bluuebunny/crossref_metadata_2025 huggingface dataset:
Mapping of two tyrosine residues involved in the quinone‐ (QB) binding site of the D‐1 reaction center polypeptide of photosystem II
Authors: Dostatni Ralf , Meyer Helmut E. , Oettmeier Walter
Published: July 2002

The property of the D‐1 subunit of photosystem II in binding herbicides in its quinone‐binding niche has provided important approaches to study its structure and function. In the D‐1 protein, amino acid residues Tyr‐254 and Tyr‐237 are labeled by an [14C]azido‐urea derivative, as identified by protein sequencing of proteolytic fragments. Whereas Tyr‐254 is in a parallel α‐helix already indicated to contribute to the herbicide‐binding site, Tyr‐237 is in a hydrophilic sequence that is partly accessible from the matrix space of the chloroplasts. This area has been implicated to contain a cleavage site for a protease involved in the rapid turnover of the D‐1 polypeptide. The photoaffinity labeling results show that some of the amino acids in this cleavage site are actually part of the quinone‐binding niche. It allows a refined and extended prediction of the three‐dimensional folding of the reaction center of photosystem II.
Integration and loss of a single ν‐Ki‐ras gene affects tumorigenic potential of human osteosarcoma cells
Authors: Carloni Guido , Venuat Anne-Marie , Daya-Grosjean Leela , Nardeux Pierre , Rhim Johng S. , Azzarone Bruno
Published: July 2002

The human osteosarcoma cell line Te85 clone F‐5 is not tumorigenic in vivo. Its transformation with Kirsten murine sarcoma virus (KiMSV) (KHOS) confers full malignant properties and stable non‐tumorigenic revertants of this KHOS cell line have been obtained. Here we show that integration and expression of a single copy of the KiMSV proviral DNA, which is totally lost in the HOS 240S revertant, is responsible for the acquisition of tumorigenicity. Cytogenetic analysis and the absence of a residual LTR copy in the revertant cellular genome suggest that the loss of KiMSV provirus is caused either by chromosomal segregation or by recombination not involving the LTR. In addition analysis of the expression of ras proteins revealed no changes in the pattern of c‐ras products and the expression of v‐ras only in the KHOS cells. All these data suggest that Te85 and HOS 240S cell lines could represent a human alternative recipient system to rodent cells in studies with oncogenes.
Uranyl salts as photochemical agents for cleavage of DNA and probing of protein‐DNA contacts
Authors: Nielsen Peter E. , Jeppesen Claus , Buchardt Ole
Published: July 2002

Single‐strand DNA nicks are induced by uranyl nitrate or uranyl acetate in combination with long‐wavelength (λ ∼ 420 nm) ultraviolet irradiation. The nicks occur randomly with respect to the DNA sequence. Using the λ‐repressor/OR1 operator DNA system it is shown that uranyl salts can be used to photofootprint protein contacts with the DNA backbone.
Photoaffinity labeling of a 33 kDa protein subunit of the δ‐opioid receptor in neuroblastoma and hybrid cell lines
Authors: Icard-Liepkalns C. , Bochet P.
Published: July 2002

Tritiated DTLET (Tyr‐D‐Thr‐Gly‐Phe‐Leu‐Thr) binds with high affinity, specificity and saturability to neuroblastoma N18TG2 and hybrid neuroblastoma × glioma NG108‐15 and NG 108‐5 intact cells. The δ‐opioid receptor density in cells cultured in chemically defined medium was increased about 2 times compared to that in cells cultured in 10% fetal calf serum. A major and a minor protein species covalently and specifically bound to [125I]azido‐DTLET (Tyr‐D‐Thr‐Gly‐pN3Phe‐Leu‐Thr), photoactivatable ligand, migrated on SDS‐gel electrophoresis with M
r, values near 33000 and 58000, respectively.
Complete sequence of the gene encoding the largest subunit of RNA polymerase I of Trypanosoma brucei
Authors: Jess Waldemar , Hammer Andrea , Cornelissen Albert W.C.A.
Published: July 2002

