bluuebunny/crossref_metadata_2025 huggingface dataset:
Estudo Prospectivo ao Uso da Farinha do Mesocarpo do Coco Verde como Matéria-Prima para Elaboração de Alimentos
Authors: Da Silva Luís Paulo Firmino Romão , Moraes Maria Suiane de , Esmero Janaína Almeida Dantas , Silva Semirames Do Nascimento , Tomiyoshi Carlos Minor
Published: October 2020

A utilização da farinha do mesocarpo do coco verde como matéria-prima na elaboração de produtos alimentícios, considerando sua composição nutricional e características sensoriais, se destaca, principalmente, pelo elevado teor de fibras, que podem ser utilizadas como aditivo em produtos de panificação, sem alterar suas propriedades sensoriais. O trabalho foi realizado com o objetivo de levantar dados de depósitos de patentes sobre a utilização da farinha do coco verde por meio do mapeamento nas bases do INPI, Espacenet e USPTO. A pesquisa foi realizada em agosto de 2017, usando-se as palavras-chave “coco, produtos e farinha”, em português e em inglês. Os resultados mostraram que os depósitos de patentes que associam a utilização da farinha do mesocarpo do coco na produção de alimentos são escassos, justificando a necessidade de mais pesquisas nessa área, pois, além de poder agregar valor nutricional e econômico ao produto, isso pode diminuir a geração de resíduos provenientes do uso do coco, incentivando o investimento em ações sustentáveis.
Propriedade Intelectual em Empresas Públicas: uma análise dos depósitos de marcas da Embrapa
Authors: SOARES PRETO GLAUBER ADENIR , Franco Cíntia Brenner  Franco Brenner Acosta , Bruch Kelly Lissandra
Published: October 2020

A proposta deste artigo é mostrar os aspectos relevantes acerca do registro de marcas de uma empresa pública no Brasil, em especial, o panorama da Empresa Brasileira de Pesquisa Agropecuária (Embrapa). O objetivo é analisar os dados que permeiam os pedidos de registro de marca no Brasil, as questões sobre como a empresa trabalha e a gestão desses ativos intangíveis para o desenvolvimento da empresa e do ambiente em que ela está inserida. Para o desenvolvimento da pesquisa, foi realizado um estudo de caso, de natureza exploratória e descritiva, com abordagem mista (quanti-qualitativa), por meio de entrevistas de colaboradores da instituição e por pesquisa nas principais bases de dados referentes à propriedade intelectual, entre os anos de 1970 até 2018, utilizando-se como termo de busca a denominação “Embrapa” na titularidade. Todas as plataformas pesquisadas apresentaram número crescente de pedidos ao longo dos anos. Na base do INPI, foram encontrados 484 pedidos de registro de marcas, sendo em sua maioria marcas nominativas. Pode-se concluir que é perceptível o avanço na proteção de marcas pela Embrapa, esse avanço pode se dar devido aos investimentos e às ações do mercado. Todavia, apesar de a empresa possuir um processo para encaminhamento de registro de marcas relativamente conhecido, esse procedimento não está documentado e verificou-se a necessidade de se realizar a documentação do processo, bem como a orientação e a capacitação das equipes para proteção e execução de forma correta dos procedimentos de registros de marcas junto ao INPI.
An alpha2‐globulin component present in sweat, saliva, tears, human milk, colostrum and cerumen
Authors: Jirka M.
Published: July 2002

By double‐diffusion and immunoelectrophoresis, using rabbit anti‐sweat protein immune serum, an alpha2‐globulin component of glycoprotein character was found in sweat, saliva, tears, human milk, colostrum and cerumen.
Les hémoglobines des amphibiens: Caractérisation préliminaire des chaînes d'une hémoglobine de la grenouille Rana esculenta (L)
Authors: Chauvet Jean Pierre , Acher Roger
Published: July 2002

A frog (Rana esculenta) hemoglobin has been purified and two types of chain have been separated by countercurrent distribution, N‐ and C‐terminal sequences of each chain have been determines. The N‐terminal amino acids are Acetyl‐Ala and Gly. The acetylated chain seems to belong to the α‐chain series and the glycine chain to the β‐chain series.
The progressive effects of fasting on glucose phosphorylation by isolated rat hepatocytes: the involvement of a high K0.5 enzyme
Authors: Singh Jasbir , Nordlie Robert C.
Published: July 2002

The effects of varied durations of food deprivation on the rates and kinetics of glucose phosphorylation by isolated rat hepatocytes have been examined. Glucokinase activity was measured concurrently in extracts from these cells prepared from livers of rats which had fasted for 0, 24, 48 and 72 h. Significant levels of hepatocyte glucose phosphorylation were noted even when glucokinase levels were extrapolated to zero. The K
0.5‐glucose value of 33 mM in cells from fed rats increased to 48 mM after a 72‐h fast. It is concluded that a high K
0.5 glucose‐phosphorylating enzyme or enzymes compensatory to insulin‐dependent glucokinase is/are involved in rat liver glucose phosphorylation.
Identification of N‐ and C‐terminal corticotropin peptides in the Mr 80 000 form of neurophysin
Authors: Garnier D. , Morel A. , Beguin P. , Cohen P.
Published: July 2002

The 125I‐labeled M
r 80 000 form of neurophysin has been purified from bovine neurohypophysi. Tryptic digests of this species were analyzed, prior to or after treatment with carboxypeptidase B, by high‐pressure liquid chromatography followed by isoelectric focusing and the fragments compared with those generated by a similar treatment of reference bovine 1–39 adrenocorticotropin. The ACTH peptides 22–39 and 1–8, as well as the 1–7 derivative of the latter were identified by those two independent criteria. This provides chemical evidence supporting the hypothesis [8] that high M
r neurophysin may contain the sequence of ACTH.
The mandatory role of complement in the endotoxin‐induced synthesis of tissue thromboplastin in blood monocytes
Authors: Østerud Bjarne , Eskeland Trond
Published: July 2002

