bluuebunny/crossref_metadata_2025 huggingface dataset:
Engineering and geological features of the soil section of the main water pipeline route in Central Yakutia
Authors: Sal'va Andrei Mikhailovich
Published: April 2020

The object of this study is the permafrost soils of the section of the route of the main water pipeline «Tabaga-Byuteidyakh» in rural areas of Central Yakutia, spanning approximately 28 kilometers. Permafrost soils are widely developed in the Southeastern part of Central Yakutia, and are located on the Leno-Amginsky interfluve. They lie in the upper part of the geological section and are underlying on the lane of the main water pipeline route, as well as fall into the foundations of engineering structures of industrial facilities and hydraulic engineering construction. The study is based on a complex of field engineering and geological surveys, as well as laboratory studies of the physical and mechanical properties of rocks. The article discusses the features of the composition, structure and properties of rocks in this area. It is noted that among the considered soils, there is a variety of granulometric types, cryogenic texture from massive to layered, with clay soils usually of higher ice content than sandy ones. Technogenic loads are also mentioned in the form of objects of the main water supply system, consisting of pumping stations, pipelines, reservoirs and channels, which lead to significant changes in the geocryological environment and surrounding landscapes.
Stable isotopes of carbon in modern plants of tracts of the key terrain of Marre-Sale (Western Yamal)
Authors: Kuznetsova Anna Olegovna , Ivanova Anna Andreevna , Slagoda Elena Adol'fovna , Tikhonravova Yana Vital'evna
Published: April 2020

Studies of the dependence of the accumulation of stable carbon isotopes on the growing conditions in modern plant species of tundra are very relevant. It was discovered that the same plant species accumulate the stable 13C isotope in different ways. The goal of this work is to reveal the relationship between the content of the stable 13C isotope in modern plants and the natural conditions of typical terrains of the tundra of the Western Yamal. At the geocryological station Marre-Sale, modern plant species were selected that grow under conditions of different moisture and remoteness from the sea. Modern plant species were determined using determinant atlases, carbon isotope contents using a DELTA V Advantage isotope mass spectrometer in the laboratory of LBIT (IMCES SB RAS). In different tracts, for identical plants the distribution of carbon isotopes depending on moisture was confirmed: the heavier 13С accumulates in dry ones, and lighter in humid and moist ones. It was determined that near the sea, the same plants of dry and wet tracts accumulate heavier 13С due to the influence of marine aerosols.
Difficulties in the isotopic indication of the genesis Yamal's massive ice. Part 2. Kharasavey
Authors: Chizhova Julia Nikolaevna , Vasil'chuk Yurij Kirillovich
Published: April 2020

Massive ice, located at different depths in the permafrost of the Kharasavey gas condensate field, many massive ice formations occurred in different forms such as layers, lenses and laccoliths. Massive ice near the Kharasavey village was repeatedly studied and tested in detail, and ice formations were found both within the first sea terrace and within the third sea terrace. The isotopic and chemical composition of massive ice can be explained by different ways. We believe that it indicates intrasedimental formation of massive ice. This study is based on data on the values of δ18О, δ2H and dexc in massive ice, as well as the chemical composition of the ice to establish possible conditions for the massive ice formation. Geochemical studies of sediments within the coastal areas of the Kharasavey showed that variability in the distribution of salts in sediments reaches a maximum here, especially in the transition zone from sea to land. The isotopic and chemical characteristics of the ice indicate that ice had been formed in an open system (i.e., with free flow of water from the reservoir). At the same time, the water in the reservoir was changed; at the first stages, it was most likely a mixture of sea and fresh water, which was subsequently desalinated more and more.
Creating an electronic textbook in EXCEL using the VBA programming language
Authors: Romanchenko Tatyana Nikolaevna
Published: April 2020

The author gives the presentation of the content and structural components of an electronic textbook using Excel and the programming language Visual Basic for Applications using an example of a textbook fragment. The methods of presenting theoretical material, modeling with a visual demonstration, automatic verification of the results of assignments, automatic visual display of the results of mastering the topic using the built-in Excel functions are described in detail. The codes of programs for automatic presentation of individual tasks, automatic data modeling based on built-in Excel functions and building a scatter chart with smooth curves are given. The research methodology is based on testing the capabilities of MS Excel and the built-in programming language Visual Basic for Applications to solve the tasks. In contrast to the tools available today for creating electronic educational resources, such a textbook is easily modifiable, allows you to change the scripts of the educational process, in addition to test tasks, present students with sets of individual tasks, and adapt tasks to the level of students. In addition, it is an effective didactic tool for simultaneously mastering the Excel spreadsheet processor and discipline that requires modeling or performing complex calculations and routine operations.
Genesis and functions of the motifs of initiation in the Crimean Tatar epic tale
Authors: Seferova Esma Enverovna
Published: April 2020

The subject of this research became the examination of genesis and functions of the motif of initiation within the Crimean Tatar magic tale. The ritual of initiation is an important social institute of pre-class society, which serves as the foundation of folk tradition for the peoples of the world. The archaic forms of this motif are associated with the mythological rituals-initiations. The motif of initiation in a magic tale often become the core of the plot and compositional structure. Special attention deserves the concept of the scholar Kerim Camanaqlı, who underlined the role of mythological images in development of the storyline of Crimean Tatar tales, as well as primeval roots of these images. The methodology and theoretical framework of the study is the compilations of Crimean Tatar tales of the early XX century “Tales and Legends of Crimean Tatars” (1936) and “Tales for Children” (1939), dictionaries, scientific writings of the prominent folklorists and literary historians. The novelty consists in revelation of the fact that Crimean Tatar magic tales, containing various compositional elements in their structure, include the pair “dying – rebirth”. The indicated elements of the composition of a magic tale emerged and formed under the influence of mythological rituals. By the specific attributes, the ritual of initiation in the tales can carry different functions and forms. Poetic transformation of the motif of initiation defines the peculiarities of magic tales. The main conclusion consists in the ethnographic ritualistic archaic, which vividly manifests in the Crimean Tatar magic tale, inherent with the formation of fundamentals of the tale, its initial structure, situations, images, system of poetic language and artistic means. Mythical character somewhat alienates from previous mythologicity and comes closer to a fairy image, being included into a fairy civilization.
Functional grammar and teaching of Russian grammar in China
Authors: WANG ZHIQIANG , ZHAO XINYI
Published: April 2020