We have set out to clone the trypanosomal gene encoding the largest subunit of RNA polymerase I. We screened a genomic library with a synthetic oligonucleotide probe encoding an eleven amino acid sequence motif, YNADFDGDEMN, which has been found in all eukaryotic RNA polymerase largest subunit genes analyzed so far. We isolated the Trp11 locus and determined the complete sequence of the gene encoded within this locus. The deduced amino acid sequence contains the highly conserved RNA polymerase domains as well as the previously identified RNA polymerase I‐specific hydrophilic insertions. Therefore, the gene most closely resembles the largest subunit of RNA polymerase I.
Evidence for a protein kinase C‐directed mechanism in the phorbol diester‐induced phospholipase D pathway of diacylglycerol generation from phosphatidylcholine
Authors: Cabot Myles C. , Welsh Clement J. , Zhang Zu-chuan , Cao Hui-ting
Published: July 2002

In this study we provide evidence for the involvement of protein kinase C (PKC) in phorbol diester‐induced phosphatidylcholine (PC) hydrolysis by the phospholipase D pathway. Rat embryo fibroblasts (REF52) were prelabeled with either tritiated choline or myristic acid; these compounds are preferentially incorporated into cellular PC. Phorbol diester‐induced PC degradation was determined by measuring the release of [3H]choline, and the formation of [3H]myristoyl‐containing phosphatidate (PA), diacylglycerol (DG), and phosphatidylethanol (PE). Staurosporine, a PKC inhibitor, blocked from 73 to 90% of the phorbol diester‐induced PC hyrolysis. The inhibition of phorbol diester‐induced choline release by staurosporine was dose dependent with an approximate ED50 of 150 nM. Pretreatment of cells with phorbol diester inhibited subsequent phorbol diester‐induced PC degradation by 78–92%. A close correlation between the ED50 for phorbol diester‐stimulated choline release and the K
d for phorbol diester binding was demonstrated. Neither forskolin nor dibutyryl cAMP elicited cellular PC degradation. In vitro experiments using phospholipase D from Streptomyces chromofuscus showed that staurosporine did not inhibit and TPA did not stimulate enzyme activity.
Oligonucleotide duplexes containing CC(A/T)GG stimulate cleavage of refractory DNA by restriction endonuclease EcoRII
Authors: Pein Claus-Dietmar , Reuter Monika , Cech Dieter , Krüger Detlev H.
Published: July 2002

Some DNA species are resistant towards the restriction endonuclease EcoRII despite the presence of unmodified recognition sites. We show that 14 base‐pair oligonucleotide duplexes containing the EcoRII recognition site 5′‐CC(A/T)GG are cleaved by this enzyme and are able to stimulate EcoRII cleavage of such resistant DNA molecules (e.g. DNA of bacterial virus T3). A direct correlation between the concentration of oligonucleotide duplex molecules and the degree of EcoRII digestion of the primarily resistant DNA is observed. This indicates a stoichiometric rather than a catalytic mode of enzyme activation. An excess of DNA devoid of EcoRII sites (‘non‐site’ DNA, e.g. MvaI‐digested T7 DNA) does not interfere with the activity of EcoRII.
Interleukin 3 and phorbol ester stimulate tyrosine phosphorylation of overlapping substrate proteins
Authors: Ferris Douglas K. , Willette-Brown Jami , Martensen Todd , Farrar William L.
Published: July 2002

FDC‐P1 is a murine myeloid cell line that requires interleukin 3 (IL3) for survival and proliferation. While the biological effects of IL3 have been well described, the biochemical mechanisms of IL3 actions have only recently been examined. We have investigated whether IL3 or PMA stimulates phosphorylation of proteins on tyrosine as well as on serine/threonine residues as previously described [(1986) Blood 68, 906–913; (1987) Biochem. J. 244, 683–691]. Here we report that both IL3 and PMA stimulate the tyrosine phosphorylation of at least two proteins: pp70 and pp50 in FDC‐P1 cells.
The roles of protein kinase C and cyclic nucleotide dependent kinase in signal transduction in human interferon γ induction by poly I:poly C
Authors: Tamura-Nishimura Motoko , Sasakawa Shigeru
Published: July 2002