Fres isolated blood cells recombined with normal heparinized plasma and then incubated with endotoxin, induced a 100‐fold increase in monocyte tissue thromboplastin synthesis. In contrast, recombination of these cells with heat inactivated plasma, cobra venom factor‐treated plasma, Ca2+‐free plasma, or BioRex 70‐treated plasma (plasma free of Clq and D) before incubation with endotoxin, failed to induce monocyte synthesis of tissue thromboplastin. These results strongly support the hypothesis that complement is required for endotoxin stimulation of blood monocyte synthesis of tissue thromboplastin.
Dansylsphingolipids for labelling and probing membranes
Authors: Huang Richard T.C.
Published: July 2002

Three fluorescent sphingolipids have been prepared, which contain dansyl sphingosine in the hydrophobic portion and various head groups (F‐Phosphorylcholine, galactose and lactose) in the hydrophilic end. These dansylphingolipids are readily incorporated into membranes and show affinity toward several membrane proteins. Results of fluorescence measurements indicate, that these lipids may be useful as probes for studying protein—lipid interactions.
Hydrophobic chromatography of fibronectin
Authors: Morgenthaler J.-J.
Published: July 2002

Fibronectin was chromatographed on immobilized alkanes of various chain lengths. No binding of the protein to the hydrophobic matrix was observed with alkanes containing from 3–7 methylene groups; the protein bound, however, to immobilized alkanes with 8 or 10 methylene groups. Fibronectin could be quantitatively eluted from commercial Octylsepharose with a non‐ionic detergent. A gelatin‐based plasma expander did not interfere with the binding. Many proteolytic fibronectin fragments also bound to a hydrophobic matix. The results show that fibronectin posesses at least one hydrophobic binding site.
The turnover of vimentin in Ehrlich ascites tumour cells
Authors: McTavish Catherine F. , W. James Nelson , Peter Traub
Published: July 2002

The turnover of vimentin and vimentin‐derived peptides has been examined in logarithmically growing Ehrlich ascites tumour cells. Cells were pulse‐labelled with [35S]methionine for 30 min and then chased for up to 60 h. It was found that the specific radioactivity of the main isoelectric variant of vimentin decreased to half the original value in 15.3 h which was close to the division time of the cells (16 h). The protein moiety of the phosphorylated variant of vimentin also turned over very slowly, in contrast to the turnover rate of the phosphate group itself which has a half‐life of 1.4 h. The role of the intermediate filament‐specific, Ca2+‐activated proteinase has been considered in relationship to the slow turnover of vimentin.
Effect of transmethylation‐reaction and increased levels of cAMP on superoxide generation of guinea‐pig macrophages induced with wheat germ agglutinin and phorbor myristate
Authors: Yagawa Katsuro , Itoh Tokuo , Tomoda Akio
Published: July 2002

Superoxide (O2
−) generation of guinea‐pig macrophages induced by wheat germ agglutinin (WGA) was suppressed to a great extent by the inhibition of transmethylation with 3′‐deazaadenosine. When macrophages were stimulated with phorbor myristate (PMA) instead of WGA, the suppression of O2
− generation of macrophages was observed to be slight despite the presence of 3′‐deazaadenosine. These results were confirmed under various conditions. Thus the WGA‐stimulated O2
− generation of macrophages is probably associated with transmethylation, but the PMA‐stimulated O2
− generation is not. WGA‐stimulated O2
− generation of macrophages was also inhibited in the presence of dibutyryl cAMP or prostaglandin E2 (PGE2), substances that increase intracellular cAMP, but PMA‐stimulated O2
− generation was only slightly affected by these compounds. These results suggest that the mechanism for O2
− generation of macrophages caused by WGA is different from that for O2
− generation caused by PMA.
Amino acid sequence of a polymorphic segment from fragment C4d of human complement component C4
Authors: Chakravarti D.N. , Campbell R.D. , Gagnon J.
Published: July 2002

The amino acid sequence of a segment of 106 residues of C4d has been determined by automated sequence analysis of fragments obtained by CNBr cleavage and enzymic digestion with trypsin. Polymorphism has been detected at 3 positions. Residues 9 and 12 are either valine and leucine or alanine and arginine, respectively. Residue 102 is either valine or arginine. When comparing the protein sequence with the nucleic, acid sequence [Carroll, M. and Porter, R.R. (1983) Proc. Natl. Acad. Sci. USA 80, in press] alanine or serine are found at position 98. These results may in part help to explain the inherited variants of human C4 seen on gel electrophoresis.
Transcribing chromatin is not preferentially enriched with acetylated histones
Authors: Yukioka Munehiko , Sasaki Satoshi , Henmi Shigeru , Matsuo Mayumi , Hatayama Takumi , Inoue Akira
Published: July 2002

Chromatin fragments of the RNA polymerase II‐transcriptional complex were purified from the micrococcal nuclease digest of rat liver nuclei in the presence of n‐butyrate, a potent histone deacetylase inhibitor. Polyacrylamide gel electrophoretic analysis in Triton acid—urea revealed that the extent of histone acetylation of the complex did not differ markedly from that of the total chromatin.
Interaction of neuroleptic drugs with a synthetic calcium‐binding peptide analog of site III of rabbit skeletal troponin C
Authors: Reid Ronald E. , Gariépy Jean , Hodges Robert S.
Published: July 2002

The effects of nine drugs on the CD spectra of a synthetic calcium binding analog of site III or rabbit skeletal troponin C, can generally be divided into 3 groups: (1) that consisting of haloperidol, benperidol, molindone and promethazine, had no effect on the CD spectrum or calcium sensitivity of the apopeptide; (2) that composed of structurally rigid thioxanthenes, induced CD‐detectable structural change in the apopeptide but prevented Ca2+‐induced structural change; (3) that consisting of chlorpromazine, trifluoperazine and fluphenazine, induced structural change in the peptide but had no effect on the Ca2+‐induced structural change.
Desaturation of oleoyl and linoleoyl residues linked to phospholipids in growing roots of yellow lupin
Authors: Citharel B. , Oursel A. , Mazliak P.
Published: July 2002