The subject of this research is the implementation of the theory of functional grammar in teaching of Russian language grammar to Chinese audience. The object of this research is the functional grammar of A. V. Bondarenko and communicative function of G. A. Zolotova. Studying the theory of functional grammar of the Russian language and its application among the Chinese audience allows deepening the understanding of grammar rules and increase the efficiency of teaching Russian grammar to Chinese students. The article analyzes the current situation regarding the teaching of Russian grammar in China. Special attention is given to such question as the method of employment of the theory of functional grammar at the grammar lessons among Chinese audience. The novelty consist in the attempt to implement the theory of functional grammar in teaching of Russian language grammar to Chinese students. The article determines the flaws of the traditional structural grammar and merits of the functional grammar. It is revealed that the application of functional grammar would help the students to improve the command of grammar rules and speech communication.
Letter in a system of hypergenre “personal file”
Authors: Kornienko Kristina Borisovna
Published: April 2020

The subject of this research is the structural components of hypergenre “personal file”. The object of this research is business letters. Analysis is conducted on the basis of archived personal filed of the students of Arkhangelsk State Medical Institute for the period from 1930’s – 1940s. The goal consist in introducing new texts of business correspondence into the scientific discourse. In the course of this study, the author solves the following tasks: extraction of documents from the personal files that genre-wise belong to correspondence; determination of genre distinctness of the extracted documents; description of stylistic and functional characteristic of letters and notes. The discursive-communicative analysis of the texts of business correspondence proves that the addresser and the addressee are based on the general social presuppositions. The genre analysis allowed studying the specificity of the genre of business letter and note. For examination of linguistic units of business texts, the author uses the methods of semantic, stylistic and discursive analysis. It is claimed that the hypergenre “personal file” includes a special genre group – business letter or note that manifests as the structural elements of personal file. The author highlights the documents comprising the group of letters/notes based on the formal criterion, i.e. presence of addressee; from the perspective of functional criterion, namely the purpose of document and intention of the addresser; according to discursive-compositional markers, such as form of address or etiquette formula. The article demonstrates the specificity of indicated genres in the particular historical period within the framework of a hypergenre, which is an important addition to the overall picture of the development of business writing, as well as a source for determining normative and stylistic peculiarities of the document on the various language levels.
“Candidatus Trichorickettsia mobilis”, a Rickettsiales bacterium, can be transiently transferred from the unicellular eukaryote Paramecium to the planarian Dugesia japonica
Authors: Modeo Letizia , Salvetti Alessandra , Rossi Leonardo , Castelli Michele , Szokoli Franziska , Krenek Sascha , Serra Valentina , Sabaneyeva Elena , Di Giuseppe Graziano , Fokin Sergei I. , Verni Franco , Petroni Giulio
Published: April 2020

Most of the microorganisms responsible for vector-borne diseases (VBD) have hematophagous arthropods as vector/reservoir. Recently, many new species of microorganisms phylogenetically related to agents of VBD were found in a variety of aquatic eukaryotic hosts; in particular, numerous new bacterial species related to the genus Rickettsia (Alphaproteobacteria, Rickettsiales) were discovered in protist ciliates and other unicellular eukaryotes. Although their pathogenicity for humans and terrestrial animals is not known, several indirect indications exist that these bacteria might act as etiological agents of possible VBD of aquatic organisms, with protists as vectors. In the present study, a novel strain of the Rickettsia-Like Organism (RLO) endosymbiont “Candidatus (Ca.) Trichorickettsia mobilis” was identified in the macronucleus of the ciliate Paramecium multimicronucleatum. We performed transfection experiments of this RLO to planarians (Dugesia japonica) per os. Indeed, the latter is a widely used model system for studying bacteria pathogenic to humans and other Metazoa. In transfection experiments, homogenized paramecia were added to food of antibiotic-treated planarians. Treated and non-treated (i.e. control) planarians were investigated at day 1, 3, and 7 after feeding for endosymbiont presence by means of PCR and ultrastructural analyses. Obtained results were fully concordant and suggest that this RLO endosymbiont can be transiently transferred from ciliates to metazoans, being detected up to day 7 in treated planarians’ enterocytes. Our findings might offer insights into the potential role of ciliates or other protists as putative vectors for diseases caused by Rickettsiales or other RLOs and occurring in fish farms or in the wild.
Proteomic analysis and interactions network in leaves of mycorrhizal and nonmycorrhizal sorghum plants under water deficit
Authors: Olalde-Portugal Víctor , Cabrera-Ponce José Luis , Gastelum-Arellanez Argel , Guerrero-Rangel Armando , Winkler Robert , Valdés-Rodríguez Silvia
Published: April 2020