The signal transduction mechanisms involved in interferon (IFN) γ induction in human peripheral mononuclear lymphocyte nylon‐nonadherent cells (NNA cells) by stimulation with poly(I):poly(C) are investigated. Significant enhancement of IFN γ production by poly(I):poly(C) is observed in the presence of the phorbol ester, 12‐O‐tetradecanoylphorbol 13‐acetate (TPA, a protein kinase C (PKC) activator). Our study shows that in NNA cells, poly(I):poly(C) with or without TPA causes prolonged activation of cytosolic PKC of NNA cells for at least 120 min. The level of activation of PKC is quite remarkable in the case of the combined stimulation by poly(I):poly(C) and TPA as compared to poly(I):poly(C) alone. This demonstrates that prolonged activation of cytosolic PKC for at least 120 min is essential for high levels of production of IFN γ. Moreover, inhibition experiments using the PKC inhibitor H‐7 and cAMP‐dependent protein kinase is inhibitor H‐8 suggest that the mechanism of signal transduction with regard to PKC is involved in stimulation of IFN γ production in NNA cells by poly(I):poly(C) in the presence of TPA and that along with PKC, cAMP‐dependent protein kinase is probably involved in induction of IFN γ by stimulation with poly(I):poly(C) alone.
Eicosanoids are regulatory molecules in γ‐interferon‐induced endothelial antigenicity and adherence for leucocytes
Authors: Leszczynski Dariusz , Häyry Pekka
Published: July 2002

When the endothelial cells (ECs) were stimulated with γ‐interferon (gIFN) in the presence of methylprednisolone (MP) or prostaglandin E2 (PGE2), MP enhanced gIFN‐induced Ia antigen expression, whereas PGE2 inhibited it. On the other hand, while PGE2 had no effect on leucocyte binding to ECs, MP entirely inhibited it. By using selective inhibitors of the cyclo‐oxygenase pathway (indomethacin, IM) and the 5‐lipoxygenase pathway (L651.392), we found that addition of IM to gIFN‐stimulated ECs enhanced Ia expression but had no effect on leucocyte adherence to ECs. Instead, addition of L651.392 to gIFN‐stimulated ECs partially reduced leucocyte adherence to ECs but had no effect on Ia expression. Pretreatment of the ECs or leucocytes or both with monoclonal anti‐class II antibody, had no effect on gIFN‐induced leucocyte binding to ECs. These findings suggest that gIFN‐induced endothelial cell antigenicity and leucocyte adherence are regulated independently of each other by different molecular pathways. Moreover, arachidonic acid metabolites appear to be the regulatory molecules in gIFN effects on the ECs.
Subtype‐specific increase in G‐protein α‐subunit mRNA by interleukin 1β
Authors: Lee Richard T. , Brock Tommy A. , Tolman Cynthia , Bloch Kenneth D. , Seidman J.G. , Neer Eva J.
Published: July 2002

The guanine nucleotide regulatory proteins (G‐proteins) which are substrates for ADP‐ribosylation by pertussis toxin (αi−1, αi−2, αi−3 and α0) transduce a variety of hormonal signals. Endothelial cells express mRNA for three αi subtypes although the level of αi−1 mRNA is very low. Interleukin 1β (IL1β), a pleiotropic inflammatory mediator which stimulates a complex series of responses in human endothelial cells leading to increased coagulation and platelet adhesion, increases expression of one subtype of αi (αi−2) mRNA in human endothelial cells as determined by Northern blot analysis without affecting the level of mRNA for other α‐subunits. These studies show that mRNA levels for αi subtypes are independently regulated, suggesting that there may be subtype specificity in the cell's requirements for the Gi class of signal‐transducing proteins.
Identification of the plastocyanin binding subunit of photosystem I
Authors: Hippler Michael , Ratajczak Rafael , Haehnel Wolfgang
Published: July 2002

Plastocyanin is specifically cross‐linked by incubation with N‐ethyl‐3‐[3‐(dimethylamino)propyl]carbodiimide (EDC) to a subunit of photosystem I in stroma lamellae and in isolated photosystem I complex. SDS‐PAGE shows the disappearance of a 18.5 kDa subunit and the appearance of a new 31.5 kDa protein which was recognized by anti‐plastocyanin antibodies. The isolated subunit was identified by its N‐terminal amino acid sequence as the mature peptide coded by the nuclear gene psaF [Steppuhn et al. (1988) FEBS Lett. 237, 218–224]. P700+ was reduced by cross‐linked plastocyanin with the same halftime of 13 μs as found in the native complex. This is evidence that cross‐linking conserved the orientation of the complex and that the 18.5 kDa subunit provides the conformation of photosystem I necessary for the extremely rapid electron transfer from plastocyanin to P700+.
cDNA‐directed expression of human thyroid peroxidase
Authors: Kimura Shioko , Kotani Tomio , Ohtaki Sachiya , Aoyama Toshifumi
Published: July 2002