When growing lupin roots were fed with [14C]oleate and [14C]linoleate most of the absorbed molecules remained in the free fatty acid pool. Part of the 14C‐labelled fatty acid was integrated first into phosphatidylcholine and subsequently into phosphatidylethanolamine. Then, progressive desaturation took place. The kinetics of these processes suggests that desaturation in phosphatidylcholine was followed by transacylation of polyunsaturated fatty acids into phosphatidylethanolamine. To produce good yields of desaturation (about 48%), seeds had to be less than 1‐year old. With respect to linoleate desaturation, seedlings had to be 2–7‐days old. Increased percentages of oleate desaturation were obtained when the roots were grown on a medium enriched with calcium.
Evolution and structure of two ADP‐ribosylation enterotoxins, Escherichia coli heat‐labile toxin and cholera toxin
Authors: Yamamoto Tatsuo , Nakazawa Teruko , Miyata Takashi , Kaji Akira , Yokota Takeshi
Published: July 2002

Nucleotide sequence comparisons of the heat‐labile enterotoxin (LTh) genes of E. coli pathogenic for humans with cholera toxin (CT) genes suggest that the two toxin genes have evolved from a common ancestry by a series of single selective base changes, while conserving the catalytic fragment A1 (ADP‐ribose transferase). Based on the local hydrophilicity profiles of LTh and CT peptides, a transmembrane segment appears to be present in A1 in both toxins.
Lipid peroxidation initiated by superoxide‐dependent hydroxyl radicals using complexed iron and hydrogen peroxide
Authors: Gutteridge John M.C.
Published: July 2002

Iron salts stimulate lipid peroxidation by decomposing lipid peroxides to produce alkoxyl and peroxyl radicals which initiate further oxidation. In aqueous solution ferrous salts produce OH. radicals, a reactive species able to abstract hydrogen atoms from unsaturated fatty acids, and so can initiate lipid peroxidation. When iron salts are added to lipids, containing variable amounts of lipid peroxide, the former reaction is favoured and OH. radicals contribute little to the observed rate of peroxidation. When iron is complexed with EDTA, however, lipid peroxide decomposition is prevented, but the complex reacts with hydrogen peroxide to form OH. radicals which are seen to initiate lipid peroxidation. Superoxide radicals appear to play an important part in reducing the iron complex.
Evidence for similar structural changes on binding of platinum anti‐tumor agents to DNA and nucleosomes
Authors: Marzilli Luigi G. , Reily Michael D. , Heyl Barbara L. , McMurray Cynthia T. , Wilson W.David
Published: July 2002

The influence of Pt(II) compounds on the 31P NMR spectra of natural DNA, synthetic polynucleotides, and nucleosomes was investigated. With Pt complexes which are anti‐tumor agents, a new peak or shoulder centered at ~ 1.2 ppm downfield from the untreated DNA signal was observed. When Pt compounds known not to be anti‐tumor agents were studied, no such new signal was observed. The most reasonable explanation for the downfield resonance is that it is a consequence of a structural change induced in the DNA by the anti‐tumor agent. Since the effect of the Pt compounds on nucleosomes was similar, the same structural change is probably occurring in DNA in solution and in nucleosomes. A nonalternating dG · dC polymer, but not alternating G·C or any A·T polymers, exhibited a similar spectral change and this finding suggests that the structural change in the DNA arises primarily from reaction of Pt anti‐tumor agents with adjacent G residues.
Development of the 33‐, 23‐ and 16‐kDa polypeptides of the photosynthetic oxygen‐evolving system during greening
Authors: Ryrie Ivan J. , Young Susan , Andersson Bertil
Published: July 2002

Thylakoid development
Photosynthetic oxygen evolution
33‐kDa protein
23‐kDa protein
16‐kDa protein
Western blotting
High enkephalyl peptide degradation, due to angiotensin‐converting enzyme‐like activity in human CSF
Authors: Lantz I. , Terenius L.
Published: July 2002

The metabolism of enkephalin peptides was studied in human cerebrospinal fluid. The degradation rates of (Leu)‐enkephalin and (Leu)‐enkephalin‐Arg6 were compared and the latter was degraded at a 10‐fold higher rate. The major enzyme activity was investigated by M
r determination and inhibition experiments, showing marked similarity with angiotensin‐converting enzyme.
Synthesis and functional activity of translation initiation regions in mRNA
Authors: Renhof R. , Cielens I. , Nikitina T. , Sherinya L. , Shomshtein Z. , Gren E.J.
Published: July 2002

Polyribonucleotide
Enzymatic synthesis
Protein synthesis initiation
Template activity
The synthesis of guanosine 5'‐diphosphate D‐glucose by enzyme extracts of mung beans (Phaseolus aureus) and other higher plants
Authors: Barber G.A.
Published: July 2002

GDP‐D‐glucose
Phaseolus aureus
β‐1,4‐Glucan
The activation of cellulases from different sources by actin
Authors: Klyosov Anatole A. , Bolobova Alla V. , Ostrovskaya Marina V. , Poglazov Boris F.
Published: July 2002

Actin has been shown to be a potent activator of the enzymatic hydrolysis of cellulose, its addition to the reaction mixture leading to an increase in the hydrolysis rate of up to 6–7 fold. The action of actin is directed primarily towards endoglucanases of cellulase complexes. The degree of activation varies in relation to the cellulases from different microbial sources. The activation of the enzymatic hydrolysis of an insoluble cellulose by actin does not affect the Michaelis constant but increases the maximum velocity of the reaction. Increasing the actin concentration leads to a linear increase in the activation effect which indicates a rather poor binding of actin with the cellulases under study.
Membrane protein‐lipid hydrogen bonding: evidence from protein kinase C, diglyceride, and tumor promotors
Authors: Brockerhoff Hans
Published: July 2002

