the pea photosystem II reaction centre complex have been separated by high‐resolution SDS‐polyacrylamide gel electrophoresis and characterised by N‐terminal amino acid sequencing. Two polypeptides are identified as the α‐ and β‐subunits of cytochrome b559 which are the products of the pdbE and psbF genes. A third polypeptide, of 4.5 kDA, is identified as the product of a small open reading frame in chloroplast DNA. The gene for this polypeptide , psbI, is located just downstream of the psbK gene chloroplast DNA.
Identification of a procarboxypeptidase A‐truncated protease E binary complex in human pancreatic juice
Authors: Moulard Maxime , Kerfelec Brigitte , Mallet Bernard , Chapus Catherine
Published: July 2002

The characterization, in human pancreatic juice, of a binary complex associating procarboxypeptidase A with a 32 kDa inactive glycoprotein (G32) is reported in this paper. Free G32 was isolated after dissociation of the binary complex. N‐terminal sequence analysis revealed a complete homology between this protein and human protease E (HPE 1), except for the two strongly hydrophobic N‐terminal residues (Val‐Val) which are missing in G32. This protein might be a truncated protease E highly analogous to the subunit III of the ruminant procarboxypeptidase A‐S6 ternary complex. The analogy with bovine subunit III is further supported by interspecies reassociation experiments showing that bovine procarboxypeptidase A can specifically bind human G32.
Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii share features with both mitochondrial and higher plant chloroplast presequences
Authors: Franzén Lars-Gunnar , Rochaix Jean-David , von Heijne Gunnar
Published: July 2002

Chloroplast transit peptides from the green alga Chlamydomonas reinhardtii have been analyzed and compared with chloroplast transit peptides from higher plants and mitochondrial targeting peptides from yeast, Neurospora and higher eukaryotes. In terms of length and amino acid composition, chloroplast transit peptides from C. reinhardtii are more similar to mitochondrial targetting peptides than to chloroplast transit peptides from higher plants. They also contain the potential amphiphilic α‐helix characteristic of mitochondrial presequences. However, in similarity with chloroplast transit peptides from higher plants, they contain a C‐terminal region with the potential to form an amphiphilic β‐strand. As in higher plants, transit peptides that route proteins to the thylakoid lumen consist of an N‐tenninal domain similar to stroma‐targeting transit peptides attached to a C‐terminal apolar domain that share many characteristics with secretory signal peptides.
Solubilization of the bombesin receptor from Swiss 3T3 cell membranes
Authors: Coffer Arnold , Fabregat Isabel , Sinnett-Smith James , Rozengurt Enrique
Published: July 2002

Bombesin and structurally related peptides including gastrin releasing peptide (GRP) are potent mitogens for Swiss 3T3 cells. Here we attempted to solubilize bombesin receptors under conditions in which the ligand (125I‐labelled GRP) was prebound to the receptor prior to detergent extraction. We found that 125I‐GRP‐receptor complexes were solubilized from Swiss 3T3 cell membranes by using the detergents taurodeoxycholate or deoxycholate. These detergents promoted ligand‐receptor solubilization in a dose‐dependent manner. In contrast, a variety of other detergents including Triton X‐100, octylglycoside, CHAPS, digitonin, cholic acid and n‐dodecyl‐β‐D‐maltoside, were much less effective. Addition of guanosine 5'‐[γ‐thio]triphosphate (GTPγS) to ligand‐receptor complexes isolated by gel nitration enhanced the rate of ligand dissociation in a concentration‐dependent and nucleotide‐specific manner. Our results demonstrate for the first time the successful solubilization of 125 I‐GRP‐receptor complexes from Swiss 3T3 cell membranes and provide evidence for the physical association between the ligand‐receptor complex and a guanine nucleotide binding protein(s).
Effect of ADP/ATP antiporter conformational state on the suppression of the nonspecific permeability of the inner mitochondrial membrane by cyclosporine A
Authors: Novgorodov Sergey A. , Gudz Tatjana I. , Kushnareva Yulia E. , Zorov Dmitry B. , Kudrjashov Yury B.
Published: July 2002