A human thyroid peroxidase cDNA, hTPO‐1 [(1987) Proc. Natl. Acad. Sci. USA 84, 5555–5559], was expressed in human Hep G2 cells using a vaccinia virus cDNA‐expression system. When examined by immunoblot analysis, the level of hTPO‐1 protein expression reached a maximum approx. 24 h after infection and remained at a similar level up to 72 h post‐infection. The expressed protein was enzymatically active as measured by guaiacol oxidation. Monoclonal antibody‐assisted immunoaffinity column chromatography was used for partial purification of vaccinia‐expressed hTPO‐1, resulting in more than 300‐fold higher specific activity and a measurable difference spectrum of the hTPO‐1 (Fe3+)‐CN complex.
Cell cycle related change of Ara‐C transport in HL‐60 cells after differentiation induction
Authors: Takimoto Tetsuya , Kubota Masaru , Tsuruta Satoru , Kitoh Toshiyuki , Tanizawa Akihiko , Akiyama Yuichi , Mikawa Haruki
Published: July 2002

Using a promyelocytic leukemia cell line, HL‐60, we studied the membrane transport of Ara‐C before and after differentiation induced by retinoic acid (RA). In RA‐treated cells, Ara‐C transport was reduced and there was a concomitant increase of the ID50 values of Ara‐C in comparison with the controls. By three different procedures to synchronize untreated cells, i.e. density arrest G1 phase enrichment, aphidicolin‐induced S phase accumulation and the double isoleucine block method, we found that Ara‐C transport was 30–50% higher in the S phase than in the G1 phase. Therefore, the observed decrease in Ara‐C transport is, in part, due to the retarded growth accompanied by an accumulation of cells in the G1 phase after differentiation induction.
A cross‐linked complex between horse pancreatic lipase and colipase
Authors: Chaillan Catherine , Rogalska Ewa , Chapus Catherine , Lombardo Dominique
Published: July 2002

The water soluble carbodiimide N‐cyclohexyl‐N'‐2‐morpholinoethyl‐carbodiimide‐methyl‐p‐toluolsulfonate was found to effectively covalently cross‐link pancreatic colipase to lipase as evidenced by Western blotting experiments using antibodies directed either against lipase or colipase. Moreover the resulting covalent complex has a M
r, consistent with a stoichiometry of 1 mol colipase per mol lipase. Cross‐linked lipase and colipase retain their activity implying a correct covalent binding between the two proteins. The specificity of the lipase‐colipase binding was further supported by the very low amount of cross‐linked products when lipase or colipase alone were incubated in the presence of carbodiimide. The formation of a covalent lipase‐colipase complex in the presence of carbodiimide clearly demonstrates that the binding between both proteins involves ion pairing. Furthermore, the formation of an active covalent complex strongly suggests that the lipase‐colipase binding site is distinct from the colipase interfacial recognition site as well as from the lipase catalytic site.
Presence of paf‐acether in human thymus
Authors: Salem Patrick , Denizot Yves , Pitton Christina , Dulioust Anne , Bossant Marie-Jeanne , Benveniste Jacques , Thomas Yolene
Published: July 2002

Platelet‐activating factor‐acether; Thymus
The formyl peptide chemoattractant receptor is encoded by a 2 kilobase messenger RNA
Authors: Murphy Philip M. , Gallin Elaine K. , Tiffany H.Lee , Malech Harry L.
Published: July 2002