For understanding the water deficit stress mechanism in sorghum, we conducted a physiological and proteomic analysis in the leaves of Sorghum bicolor L. Moench (a drought tolerant crop model) of non-colonized and colonized plants with a consortium of arbuscular mycorrhizal fungi. Physiological results indicate that mycorrhizal fungi association enhances growth and photosynthesis in plants, under normal and water deficit conditions. 2D-electrophoresis profiles revealed 51 differentially accumulated proteins in response to water deficit, of which HPLC/MS successfully identified 49. Bioinformatics analysis of protein–protein interactions revealed the participation of different metabolic pathways in nonmycorrhizal compared to mycorrhizal sorghum plants under water deficit. In noninoculated plants, the altered proteins are related to protein synthesis and folding (50S ribosomal protein L1, 30S ribosomal protein S10, Nascent polypeptide-associated complex subunit alpha), coupled with multiple signal transduction pathways, guanine nucleotide-binding beta subunit (Rack1) and peptidyl-prolyl-cis-trans isomerase (ROC4). In contrast, in mycorrhizal plants, proteins related to energy metabolism (ATP synthase-24kDa, ATP synthase β), carbon metabolism (malate dehydrogenase, triosephosphate isomerase, sucrose-phosphatase), oxidative phosphorylation (mitochondrial-processing peptidase) and sulfur metabolism (thiosulfate/3-mercaptopyruvate sulfurtransferase) were found. Our results provide a set of proteins of different metabolic pathways involved in water deficit produced by sorghum plants alone or associated with a consortium of arbuscular mycorrhizal fungi isolated from the tropical rain forest Los Tuxtlas Veracruz, México.
Energy dependent interaction of oligomycin and dicyclo hexylcarbodiimide with the mitochondrial membrane
Authors: Azzi Angelo , Santato Mario
Published: July 2002

The characteristics of the oligomycin and DCCD inhibitions of energy linked reactions supported by ATP, indicate that the inhibition is increased by the presence of ATP. It is proposed that such an effect can be attributed to an energy linked change in the structure of some component of the ATPase, facilitating oligomycin interaction with the membrane.
Structure primaire du caseinomacropeptide des caseines κ porcine et humaine
Authors: Chobert Jean-Marc , Mercier Jean-Claude , Bahy Chantal , Hazé Gérard
Published: July 2002

The amino acid sequence of porcine and human caseinomacropeptides (CMP), the C‐terminal glycopeptide released from κ‐casein by chymosin at the initial step of milk coagulation, have been investigated. The complete amino acid sequence of porcine CMP and that of the first 59 amino acid residues of human CMP have been determined. Porcine and human CMPs contain 71 and likely 65 amino acid residues respectively. The extra hexapeptide 38–43 found in porcine CMP arises obviously from the duplication of the DNA fragment coding for the 6 preceding amino acids.
Bromobimane crosslinking studies in oligomycin‐sensitive ATPase from beef heart mitochondria
Authors: Zimmer Guido , Mainka Luise , Heil Berthold M.
Published: July 2002

Using a bromobimane fluorescent label the M
r 31 000 protein band oligomycin‐sensitive (OS)‐ATPase from beef heart mitochondria is shown to become much intensified by 2‐mercaptopropionylglycine. In the presence of 3.5 nmol/mg protein of the thiol reagent ATP—Pi exchange activity is increased by 90%. With the fluorescent crosslinking reagent dibromobimane (DB) we show that a new fluorescent peak appears between M
r 50 000 and 60 000. ATP—Pi exchange is very much decreased by DB. The results suggest that for regulation of ATP‐synthetase activity sulfhydryl groups in the region of the M
r 31 000 protein(s) play an important role.
Reversible effects of cross‐linking on the regulatory cooperativity of Acinetobacter citrate synthase
Authors: Mitchell Colin G. , Weitzman P.D.J.
Published: July 2002

Citrate synthase was purified from Acinetobacter calcoaceticus and treated with the cleavable cross‐linking reagent dithiobis(succinimidyl propionate). Cross‐linking of the enzyme resulted in the abolition of the sigmoidal responses to inhibition by NADH and re‐activation by AMP displayed by the native enzyme. Inhibition and re‐activation were still observed but without any cooperativity. Cleavage of the disulphide bonds in the cross‐links by treatment with dithiothreitol restored the sigmoidal characteristics of both inhibition and re‐activation.
Proton currents and protein motion in membranes
Authors: Marvin D.A.
Published: July 2002

A mechanism is proposed whereby a proton gradient along a membrane‐spanning α‐helix is coupled to small changes in the torsional angles around the α‐helix peptide bonds. Small concerted changes in the torsional angles are coupled to a change in the unit twist of the α‐helix; a change in the unit twist is coupled to a change in the crossing angle between neighbouring α‐helices; and a change in the crossing angle is coupled to a change in the size and shape of an assembly of α‐helices. Following this logical linkage in one direction shows how a proton gradient could induce a pumping motion in an assembly of α‐helices; following it in the other direction shows how motion in an assembly of α‐helices could pump protons.
DNA sequence of the promoter region of the ompC gene and the amino acid sequence of the signal peptide of pro‐OmpC protein in Escherichia coli
Authors: Mizuno Takeshi , Chou Mei-Yin , Inouye Masayori
Published: July 2002

The osmoregulated ompC gene of Escherichia coli was cloned and the DNA sequence of a fragment encompassing the promoter region and a portion of the coding region was determined. There were no obvious homologies in the DNA sequence of the promoter regions of the ompC and ompF genes, in contrast to those of the coding regions of the two genes, both of which code for the matrix porins (major outer membrane proteins) and form passive diffusion pores. The amino acid sequence of the signal peptide of pro‐OmpC protein was also deduced from the DNA sequence
Inhibition by aphidicolin and dideoxythymidine triphosphate of a multienzyme complex of DNA synthesis from human cells
Authors: Wickremasinghe R.Gitendra , Hoffbrand A.Victor
Published: July 2002