The influence of the conformational state of ADP/ATP antiporter on the efficiency of the inhibitory effect of cyclosporine A on the Ca2+‐induced nonspecific permeability of the inner mitochondrial membrane has been studied. Carboxyatractiloside, the inhibitor of ADP/ATP‐antiporter, was shown to prevent the cyclosporine A‐induced suppression of the nonspecific permeability. The Carboxyatractiloside enect was displayed only in mitochondria depleted of adenine nucleotides. Bifunctional SH reagent, phenylarsine oxide, was also able to reverse the effect of cyclosporine A. The data are consistent with the suggestion that cyclosporine A causes suppression of the nonspecific permeability due to its effect on the ADP/ATP antiporter conformation.
The complete amino acid sequence of bullfrog (Rana catesbeiana) parvalbumin pI4.97
Authors: Sasaki Takuji , Tanokura Masaru , Asaoka Kazuo
Published: July 2002

The primary structure of the parvalbumin (pI4.97) from bullfrog, Rana catesbeiana, skeletal muscle has been determined. It is composed of 110 amino acid residues and a free amino terminus, and has a molecular mass of 11919. The amino add sequences which are thought to be functionally important sites are also conserved in the bullfrog parvalbumin. The calculated phylogenic tree indicates that this parvalbumin belongs to the α group of parvalbumins. The mutation rate of parvalbumin was fairly rapid in frogs compared to mammals. The subdivergence of frogs is also discussed.
Ethanol has an acute effect on bile acid biosynthesis in man
Authors: Axelson Magnus , Mörk Birgitta , Sjövall Jan
Published: July 2002

A single dose of ethanol, 0.4 g/kg body weight, was found to give a 5–15 fold increase of the plasma concentrations of 7α‐hydroxy‐cholesterol and 7α‐hydroxy‐4‐cholesten‐3‐one in humans. The rise was maximal 4 h after ethanol ingestion, was dose‐dependent and was not seen in a cholecystectomized subject. The effect was selective for these and some other 7α‐hydroxylated C27‐intermediates in bile acid bioxynthesis. The changes are compatible with an acute stimulation of cholesterol 7α‐hydroxylase possibly due to an ethanol‐induced inhibition of gallbladder contraction resulting in an interruption of the enterohepatic circulation of bile acids. The effect is of interest in relation to the influence of ethanol consumption on cardiovascular and gallstone diseases.
Lipoprotein lipases and vitellogenins in relation to the known three‐dimensional structure of pancreatic lipase
Authors: Persson Bengt , Jörnvall Hans , Olivecrona Thomas , Bengtsson-Olivecrona Gunilla
Published: July 2002

A 106‐residue region of high similarity between lipoprotein/pancreatic/hepatic lipases and Drosophila vitellogenins encompasses four β‐strands with all residues but one strictly conserved or conservatively replaced between the structures, and enclosing the putative active site Ser‐152. The properties suggest a common folding pattern but the region probably does not function as an ‘interface recognition site’ in the lipases, although it might well bind fatty acid esters of ecdysteroids or single lipid molecules in the vitellogenins. C‐terminally of this 106‐residue region, a surface loop (‘flap’) covers the active site. No residue within this loop is conserved through all lipases, but adjacent segments exhibit 60–70% residue identity, Hepatic and lipoprotein lipases probably hydrolyze both soluble and emulsified substrates at the same site. They lack residues corresponding to a second active site postulated in pancreatic lipase to account for hydrolysis of soluble substrates. In addition, due to structural differences the flap could prevent entry of soluble substrate molecules into the active site of pancreatic lipase.
Molecular cloning and nucleotide sequence determination of the regulator region of mecA gene in methicillin‐resistant Staphylococcus aureus (MRSA)
Authors: Hiramatsu Keiichi , Asada Kazumi , Suzuki Eiko , Okonogi Kenji , Yokota Takeshi
Published: July 2002

Molecular cloning and nucleotide sequence analysis were performed for the identification or the regulator genes of methicillin resistance in the genome of a MRSA strain N315. Two open reading frames (orfs) were identified in the 5′‐flanking region of the mecA gene. Predicted amino acid sequences of these orfs showed extensive homology to the co‐inducer and the repressor protein of the penicillinase (PCase) production in Staphylococcus aureus as well as in Bacillus licheniformis. These orfs are considered to encode putative co‐inducer and repressor proteins specific for the regulation of methicillin resistance in MRSA.
Calcium signaling in lymphocytes and ELF fields Evidence for an electric field metric and a site of interaction involving the calcium ion channel
Authors: Liburdy Robert P.
Published: July 2002