Activation of the formyl peptide chemoattractant receptor (FPCR) of phagocytic cells mobilizes intracellular calcium stores and affects the plasma membrane potential. Affinity crosslinking of FPCR has demonstrated a 60–80 kDa glycoprotein, with core peptide of 32 kDa. It is not known whether functional FPCR is this single peptide or requires multiple subunits. We used Xenopus oocyte expression system to determine the size of mRNA required for synthesis of functional FPCR. Injection of oocytes with poly(A)+ RNA from HL60 cells differentiated to the granulocyte phenotype resulted in acquisition of formyl peptide‐specific responses (inward transmembrane current with a reversal potential consistent with a chloride conductance, and calcium efflux). FPCR activity expressed in oocytes had a ligand concentration dependence, ligand structure dependence and pertussis toxin sensitivity similar to those reported in phagocytic cells. When RNA was size fractionated, a single peak of FPCR activity at 2 kilobases was observed after injection of mRNA into oocytes. Our data strongly suggest that FPCR is composed of a single‐sized polypeptide.
Inhibition of oxygen toxicity by targeting Superoxide dismutase to endothelial cell surface
Authors: Inoue Masayasu , Watanabe Nobukazu , Morino Yoshimasa , Tanaka Yoshikazu , Amachi Teruo , Sasaki Junzo
Published: July 2002

Since enzymes that degrade reactive oxygens, such as Superoxide dismutase (SOD), are significantly lower in plasma than in intracellular compartments, cell surface membranes should be protected against hazardous oxygens particularly when animals are challenged with oxidative stress. To minimize oxygen toxicity on endothelial cell surface, a fusion gene consisting of cDNA coding human Cu2+/Zn2+‐SOD and heparin‐binding peptide was constructed and expressed in yeast. The resulting enzyme (HB‐SOD) bound to a heparin‐Sepharose column and cultured endothelial cells; binding was inhibited either by high NaCl concentrations or heparin. When injected intravenously, HB‐SOD predominantly bound to vascular endothelial cell surface. Carrageenin‐induced paw edema and cold‐induced brain edema of the rat were markedly inhibited by a single dose of HB‐SOD. These results suggest that Superoxide radical and/or its metabolite(s) occurring at or near the outer surface of vascular endothelial cells might play a critical role in the pathogenesis of vasogenic edema.
Resonance Raman spectra of large pea phytochrome at ambient temperature
Authors: Tokutomi Satoru , Mizutani Yasuhisa , Anni Helen , Kitagawa Teizo
Published: July 2002

Resonance Raman (RR) scattering from large pea phytochrome was observed at ambient temperature for the first time by using a micro‐spinning cell and the two color excitation technique. The relative population of the red‐absorbing form (Pr), the far red‐absorbing form (Pfr), and the bleached intermediate (Ibl) under laser illumination was estimated from the absorption spectra. The RR spectrum of Pr obtained by the 363.8 nm excitation under the 740.0 nm pumping exhibited a frequency shift between the H2O and D2O solutions, but those of Pfr and Ibl obtained by the 406.7 nm excitation under the 632.8 nm pumping did not, indicating a distinct difference in their protonation levels, presumably in the protonation of ring C.
Dephosphin, a 96 000 Da substrate of protein kinase C in synaptosomal cytosol, is phosphorylated in intact synaptosomes
Authors: Robinson Phillip J.
Published: July 2002

A 96 000 dalton phosphoprotein, called dephosphin, is phosphorylated in intact synaptosomes from rat brain and is rapidly dephosphorylated upon depolarisation‐dependent calcium entry. A 96 000 dalton phosphoprotein is also a substrate of protein kinase C synaptosomal cytosol, and the aim of the study was to determine whether the two proteins may be the same. Dephosphin in intact synaptosomes and the 96 000 dalton protein kinase C substrate comigrated on polyacrylamide gels. Both phosphoproteins had identical phosphopeptide maps after digestion with V8 protease. Both phosphoproteins ran on isoelectric focussing gels with a pf of 6.3–6.7 and focussed as a series of 5–6 spots. Both proteins were phosphorylated exclusively on serine. Both proteins could be resolved into a doublet on longer polyacrylamide gels. The two subunits were of 96 and 93 kDa in both phosphorylation conditions and had dissimilar phosphopeptide maps. However, phosphopeptide maps of either the 96 or 93 kDa subunits were identical in intact synaptosomes compared with synaptosomal cytosol. These results show that a phosphoprotein phosphorylated in intact synaptosomes and a 96 000 dalton protein kinase C substrate from rat brain synaptosomal cytosol are the same, and raise the possibility that protein kinase C is the protein kinase responsible for dephosphin phosphorylation in intact synaptosomes.
Cloning of segment polarity gene homologues from the unsegmented brachiopod Terebratulina retusa (Linnaeus)
Authors: Holland Peter W.H. , Williams Nicola A. , Lanfear Jeremy
Published: July 2002