A multienzyme complex consisting of DNA polymerase and several DNA precursor‐synthesizing enzymes was solubilized by gentle lysis of cultured human cells. This complex channelled the distal precursor [3H]dTMP into DNA. The patterns of inhibition of the complex by aphidicolin and dideoxythymidine triphopshate (ddTTP) suggested that the complex contained the replicative DNA polymerase, polymerase α. Inhibition by ddTTP was competitive with dTTP. This was exploited to estimate the effective concentration of [3H]dTTP at the site of DNA synthesis during channelling of [3H]dTMP into DNA. The estimated concentration (about 50 μM) was so high as to suggest that the solubilized complex was able to functionally compartmentalize DNA precursors.
Synthesis in vitro of rat brown adipose tissue 32000 Mr protein
Authors: Freeman Karl B. , Chien Sing-Ming , Litchfield David , Patel Hasmukh V.
Published: July 2002

The synthesis of the mitochondrial inner membrane 32 000 M
r protein from rat brown adipose tissue was examined. Polysomes from the tissue were translated in a reticulocyte lysate protein‐synthesizing system and newly‐synthesized protein isolated with a monospecific antibody against the 32 000 M
r protein. The newly‐synthesized protein had the same relative molecular mass as the mature protein. It was taken up by mitochondria isolated from Chinese hamster ovary cells into a form resistant to trypsin.
Karyoduction in Saccharomyces cerevisiae
Authors: Maemura Tohru , Yamashita Ichiro , Fukui Sakuzo
Published: July 2002

We introduced the nuclei isolated from the respiration‐sufficient killer strain of the yeast Saccharomyces cerevisiae into the yeast protoplasts prepared from the respiration‐deficient non‐killer strain with the aid of polyethylene glycol. The resulting karyoductants were respiration‐deficient non‐killers. Nuclear staining with ethidium bromide or DAPI and tetrad analysis of the karyoductants presented evidence that the nuclei introduced into protoplasts were fused to the resident nuclei, leading to stable diploids. This technique termed karyoduction will be useful in the study of nucleo—cytoplasmic relationship in yeast and other organisms.
Catalytic properties of the ATPase on submitochondrial particles after exchange of tightly bound nucleotides under different steady state conditions
Authors: Myers Jill A. , Boyer Paul D.
Published: July 2002

Energized submitochondrial particles were subjected to high or low [3H]ATP/[3H]ADP ratios, maintained during steady state by a pyruvate kinase or hexokinase regenerating system, respectively. Under both steady state conditions, about 1.4 mol [3H]nucleotide/mol ATPase was retained but considerably more [3H]ATP was retained with the high [3H]ATP/[3H]ADP ratio. The ATPase activity and the oxygen exchange of these differentially labeled SMP were the same, suggesting a lack of control function of non‐catalytic tightly bound nucleotides.
Studies on the mechanism of inhibition of hepatic cAMP accumulation by vasopressin
Authors: Morgan Noel G. , Shipp Clanton C. , Exton John H.
Published: July 2002

Vasopressin elicited a dose‐dependent inhibition of glucagon‐induced cAMP accumulation in isolated hepatocytes. This response was not diminished by incubation of cells with the calmodulin antagonists trifluoperazine or chlorpromazine and was only slightly reduced in Ca2+‐depleted hepatocytes. Half‐maximal inhibition of cAMP accumulation occurred at 8 × 10‐11 M vasopressin, a dose which does not increase cytosolic Ca2+ in hepatocytes. Direct activation of adenylate cyclase by forskolin was significantly inhibited by vasopressin in Ca2+‐depleted cells. It is concluded that inhibition of hormone‐induced cAMP accumulation by vasopressin in liver is not dependent on cellular Ca2+ mobilisation but may involve direct inhibition of adenylate cyclase.
Calcium uptake during the cell cycle of Saccharomyces cerevisiae
Authors: Saavedra-Molina Alfredo , Villalobos Rafael , Borbolla Miguel
Published: July 2002

Synchronous culture of the budding yeast Saccharomyces cerevisiae was obtained by sucrose density gradient selection with 90–100% of yeast synchronized by using the cells in the bottom. In these adult cells bud emergence is coincident with an increase in calcium uptake at 100 min of the culture, followed by a return to basal values which are maintained until the end of the first cell cycle of study. The phenothiazine derivatives, trifluoperazine and chlorpromazine inhibit bud emergence and trifluoperazine also increases calcium uptake.
Equilibration of methionine into the Met‐tRNA pool of rabbit reticulocyte lysates
Authors: Gross Martin , Kaplansky Daniel A. , Knish Walter M.
Published: July 2002

We have measured the rate of equilibration of [35S]methionine into the Met‐tRNA pool of rabbit reticulocyte lysate as in [FEBS Lett. (1982) 143, 301–305]. Our results indicate that hemin‐deficiency inhibits the equilibration of methionine into the tRNA pool much less than protein synthesis or the equilibration of alanine into the tRNA pool, whereas cycloheximide inhibits these processes similarly. This finding is consistent with our previous data and supports the hypothesis that with hemin‐deficiency much of the Met‐tRNAf that becomes bound to 40 S subunits subsequently undergoes enzymatic deacylation.
Template‐independent synthesis of guanosine tetra‐ and pentaphosphates on ribosomes
Authors: Belitsina Nadezhda V. , Klyachko Elena V. , Shakulov Rustem S.
Published: July 2002

It is shown that Escherichia coli ribosomes carrying poly(Lys)‐tRNA can form (p)ppGpp in the presence of stringent factor in the absence of the poly(A) template. Template‐independent synthesis of (p)ppGpp is suppressed by tetracycline and partially decreases if deacylated tRNA is omitted.
EPR titration of ovine prostaglandin H synthase with hemin
Authors: Ruf H.H. , Schuhn D. , Nastainczyk W.
Published: July 2002