Membrane‐bound proteins owe their retention and conformation in the lipid bilayer to hydrophobic peptide domains. Additional fixation, by protein‐lipid hydrogen bonding, has been suggested, and recent reports on protein kinase C activation by diacylglycerol (DG) provide an unambiguous model for such bonding. The sn‐1,2‐diacylglycerol appears to donate a hydrogen bond from the sn‐3 hydroxyl to the enzyme and to receive two hydrogen bonds, in the sn‐1 and sn‐2 ester CO groups, from the enzyme. This arrangement is confirmed in phorbol ester, a competitive inhibitor of DG for the kinase. This tumor promotor has a nearly identical spatial arrangement of hydrogen bond donor (9α‐OH) and acceptors (12 and 13 ester CO); so have two other tumor promotors, teleocidin and aplysiatoxin. There are reasons to believe that protein kinase C is not the only protein that is bound to membrane lipids by hydrogen bonding, and such bonding will have to be considered in membrane‐associated events such as fusion, cross‐membrane transport, or anesthesia.
Characteristic anticodon sequences of major tRNA species from an extreme thermophile, Thermus thermophilus HB8
Authors: Hara-Yokoyama Miki , Yokoyama Shigeyuki , Watanabe Tatsuo , Watanabe Kimitsuna , Kitazume Masako , Mitamura Yasue , Morii Toshie , Takahashi Seizo , Kuchino Yoshiyuki , Nishimura Susumu , Miyazawa Tatsuo
Published: July 2002

Major species of tRNAGlu, tRNALys, tRNAVal, tRNAAla, tRNASer and tRNAArg were purified from Thermus thermophilus HB8, and their anticodon sequences were found to be CUC, CUU, CAC, GGC, GGA and CCG, respectively, with unmodified G or C as the first letter. These anticodons exactly correspond to the codons as used most frequently among synonymous codons in the structural genes of extreme thermophiles. The stable complexes of these anticodons (in particular, GGN or CCN) with their corresponding codons probably contribute to the correct protein biosynthesis in extreme thermophiles at high temperature.
Patterns of gene expression during plasmacytoid differentiation of chronic lymphocytic leukaemia cells
Authors: Norton J.D. , Leber B. , Yaxley J.C.
Published: July 2002

Chronic lymphocytic leukaemia (CLL) cells may be induced to undergo plasmacytoid differentiation in vitro in response to 12‐O‐tetradecanoyl phorbol acetate (TPA). We show here that plasmacytoid differentiation and the accompanying accumulation of Cμ immunoglobulin mRNA are preceded by a rapid transient increase in the expression of the proto‐oncogenes, c‐myc and c‐fos. In terminally differentiated cells the level of c‐fos mRNA returned to the original basal level whilst c‐myc expression remained appreciably higher than in undifferentiated CLL cells. These data support a possible role for c‐fos and c‐myc in the programmed chain of events that occur during terminal differentiation of B‐lymphocytes.
Inositol phospholipid turnover and protein kinase C translocation are stimulated by poly(I) · poly(C) in human amnion cells (UAC)
Authors: Premecz György , Markovits Andrea , Bagi György , Farkas Tibor , Földes István
Published: July 2002

Polyinosinic‐polycytidylic acid, a potent inducer of interferon (IFN) production and activator of some IFN‐induced enzymes, inhibits [3H]uridine incorporation into the RNA of vesicular stomatitis virus even in the absence of IFN synthesis, transiently triggers the breakdown of inositol phospholipids and activates the translocation of protein kinase C. Since IFNs also have similar activities these results suggest that IFN induction and IFN function are realised through common biochemical pathways.
The DNase I generated disomal series is coherent to 16N Implications for coiling models of chromatin structure
Authors: Davis S.J. , Burgoyne L.A.
Published: July 2002

The disomal series generated by the digestion of chromatin with DNase I has been followed to its highest orders using Klenow end‐labelling and field‐inversion gel electrophoresis to maximise the resolution of large DNA fragments. The series is coherent to the 16N level and as such is incompatible with the most structurally acceptable coiling models. We propose that this is evidence for the general unsuitability of coiling models and is support for the existence of simple “back‐to‐back” double‐stranded structures within chromatin.
Studies on interaction of 5 S RNA with ribosomal proteins
Authors: Kärgel H.-J. , Stahl J. , Gross B. , Knespel S. , Bielka H. , Saarma M.
Published: July 2002

Proteins of the large ribosomal subunit of rat liver (TP 60) were immobilized by diffusion transfer onto nitrocellulose after two‐dimensional polyacrylamide gel electrophoresis (2‐D PAGE). Incubation of the TP 60 blots with 32P‐labeled 5 S RNA under defined ionic conditions (300 mM KCl, 20 mM MgCl2) resulted in specific binding to a limited set of ribosomal proteins consisting of proteins L3, L4, L6, L13/15 and — to a lesser extent — L7 and L19. Under identical conditions, blots with proteins of the small ribosomal subunit (TP 40) did not bind 5 S RNA.
In vitro processing of rANF7–28‐NH2 by rat kidney homogenates
Authors: Berman Judd M. , Pelton John T. , Cardin Alan D. , Blankenship Dale T. , Hassman C.Fred , M. Chen Teng Man
Published: July 2002

The major rat kidney in vitro degradation products of the rat atrial natriuretic factor analog, H‐Cys7‐Phe‐Gly‐Gly‐Arg‐Ile‐Asp‐Arg‐Ile‐Gly‐Ala‐Gln‐Ser‐Gly‐Leu‐Gly‐Cys‐Asn‐Ser‐Phe‐Arg‐Tyr28‐NH2 (with a Cys27‐Cys23 disulfide bridge), have been identified. The degradation products are the C‐terminal modified compounds rANF7–27, rANF7–26, and rANF7–25.
Effect of unbalanced diets on incorporation of δ‐aminolevulinic acid into cytochrome P‐450
Authors: Amelizad Z. , Narbonne J.F. , Borin C. , Robertson L.W. , Periquet A. , Oesch F.
Published: July 2002