Calcium influx increased during mitogen‐activated signal transduction in thymic lymphocytes exposed to a 22 mT, 60 Hz magnetic field (E
induced=1.7 mV/cm. 37°C, 60 min). To distinguish between an electric or a magnetic field dependence a special multi‐ring annular cell culture plate based on Faraday's Law of Induction was employed. Studies show a dependence on the strength of the induced electric field at constant magnetic flux density. Moreover, exposure to a pure 60 Hz electric field or to a magnetically‐induced electric field of identical strength resulted in similar changes in calcium transport. The first real‐time monitoring of [Ca2+]
i
during application of a 60 Hz electric field revealed an increase in [Ca2+]
i
observed 100 s after mitogen stimulation; this suggests that the plateau phase rather than the early phase of calcium signalling was influenced. The hypothesis was tested by separating, in time, the early release of calcium from intracellular stores from the influx of extracellular calcium. In calcium‐free buffer, 60 Hz fields exerted little influence on the early release of calcium from intracellular stores. In contrast, addition of extracellular calcium during exposure enhanced calcium influx through the plasma membrane. Alteration of the plateau phase of calcium signalling implicates the calcium channel as a site of field interaction. In addition, an electric field exposure metric is mechanistically consistent with a cell‐surface interaction site.
The expression of lactase enzymatic activity and mRNA in human fetal jejunum Effect of organ culture and of treatment with hydrocortisone
Authors: Villa Marco , Ménard Daniel , Semenza Giorgio , Mantei Ned
Published: July 2002

Very sensitive procedures were developed for the parallel determination of intestinal lactase (LPH) activity and the cognate mRNA. Between 14 and 20 weeks of gestation, lactase activity is low and varies only slightly; at 37 weeks, a relatively high level of activity is observed. The amounts of LPH mRNA correlates with the enzymatic activity (r = 0.64). Culture of fetal jejunal explants for 5 days induces by itself a 2‐fold increase in LPH mRNA, without any significant change in lactase enzymatic activity. This increase may reflect the loss of a negative transcriptional regulation operative in vivo, and suggests all additional post‐transcriptional regulatory component. The addition of hydrocortisone (50 ng/ml) during culture induces a doubling of lactase activity without variation in LHP mRNA, indicating a post‐transcriptional modulation by hydrocortisone. The intestinal lysosomal acid β‐galactosidase activity was shown to be unaffected by hydrocortisone treatment. This observation clearly illustrates that the two intestinal β‐galactosidases are regulated differently. Our results suggests a complex developmental regulation of human intestinal lactase and that the perinatal increase in lactase activity could be modulated at a post‐transcriptional level by hydrocortisone.
Characterization of two cDNAs (ERD11 and ERD13) for dehydration‐inducible genes that encode putative glutathione S‐transferases in Arabidopsis thaliana L
Authors: Kiyosue Tomohiro , Yamaguchi-Shinozaki Kazuko , Shinozaki Kazuo
Published: July 2002

Two cDNA clones, designated ERD11 and ERD13, isolated from a cDNA library from Arabidopsis thaliana L. plants dehydrated for 1 h were sequenced and characterized. These clones encoded polypeptides that were homologous to glutathione S‐transferases of tobacco and maize. Genomic Southern hybridization suggested that there are a few additional genes showing high similarity to the ERD11 gene in the Arabidopsis genome. The expression of the genes for ERD11 and ERD13 was induced by dehydration, but was not affected by the application of four plant growth regulators, 2,4‐dichlorophenoxyacetic acid 6‐benzylaminopurine, abscisic acid, or gibberellic acid.
The rat delta‐1 and delta‐2 subunits extend the excitatory amino acid receptor family
Authors: Lomeli H. , Sprengel R. , Laurie D.J. , Köhr G. , Herb A. , Seeburg P.H. , Wisden W.
Published: July 2002

We have characterized a second member (delta‐2) of a new class of subunits for the ligand‐gated excitatory amino acid receptor superfamily. The sequence of delta‐2 exhibits an average identity of 25% and 18.5% to the non‐NMDA and NMDA receptor subunits, respectively. The rat delta‐2 gene is expressed predominantly in Purkinje cells of the cerebellum whereas only low levels of delta‐1 transcripts are found in the adult brain. However, delta‐1 gene expression undergoes a pronounced developmental peak, with particularly high mRNA levels in the caudate putamen of late embryonic/early postnatal stages.
Regulation by protein kinase‐C of putative P‐type Ca channels expressed in Xenopus oocytes from cerebellar mRNA
Authors: Fournier Franck , Charnet Pierre , Bourinet Emmanuel , Vilbert Carole , Matifat Fabrice , Charpentier Gilles , Navarre Pierre , Brûlé Gérard , Marlot Daniel
Published: July 2002