To characterize further the prosthetic group of PGH synthase (EC 1.14.99.1), titrations of the apoenzyme with hemin were investigated by EPR. The first hemin bound per polypeptide showed an EPR signal at g = 6.7 and 5.3 (rhombicity 9%) and was tentatively assigned to the hemin effective as prosthetic group of PGH synthase. Additional hemin bound showed a less rhombic signal (g = 6.3 and 5.8, rhombicity 3%) presumably due to nonspecific hydrophobic binding sites not effective in catalysis.
Effects of FSH on acidic nuclear protein synthesis in cultured pig Sertoli cells
Authors: Perrard M.H. , Saez J.M. , Dazord A.
Published: July 2002

We studied the effect of FHS on nuclear protein synthesis by a primary culture of immature porcine Sertoli cells. The cells were cultured in a chemically defined medium and treated with FSH (50 ng/ml) for various periods (4, 8, 25 and 64 h). About 65 spots (pHi < 8) were identified by two‐dimensional polyacrylamide gel electrophoresis of the radiolabelled nuclear proteins. After 25 h of FHS treatment, we observed an increase of 5 proteins, a slight decrease of two and a disappearance of one. For the short periods of FSH treatment (4 and 8 h), no effect of FHS on nuclear protein synthesis was observed. After 64 h of FSH treatment, the synthesis of all nuclear proteins appears to be decreased.
The genome of Theileria parva: some structural properties
Authors: Guerin-Marchand Claudine , Lambiotte Maurice
Published: July 2002

The DNA of the protozoan Theileria parva, the causal agent of the bovine East Coast Fever, has been prepared at least 99% pure from the intra‐erythrocytic form of the parasite. Its buoyant density was found to be 1.696 g/cm3 and its calculated G + C content was 36.7%. Fragmentation of T. parva by the restriction enzyme EcoRI provides some evidence of the presence of repetitive DNA sequences.
Cloning and expression in Escherichia coli of the insecticidal δ‐endotoxin gene of Bacillus thuringiensis var. israelensis
Authors: Ward E.S. , Ellar D.J. , Todd J.A.
Published: July 2002

Recombinant plasmids containing the mosquitocidal δ‐endotoxin gene were constructed by inserting HindIII fragments of the Bacillus thuringiensis var. israelensis 72–75 Md plasmid in to the Escherichia coli vector pUC12. Two recombinants producing the 26000 Da δ‐endotoxin (pIP173 and pIP174) were identified by screening clones in an E. coli in vitro transcription‐translation system. Both recombinants were 12.4 kb chimaeric plasmids comprising pUC12 and a common 9.7 kb HindIII fragment of the B. thuringiensis plasmid. The 26000 Da polypeptide synthesis in vivo from pIP174 transformed into E. coli JM101 was lethal to mosquito larvae and cytotoxic to mosquito cells in vitro. The biological authenticity of the cloned product was further confirmed by demonstrating that the cytotoxicity of the polypeptide was neutralised by antiserum to the authentic δ‐endotoxin or by preincubation with excess toxin receptor. Transcription of the recombinant δ‐endotoxin gene in E. coli appears to utilise a Bacillus promotor sequence(s) rather than the pUC12 β‐galactosidase promotor.
Increased insulin sensitivity in soleus muscle from cold‐exposed rats: reversal by an adenosine‐receptor agonist
Authors: Budohoski Leszek , Challiss R.A.John , Lozeman Fred J. , McManus Bronwyn , Newsholme Eric A.
Published: July 2002

The effect of 0.5,2,7 and 14 days cold exposure at 4°C on insulin sensitivity was investigated in the stripped soleus muscle preparation incubated in vitro. Cold‐exposure for 2 or 7 days increased the sensitivity of glycolysis, but did not affect the sensitivity of glycogen synthesis to insulin. Cold‐exposure for 0.5 or 14 days had no effect on the sensitivity of either process to insulin. The increased sensitivity to insulin after exposure of animals to the cold for 2 days was completely reversed by addition of the adenosine receptor agonist, 2‐chloroadenosine, to the incubation medium. This suggests that cold exposure may increase insulin sensitivity in the muscle, either by a decrease in the concentration of adenosine in the muscle, or by a decrease in the number or affinity of the adenosine receptors.
5 S RNA‐like structures in large ribosomal subunit RNAs of fungal mitochondria
Authors: Thurlow David L. , Mason Thomas L. , Zimmermann Robert A.
Published: July 2002

5 S ribosomal RNA
Mitochondrial ribosome
Protein synthesis
Saccharomyces cerevisiae
Aspergillus nidulans
The recognition sequence of site‐specific endonuclease BbvII from Bacillus brevis 80
Authors: Matvienko N.I. , Pachkunov D.M. , Kramarov V.M.
Published: July 2002

Site‐specific endonuclease BbvII from Bacillus brevis 80 recognizes the non‐symmetrical hexanucleotide and cleaves DNA at distances of 2 and 6 nucleotides from the recognition site: 5'‐GAAGAC2N↓ 3'‐CTTCTG6N↑
Evidence for a homogeneous lateral distribution of lipids in a bacterial membrane
Authors: de Bony J. , Martin G. , Welby M. , Tocanne J.F.
Published: July 2002