The in vivo syntheses of two liver microsomal cytochromes P‐450 PB3a, P‐450 UT50 [(1987) Eur. J. Biochem., submitted] (M
r 50 000, 52 000) have been estimated by measuring the specific activity 2 h after i.p. administration of δ‐[3H]aminolevulinic acid to male Sprague Dawley rats. The animals were fed either a standard rat chow (5% lard, 22% casein) or unbalanced diets (high lipid, 30% lard or low protein, 6% casein) with or without 50 ppm Phenoclor DP6. The high‐lipid diet supported a more rapid body weight gain but had little impact on cytochrome P‐450 content, expressed either per whole liver or per mg microsomal protein, and on the incorporation of the precursor into cytochrome P‐450. The latter was determined by measuring the radioactivity incorporated into the cytochrome P‐450 fraction, partially purified by afffinity chromatography, as well as into two cytochrome P‐450 isozymes (M
r 50 000 or 52 000) purified by DEAE‐52 cellulose ion‐exchange chromatography. The low‐protein diet, on the other hand, severely depressed body weight gain and cytochrome P‐450 content as well as incorporation of radioactivity, the lower‐M
r cytochrome (M
r 50 000) being particularly affected. However, when a potent inducer, Phenoclor DP6, was added to the low‐protein diet, cytochrome synthesis was restored indicating that the effect was reversible.
Immunological evidence for a Physarum β‐tubulin polypeptide possessing an α‐tubulin‐like carboxyl terminus
Authors: Gull Keith , Wilcox Marianne , Birkett Christopher R.
Published: July 2002

A number of well‐characterised monoclonal antibodies have been produced which recognise either α‐ or β‐tubulin. One of these antibodies, YL1/2, has been regarded as a specific probe for α‐tubulin in that it recognises the linear amino acid sequence of the extreme C‐terminus of the α‐tubulin polypeptide. We now report that in the slime mould Physarum polycephalum, YL1/2 recognises the α‐tubulin isotype (α1) that is expressed in the myxamoeba, but that it discriminates between the α1‐tubulin sub‐types of the plasmodium ‐ the other major cell type in the Physarum life cycle; furthermore, YL1/2 does not recognise the plasmodium‐specific α‐tubulin isotype (α1). Intriguingly, with plasmodial tubulin YL1/2 preferentially recognises a β‐tubulin, the plasmodium‐specific β2 isotype. Reactivity with this one specific member of the β‐tubulin family is abolished by treatment of the tubulin with carboxypeptidase. Thus, the plasmodium‐specific β2‐tubulin of Physarum appears to be a β‐tubulin with an α‐tubulin‐like carboxyl terminus.
The secretion of glucagon by transformed yeast strains
Authors: Moody A.J. , Norris F. , Norris K. , Hansen M.T. , Thim L.
Published: July 2002

Saccharomyces cerevisiae strains were transformed with plasmids coding for modified mating factor α1 leader sequences followed by glucagon. Glucagon‐containing peptides which were secreted into the fermentation broth were isolated and their amino acid sequences determined. The yeast strain transformed with the sequence coding for the complete mating factor α1 leader sequence preceding the glucagon gene (MT556) secreted glucagon plus glucagon extended at its N‐terminal by parts of the leader sequence. The yeast strain transformed with the sequence coding for a truncated mating factor α1 leader sequence before the glucagon gene (MT615) secreted glucagon. These observations suggest that S. cerevisiae is a suitable vehicle for the efficient expression of plasmids coding for polypeptides similar to glucagon (e.g. VIP, secretin, GIP).
Interaction of myelin basic protein with the different components of the ATP, Mg‐dependent protein phosphatase system
Authors: Vandenheede Jackie R. , Van Lint Johan , Vanden Abeele Carline , Merlevede Wilfried
Published: July 2002

Myelin basic protein (MBP) reduces the amount of phosphatase activity produced in the kinase FA‐mediated activation of the ATP,Mg‐dependent phosphatase. MBP was shown not only to inhibit the activated enzyme, but also to impair the kinase FA‐mediated activation of the inactive phosphatase. In addition MBP prevents the time‐dependent inactivation of the catalytic subunit by the modulator protein. These observations point to a regulatory role for MBP in the reversible activation of the ATP, Mg‐dependent protein phosphatase by kinase FA.
Chemical probing of the homopurine·homopyrimidine tract in supercoiled DNA at single‐nucleotide resolution
Authors: Vojtíšková M. , Mirkin S. , Lyamichev V. , Voloshin O. , Frank-Kamenetskii M. , Paleček E.
Published: July 2002

Local structure of the homopurine·homopyrimidine tract in a supercoiled plasmid pEJ4 was studied using chemical probes at single‐nucleotide resolution. The conformation of the homopyrimidine strand was probed by osmium tetroxide, pyridine (Os,py) while that of the homopurine strand was tested by diethyl pyrocarbonate (DEPC), i.e. by probes reacting preferentially with single‐stranded DNA. At weakly acidic pH values, a strong Os,py attack on three nucleotides at the centre of the (dC‐dT)16 block and a weaker attack on two nucleotides at the end of the block were observed. DEPC modified adenines in the 5′‐half of the homopurine strand. Os,py modification at the centre of the block corresponded to the loop of the hairpin formed by the homopyrimidine tract, while DEPC modification corresponded to the unstructured half of the homopurine strand in the model of protonated triplex H form of DNA.
Processed enzymatically active protease (p15gag) of avian retrovirus obtained in an E. coli system expressing a recombinant precursor (Pr25lac‐Δgag)
Authors: Sedláček Juraj , Štrop Petr , Kaprálek František , Pečenka Vladimír , Kostka Vladimír , Trávníček Miloslav , Říman Josef
Published: July 2002