Xenopus oocytes injected with rat cerebellar mRNA expressed functional voltage‐dependent Ca channels detected as an inward Ba current (I
Ba). The pharmacological resistance to dihydropyridines and ω‐conotoxin together with the blockade obtained with Agelenopsis aperta venom suggest that these channels could be somehow assimilated to P‐type Ca channels. The precise nature of the transplanted Ca channels was assessed by hybrid‐arrest experiments using a specific oligonucleotide antisense‐derivated from the recently cloned α1‐subunit of P channels (BI‐1 clone). In addition, we demonstrate that exogenous Ca channel activity was enhanced by two different PKC activators (a phorbol ester and a structural analog to diacylglycerol). The general electrophysiological and pharmacological properties of the stimulated Ca channels remain unchanged. This potentiation induced by PKC activators is antagonized by a PKC inhibitor (staurosporine) and by a monoclonal antibody directed against PKC. It is concluded that P‐type Ca channels are potentially regulated by PKC phosphorylation and the functional relevance of this intracellular pathway is discussed.
The thermodynamics of the unfolding of an isolated protein subdomain The 255–316 C‐terminal fragment of thermolysin
Authors: Conejero-Lara Francisco , De Filippis Vincenzo , Fontana Angelo , Mateo Pedro L.
Published: July 2002

Differential scanning calorimetry has been used to study the thermal unfolding of the 255–316 C‐terminal fragment of thermolysin. The concentration effect on the calorimetric transitions of the fragment in 0.1 M NaCl and 20 mM phosphate buffer, pH 7.5, shows that it behaves as a highly stable dimer in solution, whithin the concentration range 0.19–4.55 mg/ml, undergoing a reversible two‐state thermal unfolding process. The thermodynamic parameters of unfolding (ΔG = 60 ± 6 kJ/(mol of dimer) at 20°C) are similar to those normally observed for small, compact, globular proteins. This and previous studies [1989, Eur. J. Biochem. 180, 513–518] show that the 255–316 fragment folds into a stable, native‐like globular structure.
Bovine seminal plasma ASFP: Localization of disulfide bridges and detection of three different isoelectric forms
Authors: Einspanier Ralf , Krause Ingolf , Calvete Juan J. , Töfper-Petersen Edda , Klostermeyer Henning , Karg Heinrich
Published: July 2002

Acidic seminal fluid protein (aSFP) is a major 13 kDa protein isolated from bull seminal plasma and characterized as a new growth factor which stimulates in vitro cell division and progesterone secretion by ovarian cells. Here, we establish that the four cysteines of oxidized aSFP form two disulfide bridges between nearest‐neighbour residues. This pattern is conserved in boar spermadhesins, with which aSFP shares up to 50% amino acid sequence identity, and other proteins of the recently identified CUB domain family. Using isoelectric focusing in combination with sulfhydryl group‐specific blotting, the three forms of aSFP were identified as completely oxidized (pI 4.7), partly reduced (pI 4.8) and fully reduced at pI 5.1. These results indicate that native aSFP possesses two pairs of cysteine residues of different reactivity. The observation that aSFP can protect sperm from oxidative damage might be explained by its reduction/oxidation behaviour.
Long circulating, cationic liposomes containing amino‐PEG‐phosphatidylethanolamine
Authors: Zalipsky Samuel , Brandeis Ester , Newman Mary S. , Woodle Martin C.
Published: July 2002

Ligand attachment to polyethylene glycol (PEG) grafted, long circulating liposomes at the polymer terminus is of interest for targeting but the effect of positively charged groups is unknown. Amino‐polyethylene glycol‐phosphatidylethanolamine (AminoPEG‐PE), prepared in four steps from α‐amino‐ω‐hydroxy‐PEG, was tested for influence on liposome interactions in vivo: blood circulation and biodistribution. Despite surface amines on each liposome conferring cationic behavior, in vivo properties are comparable to those obtained with methoxy‐PEG‐PE. The consequences are profound for targeting and possibly systemic delivery of cationic lipidic‐polynucleotide complexes.
Purification and characterization of a novel superantigen produced by a clinical isolate of Yersinia pseudotuberculosis
Authors: Yoshino Ken-ichi , Abe Jun , Murata Hiroshi , Takao Toshifumi , Kohsaka Takao , Shimonishi Yasutsugu , Takeda Tae
Published: July 2002