A new photo cross‐linking method has been developed for the study of the lateral distribution of lipids in natural membranes, which uses anthracene as a photoactivable group. This method, which rests on the potentiality of anthracene to form covalently bound dimers upon irradiation around 340–380 nm has been applied to the membrane lipids (dimannosyl diacylglycerol, phosphatidylglycerol, phosphatidylinositol) of the bacterium Micrococcus luteus. These glyco‐ and phospholipids were anthracene labelled by metabolically incorporating the synthetic 9‐(2‐anthryl)nonanoic acid. The following sequential procedure was used: (i) dimerization of the anthracene‐labelled lipids in the membrane by irradiation of the intact cells at 360 nm; (ii) extraction of the lipids and thin‐layer chromatography in the first dimension to separate the various lipid dimers from the monomers; (iii) partial dedimerization of the lipid dimers by illumination of the chromatogram at around 250–280 nm; (iv) chromatography in the second dimension to separate the native lipid monomers from the corresponding residual lipid dimers. On account of the occurrence of the 3 hetero dimers phosphatidylglycerol‐dimannosyl diacylglycerol, phosphatidylinositol‐dimannosyl diacylglycerol and phosphatidylglycerol‐phosphatidylinositol after irradiating the cells, it is concluded that in this bacterial membrane, dimannosyl diacylglycerol, phosphatidylglycerol and phosphatidylinositol are homogeneously distributed.
Inhibitory effect of calmodulin inhibitors on palytoxin‐induced K+ release from rabbit erythrocytes
Authors: Nagase Hiromi , Ozaki Hiroshi , Urakawa Norimoto
Published: July 2002

Palytoxin (PTX) caused K+ release from rabbit erythrocytes at a concentration as low as 10−11 M. The K+ release due to PTX at a concentration below 10−9 M was dependent on Ca2+ in medium. The effect of Ca2+ was substituted fully by Sr2+ and partially by Ba2+. W‐7 (2 × 10−4 M), a known inhibitor of calmodulin, markedly inhibited the rate of K+ release due to PTX. W‐5 (2 × 10−4 M), an analog of W‐7 with lower affinity to calmodulin than W‐7, showed weaker inhibition. Other calmodulin antagonists, such as prenylamine, chlorpromazine and compound , also inhibited the PTX‐induced K+ release. These results suggest that the K+ release induced by PTX involves the process(es) mediated by intracellular Ca2+ and calmodulin.
Hormonal effects on the phosphorylation of glycogen synthase in rat hepatocytes
Authors: Ariño Joaquín , Mor Angels , Fátima Bosch , Baanante Isabel V. , Guinovart Joan J.
Published: July 2002

The effects of epinephrine and vasopressin on the phosphorylation state of glycogen synthase were studied using rat hepatocytes incubated with 32P. After the incubation with hormones, 32P‐labeled glycogen synthase was isolated using antibodies against rat liver enzyme. The immunoprecipitate showed a single radioactive band (M
app 88 kDa) when subjected to SDS‐gel electrophoresis. Both epinephrine and vasopressin inactivated the enzyme and increased the 32P content of glycogen synthase. Cleavage of the immunoprecipitate with CNBr yielded two major 32P‐labeled fragments of M
app∼27 and 12 kDa. Both hormones increased the 32P content of both fragments. These results prove that epinephrine and vasopressin increase the phosphate content of the enzyme promoting its phosphorylation at multiple sites.
Hormonal activation of single K+ channels via internal messenger in isolated pancreatic acinar cells
Authors: Suzuki Kazuo , Petersen Carl C.H. , Petersen Ole H.
Published: July 2002

The mechanism underlying hormonal activation of potassium channels was investigated in pig pancreatic acinar cells by patch‐clamp single‐channel and whole‐cell current recordings. It was shown directly that a peptide hormone belonging to the cholecystokinin‐gastrin family, CCK5, can activate single voltage‐sensitive potassium channels which can be blocked by tetraethylammonium. The single‐channel currents were recorded from electrically isolated cell‐attached membrane patches to which the hormone had no access and the activation must therefore involve an intracellular messenger. The hormonal response requires external Ca2+ in the isolated membrane‐patch area indicating that calcium gating is not directly linked to hormone‐receptor interaction.
The effect of GTP hydrolysis and transpeptidation on the arrangement of aminoacyl‐tRNA at the A‐site of Escherichia coli 70 S ribosomes
Authors: Vladimirov S.N. , Graifer D.M. , Karpova G.G. , Semenkov Yu.P. , Makhno V.I. , Kirillov S.V.
Published: July 2002

From the affinity labelling of 70 S ribosomes with a photoreactive derivative of Phe‐tRNAphe bearing an arylazido group on guanine residues, it has been found that different sets of ribosomal proteins are labelled in the course of three successive steps of EF‐Tu‐dependent binding of aminoacyl‐tRNA derivative at the A‐site. Proteins S5, S7, S8, S16, S17, L9, L14, L15 and L24 were labelled before GTP hydrolysis; proteins S5, S7, S9, S11, S14, S18, S19, S21, L9, L21 and L29 ‐ after GTP hydrolysis; proteins S2, S5, S7, S21, L11 and L23 ‐ after GTP hydrolysis and transpeptidation.
Rapid down‐regulation of protein kinase C and membrane association in phorbol ester‐treated leukemia cells
Authors: Wickremasinghe R.Gitendra , Piga Andrea , Campana Dario , Yaxley John C. , Hoffbrand A.Victor
Published: July 2002

Peripheral blood lymphocytes from patients with chronic lymphocytic leukemia (CLL) acquire after several days of exposure to 12‐O‐tetradecanoylphobol‐13‐acetate (TPA) several morphological, immunological and histochemical features of hairy cell leukemia. We have investigated the short term effects of TPA treatment on protein kinase C and its subcellular distribution. Within minutes of addition of TPA to CLL cells 20% of the cytosolic protein kinase C had associated with the particulate fraction. The remaining 80% of protein kinase C activity was down‐regulated. The association with the membrane dramatically increased the resistance of the enzyme to inhibition by the non‐ionic detergent, Triton X‐100. These results suggest that activation of protein kinase C causes multiple biological changes in CLL cells.
Mutant species of EF‐Tu, altered at position 375, exhibit a reduced affinity for aminoacylated transfer‐RNAs
Authors: Sam Tom , Pingoud Alfred , Bosch Leendert
Published: July 2002