Processing proteases of avian and mammalian retroviruses cut the polyprotein precursors encoded by the retroviral genes into mature functional proteins. Retroviral processing proteases are still a rather poorly characterized group as to their relation to other proteases, specificity, and mechanism of enzymatic action. In avian retroviruses the generation of the processing protease itself comprises a processing cleavage event — the protease p15
gag
is cut off the carboxy‐terminus of a gag polyprotein precursor, Pr76
gag
. We report here that direct and efficient production of the avian retrovirus processing protease p15
gag
(required for structure‐function studies and rational design of inhibitors) was obtained in an E. coli system, where massive expression of a size‐reduced, recombinant precursor (Pr25
lac‐Δgag
was accompanied by its structurally accurate processing.
Stereospecific bioactions of 5‐hydroxyicosatetraenoate
Authors: Rossi Adriano G. , Thomas Michael J. , O'Flaherty Joseph T.
Published: July 2002

5(S)‐Hydroxyicosatetraenoate stimulates human polymorphonuclear neutrophils (PMNs) to raise their cytosolic calcium. It also potentiates the cells' degranulation responses to platelet‐activating factor and diacylglycerols. We synthesized 5(R)‐hydroxyicosatetraenoate and found it to be 20–100‐fold weaker than the natural isomer in these assays. Thus, the arachidonic acid metabolite activates PMNs by a stereospecific possibly receptor‐mediated mechanism.
VLDL apoprotein secretion and apo‐B mRNA level in primary culture of cholesterol‐loaded rabbit hepatocytes
Authors: Kosykh V.A. , Surguchov A.P. , Podres E.A. , Novikov D.K. , Sudarickov A.B. , Berestetskaya Yu.V. , Repin V.S. , Smirnov V.N.
Published: July 2002

Incubation of cultured rabbit hepatocytes with β very low density lipoproteins (β‐VLDL) induces a dose‐dependent increase in cell cholesterol (CH) content and VLDL apoprotein (apo) B and E secretion without change in apo‐B mRNA level. These data suggest that β‐VLDL may exert a stimulatory effect on hepatic apo‐B production at the co‐translational and/or posttranslational level.
Effect of chemical modification at C1 of the glucosamine backbone of lipid A‐subunit analog GLA‐27 on manifestation of immunopharmacological activity
Authors: Kumazawa Yoshio , Takimoto Hiroaki , Yamamoto Akihiro , Homma J.Yuzuru , Ogawa Yuji , Kiso Makoto , Hasegawa Akira
Published: July 2002

The effect of chemical modification at the C1 position of GLA‐27, 4‐O‐phosphono‐D‐glucosamine carrying N‐3‐tetradecanoyloxytetradecanoyl [C14‐O‐(C14)] and 3‐O‐tetradecanoyl (C14) groups, was investigated. Replacement by SH or S‐acetyl groups of the OH group resulted in the enhancement of mitogenicity but gave rise to a reduction, in macrophage‐stimulating ability such as induction of tumor necrosis factor and enhancement of phagocytic and cellular acid phosphatase activities. Bisphosphorylation at C1 and C4 resulted in a slight decrease in mitogenicity or almost complete loss of the macrophage‐stimulating ability.
Physiological roles of animal succinate thiokinases Specific association of the guanine nucleotide‐linked enzyme with haem biosynthesis
Authors: Jenkins T.M. , Weitzman P.D.J.
Published: July 2002

The discovery of two distinct succinate thiokinases in mammalian tissues, one (G‐STK) specific for GDP/GTP and the other (A‐STK) for ADP/ATP, poses the question of their differential metabolic roles. Evidence has suggested that the A‐STK functions in the citric acid cycle in the direction of succinyl‐CoA breakdown (and ATP formation) whereas one role of the G‐STK appears to be the re‐cycling of succinate to succinyl‐CoA (at the expense of GTP) for the purpose of ketone body activation. A third metabolic participation of succinyl‐CoA is in haem biosynthesis. This communication shows that in chemically induced hepatic porphyria, when the demand for succinyl‐CoA is increased, it is the level of G‐STK only which is elevated, that of A‐STK being unaffected. The results implicate G‐STK in the provision of succinyl‐CoA for haem biosynthesis, a conclusion which is further supported by the observation of a high G‐STK/A‐STK ratio in bone marrow.
The activation and inactivation of the Dunaliella salina chloroplast coupling factor 1 (CF1) in vivo and in situ
Authors: Selman-Reimer Susanne , Selman Bruce R.
Published: July 2002

Preillumination of intact cells of the eukaryotic, halotolerant, cell‐wall‐less green alga Dunaliella salina induces a dark ATPase activity the magnitude of which is about 3–5‐fold higher than the ATPase activity observed in dark‐adapted cells. The light‐induced activity arises from the activation and stabilization in vivo of chloroplast coupling factor 1 (CF1). This activity, ∼ 150–300 μmol ATP hydrolyzed/mg Chl per h, rapidly decays (with a half‐time of about 6 min at room temperature) in intact cells but only slowly decays (with a half‐time of about 45 min at room temperature) if the cells are lysed by osmotic shock immediately after illumination. The activated form of the ATPase in lysed cells is inhibited if the membranes are treated with ferri‐ but not ferrocyanide, suggesting that the stabilization of the activated form of CF1 is due to the reduction of the enzyme in vivo in the light.
The terminal protein of minute virus of mice is an 83 kilodalton polypeptide linked to specific forms of double‐stranded and single‐stranded viral DNA
Authors: Gunther Magali , Tattersall Peter
Published: July 2002

A new assay (label transfer from DNA to protein) enabled the identification of a terminal protein (TP) in nucleoprotein complexes extracted from cells infected with the parvovirus, minute virus of mice, MVM. In SDS‐PAGE, TP migrates as a major band at 83 kDa, with a minor 65 kDa component, each of which exactly co‐migrates with the cellular forms of the virally coded polypeptide NS‐1. In parallel, the analysis of nucleoproteins by SDS‐agarose gel electrophoresis allowed us to observe that the major species of viral DNA molecules (mRF, dRF and ssDNA) are all present in the form of DNA‐protein complexes. Three forms of mRF DNA were identified, two of which are protein‐associated and one which appears to be protein‐free.
A liver‐specific nuclear protein that binds to the distal promoter element of the rat tyrosine aminotransferase gene
Authors: Lobanenkov V.V. , Klenowa E.M. , Adler V.V.
Published: July 2002