A superantigen designated as Yersinia pseudotuberculosis‐derived mitogen (YPM) was purified in an equal manner from both the culture supernatant and cell lysate of a clinical isolate (KUR‐1) of Y. pseudotuberculosis serotype 4b. A significant proliferative response of human peripheral blood mononuclear cells to purified YPM was detectable even at a concentration of 1 pg/ml. The N‐terminal sequence of YPM which included 23 amino acid residues was determined, by automated Edman degradation, as Thr‐Asp‐Tyr‐Asp‐Asn‐Thr‐Leu‐Asn‐Ser‐Ile‐Pro‐Ser‐Leu‐Arg‐Ile‐Pro‐Asn‐Il e‐Ala‐Thr‐Tyr‐Thr‐Gly‐. This sequence differed from not only all the, hitherto, reported superantigens but also known proteins. While molecular weights of known bacterial superantigens are more than 22,000, electrospray ionization mass spectrometry showed that the molecular weight of YPM was 14524.4. These results indicate that YPM comprises a novel superantigen with substantial structural differences from other bacterial superantigens produced by Gram‐positive cocci.
The CuAsite of the caa3‐type oxidase of Bacillus subtilis is a mixed‐valence binuclear copper centre
Authors: von Wachenfeldt Claes , de Vries Simon , van der Oost John
Published: July 2002

A copper‐containing domain of the caa
3‐type oxidase from Bacillus subtilis has been expressed as a water‐soluble protein in the cytoplasm of Escherichia coli. Electron paramagnetic resonance (EPR) spectra of this purple domain show well‐resolved lines in the g
z resonance, both at X‐band and S‐band frequencies. Interpretation of EPR spectra and analytical data indicate a binuclear copper site consisting of one Cu2+ and one Cu1+. This copper site closely resembles CuA in subunit II of cytochrome c oxidase and is shown here to be a mixed‐valence [Cu2+‐Cu1+) binuclear centre.
Peroxynitrite releases copper from caeruloplasmin: implications for atherosclerosis
Authors: Swain Jason A. , Darley-Usmar Victor , Gutteridge John M.C.
Published: July 2002

Peroxynitrite may be formed in the vasculature by the reaction of puperoxide with nitric oxide. When the blue copper‐containing protein, caeruloplasmin, is incubated with peroxynitrite, copper is released, and ferroxidase activity and the blue colouration are lost. When plasma from normal subjects is incubated with peroxynitrite, the oxidant reacts with numerous plasma constituents but is still able to release copper from caeruloplasmin. As the ferroxidase activity of caeruloplasmin is lost in plasma in the presence of peroxynitrite, a second ferroxidase activity associated with peroxidised lipids, and not inhibited by azide, is formed.
Enzyme from the medicinal leech (Hirudo medicinalis) that specifically splits endo‐ϵ(‐γ‐Glu)‐Lys isopeptide bonds: cDNA cloning and protein primary structure
Authors: Fradkov Arkady , Berezhnoy Sergey , Barsova Ekaterina , Zavalova Lyudmila , Lukyanov Sergey , Baskova Isolda , Sverdlov Eugene D.
Published: July 2002

Earlier we detected a novel enzymatic activity in salivary gland secretion of the medicinal leech, splitting isopeptide bonds between the glutamine γ‐carboxamide and lysine ϵ‐amino group. This activity is due to destabilase. We described its partial amino acid sequence and sequences of two closely related cDNAs, but none of them perfectly matched the protein isolated. Here we report the isolation and sequence peculiarities of the third cDNA of the family as well as the complete sequence of the destabilase protein. The inferred mature protein product of this cDNA matches the independently determined destabilase protein sequence. It contains 115 amino acid residues including 14 highly conserved Cys residues and is formed from a precursor containing specific leader peptide.
Statistical power in operations management research
Authors: Verma Rohit , Goodale John C.
Published: July 2002

This paper discusses the need and importance of statistical power analysis in field‐based empirical research in Production and Operations Management (POM) and related disciplines. The concept of statistical power analysis is explained in detail and its relevance in designing and conducting empirical experiments is discussed. Statistical power reflects the degree to which differences in sample data in a statistical test can be detected. A high power is required to reduce the probability of failing to detect an effect when it is present. This paper also examines the relationship between statistical power, significance level, sample size and effect size. A probability tree analysis further explains the importance of statistical power by showing the relationship between Type II errors and the probability of making wrong decisions in statistical analysis. A power analysis of 28 articles (524 statistical tests) in the Journal of Operations Management and in Decision Sciences shows that 60% of empirical studies do not have high power levels. This means that several of these tests will have a low degree of repeatability. This and other similar issues involving statistical power will become increasingly important as empirical studies in POM study relatively smaller effects.