The interaction between EF‐Tu GTP and aminoacyl‐tRNA is shown to be influenced by mutations at site 375 of this three‐domain protein. Site 375 is located in domain II near the interface with domain I [(1984) EMBO J. 3, 113‐120]. Replacement of the alanine at this site by a threonine or valine residue results in lower binding constants with Phe‐tRNA and Tyr‐tRNA, as was evaluated by the hydrolysis protection technique. The data are discussed in the light of what is known about the three‐dimensional structure of the protein and its interaction sites with aminoacyl‐tRNA.
Sodium‐induced conformation changes in membrane transport proteins
Authors: Charalambous Bambos M. , Wheeler Kenneth P.
Published: July 2002

In the presence of KC1, tryptic digestion of vesicles derived from pigeon erythrocyte membranes inactivates sodium‐dependent uptake of alanine by the vesicles, whereas digestion in the presence of NaCl does not. Extensive degradation of vesicle proteins occurs under both conditions. Similarly, the extent of inhibition by N‐ethylmaleimide of the sodium‐dependent influxes of both glycine and alanine into human erythrocytes is greater when the cells are exposed to the thiol reagent in the presence of KC1 than when NaCl is used. These observations are interpreted as providing evidence for sodium‐induced conformation changes in these transport proteins.
On the prosthetic group(s) of component II from nitrogenase
Authors: Hagen W.R. , Dunham W.R. , Braaksma A. , Haaker H.
Published: July 2002

The EPR spectrum of the reduced Fe‐protein from nitrogenase has been reinvestigated. The dependences on temperature, microwave power, and microwave frequency all suggest that the observed signal represents a magnetically isolated [4Fe‐4S]1+(2+;1+) cluster. Also, the signal can be simulated assuming a simple, gstrained S = system. However, the integrated intensity amounts to no more than 0.2 spins per protein molecule. It is, therefore, impossible that Fe‐protein preparations contain a single type of [4Fe‐4S] cluster.
Murine interleukin 1 stimulates α2‐macroglobulin synthesis in rat hepatocyte primary cultures
Authors: Bauer Joachim , Weber Wolfgang , Tran-Thi Thuy-Anh , NorthofF Gerd-Hinrich , Decker Karl , Gerok Wolfgang , Heinrich Peter C.
Published: July 2002

Interleukin 1
α2‐Macroglobulin
Rat hepatocyte
Purification of two species of exudate cysteine‐proteinase inhibitors that are acute‐phase reactants in the carrageenin‐induced inflammation in rats
Authors: Nakagawa Hideo , Hirata Kazunari
Published: July 2002

Two species of cysteine‐proteinase inhibitors (CPIs) have been purified to homogeneity from exudate in the carrageenin‐induced inflammation in rats. The exudate CPIs were separated into two forms (named CPI‐1 and ‐2) in affinity chromatography on S‐carboxymethyl‐papain‐Sepharose, the final stage of purification. CPI‐1 and ‐2 gave different mobilities in polyacrylamide gel electrophoresis (PAGE), probably because of different isoelectric points (pI 4.47 for CPI‐1 and pI 4.21 for CPI‐2). Both CPI‐1 and ‐2 showed immunological identity in double immunodiffusion and same molecular mass of 68 kDa when analysed by SDS‐PAGE. These results indicate that CPI‐1 and ‐2 are very similar but distinct CPIs. CPI‐1 and ‐2 are acute‐phase reactants and probably represent two species of T‐kininogens having inhibitory activity toward cysteine proteinases.
Phosphorylation of the modulator protein of the ATP,Mg‐dependent protein phosphatase by casein kinase TS
Authors: Agostinis Patrizia , Goris Jozef , Vandenheede Jackie R. , Waelkens Etienne , Pinna Lorenzo A. , Merlevede Wilfried
Published: July 2002

The phosphorylation by casein kinase TS (II) of the modulator protein of the ATP,Mg‐dependent phosphatase increases after preineubation with the PCSH1 phosphatase or with the catalytic subunit of the ATP,Mg‐dependent phosphatase. Dephosphorylation by the two phosphatases combined leads to the incorporation of 2 mol phosphate per mol modulator (at Ser residues). Occupancy of the ATP,Mg‐dependent phosphatase phosphorylation site(s) is a negative determinant in the phosphorylation of the modulator by kinase TS. Among the PCS phosphatases PCSh1 shows the highest activity toward the 32P‐Ser residues labeled by kinase TS in untreated or previously dephosphorylated modulator, while the ATP,Mg‐dependent phosphatase is totally ineffective. Protamine stimulates all phosphatase activities, so that the catalytic subunit of the ATP,Mg‐dependent phosphatase becomes almost as effective as the PCSC phosphatase in dephosphorylating the kinase TS sites.
The activity of the Na+/H+ antiporter in cultured cardiac cells is dependent on the culture conditions used
Authors: Green Richard D. , Frelin Christian , Vigne Paul , Lazdunski Michel
Published: July 2002

When chick cardiac cells are maintained in serum‐free NCTC‐135 medium, the pHi dependence of the Na+/ H+ exchanger is much more acidic (pK = 7.0) than when a serum‐free M199 medium is used (pK > 7.5). Phorbol esters activate the Na+/H+ exchanger and produce a cell alkalinization in cells maintained in NCTC‐135 medium by shifting the pHi dependence of the system to more alkaline values. They have no action on cells maintained in M199 medium. The alkaline pHi dependence of the Na+/H+ exchanger and the loss of responsiveness of phorbol ester action in M199 medium correlate with increased rates of phosphatidylinositol turnover.
The effect of substrates on the inter‐domain interactions of the hinge‐bending enzyme 3‐phosphoglycerate kinase
Authors: Adams Benjamin , Pain Roger H.
Published: July 2002