Using a gel retardation assay and exonuclease III footprinting, we have analyzed sequence‐specific DNA‐binding nuclear factors which interact with the distal promoter element of the rat tyrosine aminotransferase gene. A factor called LspA1, binding to a sequence that resembles the consensus binding site for the transcription factor Ap‐1, was shown to be present in adult rat‐liver nuclear protein extracts but not in the extracts from embryonic liver or spleen nuclei.
Monoclonal antibodies against the S2‐serotonin receptor from rat brain that cross‐react with dopamine and opiate receptors
Authors: Deckmann M. , Shinitzky M.
Published: July 2002

Balb/c mice were immunized with rat striatal integral membrane proteins. After hybridization of splenocytes with myeloma cells, hybridoma lines secreting antibodies against serotonin, dopamine and opiate receptors were detected by inhibition of ligand binding to brain membrane preparations. Antibodies from two positive lines, Mab/a9 and Mab/a18, were able to inhibit ligand binding to the S2‐serotonin (K
d range: 10–100 nM), the μ‐opiate (K
d range: 0.4–3 μM) and the δ‐opiate receptors (K
d range: 0.7–1.1 μM), while Mab/a9 was also found to inhibit ligand binding to the D2/D4‐dopamine receptor (K
d ∼ 50 nM). An apparent molecular mass of 60 kDa could be ascribed to the δ‐opiate receptor and apparent molecular masses of 29 and 36 kDa to the μ‐opiate receptor by ligand elution from immuno‐precipitates.
Identification of cholera toxin‐binding sites in the nucleus of intestinal epithelial cells
Authors: Parkinson Marianne E. , Smith Colin G. , Garland Peter B. , van Heyningen Simon
Published: July 2002

Post‐embedding immunogold electron microscopy shows several binding sites for cholera toxin in mouse intestinal epithelial cells, particularly in the heterochromatin of the nucleus as well as in the plasma membrane. Anti‐ganglioside GM1 antibodies also bound to the nucleus, but did not interfere with the binding of toxin. 125I‐labelled toxin bound specifically to a nuclear preparation from rabbit intestinal cells.
Hydroxyl radical generation and DNA strand scission mediated by natural anticancer and synthetic quinones
Authors: Rumyantseva Galina V. , Weiner Lev M. , Frolova Elena I. , Fedorova Olga S.
Published: July 2002

Using ESR and spin‐trapping techniques, it was found that synthetic 2‐dimethylamino‐3‐chloro‐1,4‐naphthoquinone and the natural anticancer quinone daunomycin, when added to a system containing purified NADPH‐cytochrome P‐450 reductase, NADPH, ferric ions, and oxygen, (i) generated hydroxyl radicals and (ii) caused single‐strand scission of super‐coiled DNA of the plasmic pBR322. Since these two effects of the quinones were correlated to each other, we propose that potential anticancer quinones can be effectively screened by measuring their ability to form hydroxyl radicals in the above system.
Expression of high‐ and low‐affinity epidermal growth factor receptors in human hepatoma cell lines
Authors: Clementi Massimo , Festa Antonella , Testa Ivano , Bagnarelli Patrizia , Devescovi Giulia , Carloni Guido
Published: July 2002

Data are presented from a comparative research on expression of epidermal growth factor (EGF) receptors and response to EGF of six independently established cell lines derived from human hepatoma. These lines differ in terms of the degree of differentiation, presence of hepatitis B virus (HBV) DNA copies in integrated form and expression of HBV genes. Our results indicate differential expression of membrane EGF receptors and differential response to EGF under serum‐ and hormone‐free culture conditions. Furthermore, a significant difference in affinity could be detected between EGF receptors of the two highly dedifferentiated cell lines (HA22T/VGH and Li7A) whose replication is inhibited by EGF concentrations capable of stimulating more differentiated phenotypes.
The complete islet amyloid polypeptide precursor is encoded by two exons
Authors: Mosselman S. , Höppener J.W.M. , Lips C.J.M. , Jansz H.S.
Published: July 2002

Islet amyloid polypeptide (IAPP) is the 37‐amino acid peptide subunit of amyloid found in pancreatic islets of type 2 diabetic patients and in insulinomas. Recently, we isolated the human gene encoding IAPP [(1988) FEBS Lett. 239, 227–232]. We now report the nucleotide sequences of a human insulinoma cDNA encoding a complete IAPP precursor, and of the corresponding parts of the IAPP gene. Two exons, which are approx. 5 kb apart in the human genome, encode the 89‐amino acid pre‐pro‐IAPP. At least one additional exon is present further upstream in the IAPP gene. A putative signal sequence at the amino‐terminus of the precursor suggests that IAPP is a secreted protein.
Caldesmon150, caldesmon77 and skeletal muscle troponin T share a common antigenic determinant
Authors: Shirinsky Vladimir P. , Biryukov Konstantin G. , Vorotnikov Alexander V. , Gusev Nikolai B.
Published: July 2002

Caldesmon and troponin are known to control actin‐myosin interaction in smooth and striated muscle. We showed that polyclonal antibodies to smooth muscle caldesmon150 cross‐react with non‐muscle caldesmon77 and with skeletal muscle troponin T. The epitope recognized by caldesmon antibodies is located in the CB 2 peptide [residues 71–151] of troponin T. It is supposed that the observed cross‐reactivity is due to the common structure of the tropomyosin‐binding site of caldesmon and troponin T.
Heterogeneity within the alkane‐inducible cytochrome P450 gene family of the yeast Candida tropicalis
Authors: Sanglard Dominique , Fiechter Armin
Published: July 2002

Alkane; Cytochrome P450 gene family; Yeast; (Candida tropicalis)
Isolation and sequence determination of human brain natriuretic peptide in human atrium
Authors: Kambayashi Yoshikazu , Nakao Kazuwa , Mukoyama Masashi , Saito Yoshihiko , Ogawa Yoshihiro , Shiono Shozo , Inouye Ken , Yoshida Nobuo , Imura Hiroo
Published: July 2002