A hinge‐bending mechanism has been proposed for phosphoglycerate kinase, in which the two domains bend about the connecting “swaist& rsquo; region. In partially denaturing concentrations of guanidinium chloride the substrate 3‐phosphoglycerate stabilises one domain against denaturation and destabilises the other. The reduction of mutual stabilisation of the two domains on binding substrate indicates a freeing of the hinge to allow the protein to take up other states rather than a directive mechanism. The stabilisation of both domains at higher concentrations of ATP at which the enzyme is inhibited supports this mechanism.
A monoclonal antibody specific for snRNPs U1 and U2
Authors: Reuter Rolf , Lehner Christian F. , Nigg Erich A. , Lührmann Reinhard
Published: July 2002

A monoclonal antibody (D‐5) is described which selectively precipitates snRNPs U1 and U2. The antibody was derived from a mouse immunized with extracts from chick embryonic nuclei. By immunoblotting on either total proteins from purified snRNPs U1–U6, U2–U6 or U1 only, we could demonstrate that the monoclonal antibody cross‐reacts with the U1 RNP specific polypeptide A and the U2 RNP specific polypeptide B'', thereby establishing that the two snRNP proteins share at least one epitope. D‐5 precipitates snRNPs U1 and U2 from a variety of species, including man, chicken, mouse, rat kangaroo and Xenopus laevis. It will thus be a useful tool for studying structure function relationships of the two snRNP species in different cell systems.
Laser‐activated carbene labels the same residues in the proteolipid subunit of the ATP synthase in energized and nonenergized chloroplasts and mitochondria
Authors: Weber H. , Junge W. , Hoppe J. , Sebald W.
Published: July 2002

The membrane‐embedded proteolipid of the Fo part of the ATP synthase in mitochondria (su 9) and chloroplasts (CFoIII) was labeled with a carbene generated from the photoactivatable hydrophobic reagent [125I]‐TID by a single UV‐laser pulse. Amino acid sequence analysis of the isolated proteolipid revealed that the carbene modified distinct residues which apparently are accessible from the lipid phase. No differences were detected in the labeling pattern upon energization of the membrane. Labeling of mitochondrial proteolipid in the presence of oligomycin led to a reduced modification of several side chains which map directly a surface involved in oligomycin binding.
Isolation of heparin‐binding growth factors from dogfish (Mustela canis) brain and retina
Authors: Lagente O. , Diry M. , Courtois Y.
Published: July 2002

We have purified two growth factors from dogfish brain and retina by using their binding affinity for heparin‐Sepharose. These growth factors were eluted at 1 and 2 M NaCl similarly to those purified from bovine brain or retina. Their mitogenic activity was assayed in vitro on the same mammalian cells: bovine lens epithelial cells or human fibroblasts. All these data seem to indicate that these growth factors belong to the families of other well defined mammalian growth factors: EDGF I, brain basic FGF, AGF II, on the one hand and EDGF II, brain acidic FGF, AGF I, ECGF, on the other. Thus, these growth factors have been widely distributed during evolution and retain at least a conservative sequence to stimulate cell proliferation, in mammalian cells.
The 66 kDa component of eukaryotic initiation factor 3 interacts with globin mRNA and 18 S rRNA in preinitiation complexes
Authors: Westermann P. , Sohi M.K. , Arnstein H.R.V.
Published: July 2002

The 66 kDa protein present in a complexwith globin mRNA and 18 S rRNA [(1984) Eur. J. Biochem. 143, 27‐33] has been reincorporated into functional eukaryotic initiation factor 3 (eIF‐3) under conditions of protein synthesis. Additionally, two‐dimensional polyacrylamide gel electrophoresis has been used to demonstrate the identity of the 66 kDa protein with the 66 kDa subunit ofeIF‐3.
Nucleotide binding to elongation factor 2 inactivated by diphtheria toxin
Authors: Burns Gary , Abraham Abraham K. , Vedeler Anni
Published: July 2002

Binding of guanosine nucleotides to purified native and ADP‐ribosylated wheat germ EF‐2 was measured. Both forms of EF‐2 bound [3H]GDP to the same extent. [3H]GDP binding to native but not to ADP‐ribosylated EF‐2 was reduced in the presence of GTP and ribosomes. Binding of [γ‐32P]GTP to EF‐2 was significantly reduced upon ADP‐ribosylation. ADP‐ribosylation almost abolished both the stimulatory effect of ribosomes on GTP binding to EF‐2 and the ability of EF‐2 to form a high‐affinity complex with GuoPP(CH2)P and ribosomes. Low‐affinity complex formation between EF‐2 · GDP and ribosomes was not influenced by ADP‐ribosylation. The results indicate that the inhibition of the elongation process caused by the toxin is probably due to the inability of modified EF‐2 to exchange GDP with GTP.
The human insulin‐like growth factor II gene contains two development‐specific promoters
Authors: de Pagter-Holthuizen P. , Jansen M. , van Schaik F.M.A. , van der Kammen R. , Oosterwijk C. , Van den Brande J.L. , Sussenbach J.S.
Published: July 2002

The insulin‐like growth factors (IGF) play an important role in fetal and postnatal development. Recently, the nucleotide sequences of the cDNAs encoding IGF‐I and IGF‐II and part of the human IGF genes were reported. In this communication we describe two distinct IGF‐II cDNAs isolated from a human adult liver and a human hepatoma cDNA library, respectively. Using these two cDNAs, we have established that the human IGF‐II gene contains at least 7 exons. Two different IGF‐II promoters have been identified, 19 kilobases (kb) apart, which are active in a development‐specific manner. The promoter, active in the adult stage, is located only 1.4 kb downstream from the insulin gene.