We isolated human brain natriuretic peptide (human BNP) from the human atrium. Sequence analysis has revealed that it is a 32‐amino‐acid peptide with the sequence S‐P‐K‐M‐V‐Q‐G‐S‐G‐C‐F‐G‐R‐K‐M‐D‐R‐I‐S‐S‐S‐S‐G‐L‐G‐C‐K‐V‐L‐R‐R‐H, which is identical to the C‐terminal sequence (77–108) of the human BNP precursor deduced from the cDNA sequence. The sequence of human BNP (77–108) is preceded by Pro75‐Arg76 in the human BNP precursor, which is the same processing signal as Pro97‐Arg98 of the precursor of atrial natriuretic peptide (ANP). The processing of the BNP precursor occurs in the cardiocyte, although that of the ANP precursor in the cardiocyte is unclear at present.
Effect of polyelectrolytes on serine proteinase secretion by Bacillus subtilis
Authors: Artemov Anatoly V. , Samuilov Vitaly D.
Published: July 2002

Addition of polycations with molecular masses of 5–40 kDa as well as Na+, stimulated serine proteinase secretion by Bacillus subtilis cells. Polyanions and higher‐molecular‐mass polycations (100–200 kDa) were inefficient. The enzyme yields in the presence of polycations or Na+ were equal in magnitude. The results indicate that the cations, apparently counteracting the negative surface charge of the bacterial plasma membrane, cause the desorption of the serine (alkaline) proteinase. The synthesis of the proteinase is inferred to be stopped as the enzyme is bound to the outer surface of the plasma membrane. The desorption of the enzyme thus induces the synthesis of the new portions of proteinase.
Expression of subunit III ot the ATP synthase from spinach chloroplasts in Escherichia coli
Authors: Burkovski Andreas , Deckers-Hebestreit Gabriele , Altendorf Karlheinz
Published: July 2002

Expression of subunit III of the ATP synthase from spinach chloroplasts in Escherichia coli has been achieved. Although the protein is inserted into the bacterial cytoplasmic membrane, formation of a functional Fo complex was not observed.
Specific binding of sulphated polymers to ram sperm proacrosin
Authors: Williams Rachel M. , Jones Roy
Published: July 2002

Sulphated polysaccharides and zona pellucida glycoproteins have been shown to bind non‐enzymatically to proacrosin, the protein found within the acrosomal vesicle of mammalian spermatozoa. The mechanism of this interaction has been investigated using 125I‐fucoidan to probe purified ram sperm proacrosin. Results show that (a) binding of' 125I‐fucoidan to proacrosin is inhibited only by sulphated polymers and (b) recognition is mediated by poly(sulphate) groups and is largely independent of the composition of the polymer chain. It is suggested that a similar mechanism is responsible for the interaction between proacrosin and zona pellueida glycoproteins during the early stages of fertilization in mammals and this process mediates firm binding of spermatozoa to the egg.
Emerging views on the structure and dynamics of the Ca2+‐ATPase in sarcoplasmic reticulum
Authors: Martonosi Anthony N. , Jona Istvan , Molnar Elek , Seidler Norbert W. , Buchet Rene , Varga Sandor
Published: July 2002

The ATP‐dependent Ca2+ transport in sarcoplasmic reticulum involves transitions between several structural states of the Ca2+‐ATPase, that occur without major changes in the secondary structure. The rates of these transitions are modulated by the lipid environment and by interactions between ATPase molecules. Although the Ca2+‐ATPase restricts the rotational mobility of a population of lipids, there is no evidence for specific interaction of the Ca2+‐ATPase with phospholipids. Fluorescence polarization and energy transfer (FET) studies, using site specific fluorescent indicators, combined with crystallographic, immunological and chemical modification data, yielded a structural model of Ca2+‐ATPase in which the binding sites of Ca2+ and ATP are tentatively identified. The temperature dependence of FET between fluorophores attached to different regions of the ATPase indicates the existence of ‘rigid’ and ‘flexible’ regions within the molecule characterized, by different degrees of thermally induced structural fluctuations.
Nitrogen fixation genes involved in the Bradyrhizobium japonicum‐soybean symbiosis
Authors: Hennecke Hauke
Published: July 2002

The symbiotic nitrogen fixation genes (nif, fix) of Bradyrhizobium japonicum, the root nodule endosymbiont of soybean, are organized in at least two separate chromosomal gene clusters. These genes code for proteins of the nitrogenase complex, for proteins involved in their assembly with cofactors and for putative electron transport functions. One gene, nif A, codes for a transcriptional regulatory protein that plays a central role in the control of expression of the other genes in response to the cellular oxygen status. Only at low partial pressures of O2 will the target promoters be activated by NifA.
Osmium tetroxide, N,N,N',N'‐tetramethylethylendiamine A new probe of DNA structure in the cell
Authors: Boublíková P. , Paleček E.
Published: July 2002

It was shown that the complex of osmium tetroxide with N,N,N',N'‐tetramethylethylendiamine can be applied as a probe of DNA structure in the cell. This probe site‐specifically recognized structural distortions at the B‐Z junctions in plasmids pRW751 and pPK1 (containing (dC‐dG)
n
. segments) in E. coli cells.
Expression of a truncated protein‐tyrosine phosphatase mRNA in human lung
Authors: Vries Luc De , Li Ruo-Ya , Ragab Ashraf , Ragab-Thomas Jeannie M.F. , Chap Hugues
Published: July 2002

Protein‐tyrosine phosphatases (PTPases) are becoming an important family of enzymes that might regulate key events in cell growth and transformation. While isolating a new member of this family via amplification of human lung cDNA by the polymerase chain reaction, we found a clone identical to but truncated at the 3′‐end of the coding region of human PTPase β (HPTPβ) mRNA. This difference in sequence is situated in the most conserved part of the catalytic domain of the enzyme. The expression level of the truncated form or HPTPβ mRNA in human lung was lower than its normal form