We have used the polymerase chain reaction (PCR) to amplify, clone and sequence homologues of the Drosophila segment polarity genes engrailed (en), cubitus interruptus Dominant (ciD
) and wingless (wg) from the genome of the brachiopod, Terebratulina retusa (Linnaeus). The deduced translation products of brachiopod en and ciD
share high levels of sequence identity with their Drosophila homologues. The brachiopod wg‐related clone is divergent from Drosophila wg, although clearly a member of the wg/Wnt gene family. These results indicate that structural diversity of Drosophila segment polarity genes has been evolutionarily conserved in a divergent, ancient and unsegmented animal phylum.
Site‐directed mutagenesis of a single residue changes the binding properties of the serotonin 5‐HT2 receptor from a human to a rat pharmacology
Authors: Kao Hung-Teh , Adham Nika , Olsen Michael A. , Weinshank Richard L. , Branchek Theresa A. , Hartig Paul R.
Published: July 2002

Mesulergine displays approximately 50‐fold higher affinity for the rat 5‐HT2 receptor than for the human receptor. Comparison of the deduced amino acid sequences of cDNA clones encoding the human and rat 5‐HT2 receptors reveals only 3 amino acid differences in their transmembrane domains. Only one of these differences (Ser → Ala at position 242 of TM5) is near to regions implicated in ligand binding by G protein‐coupled receptors. We investigated the effect of mutating Ser242 of the human 5‐HT2 receptor to an Ala residue as is found in the rat clone. Both [3H]mesulergine binding and mesulergine competition of [3H]ketanserin binding showed high affinity for rat membranes and the mutant human clone but low affinity for the native human clone, in agreement with previous studies of human postmortem tissue. These studies suggest that a single naturally occurring amino acid change between the human and the rat 5‐HT2 receptors makes a major contribution to their pharmacological differences.
The VP35 and VP40 proteins of filoviruses
Authors: Bukreyev A.A. , Volchkov V.E. , Blinov V.M. , Netesov S.V.
Published: July 2002

The fragments of genomic RNA sequences of Marburg (MBG) and Ebola (EBO) viruses are reported. These fragments were found to encode the VP35 and VP40 proteins. The canonic sequences were revealed before and after each open reading frame. It is suggested that these sequences are mRNA extremities and at the same time the regulatory elements for mRNA transcription. Homology between the MBG and EBO proteins was discovered.
Enhancement of phospholipase A2 activation by phosphatidic acid endogenously formed through phospholipase D action in rat peritoneal mast cell
Authors: Sato Takashi , Ishimoto Tsuyoshi , Akiba Satoshi , Fujii Tatsuzo
Published: July 2002

Contribution of phosphatidic acid (PA) generated by activated phospholipase (PL) D to PLA2 activation was studied in rat peritoneal mast cells. Exogenous didecanoyl PA induced arachidonate liberation in the permeabilized cells which was inhibited by p‐bromophenacyl bromide. Upon exposure of the cells to ethanol in a high enough concentration to prevent PA formation, A23187‐induced arachidonate liberation was suppressed by 50% and the rest was completely inhibited by p‐bromophenacyl bromide. In contrast, propranolol, which enhanced PA accumulation, significantly increased the arachidonate liberation. These results suggest that A23187‐induced PLA2 activation may be potentiated, at least in part, by PA generated through PLD action.
Endocytosis and intracellular sorting of ricin and Shiga toxin
Authors: Sandvig Kirsten , van Deurs Bo
Published: July 2002

Ricin and Shiga toxin belong to a group of protein toxins with targets in the cytosol. These toxins consist of one moiety that binds the toxin molecule to cell surface receptors, and another enzymatically active moiety that enters the cytosol after endocytic uptake of the toxin. The toxins are of current interest in relation to disease and the construction of immunotoxins. Moreover, they have proven useful to investigate mechanisms of endocytosis and to follow intracellular pathways of transport. Some of the recent results obtained with ricin and Shiga toxin are discussed.
Crystal structure of macrophage migration inhibitory factor from human Å lymphocyte at 2.1 Å resolution
Authors: Sugimoto Hiroshi , Suzuki Mamoru , Nakagawa Atsushi , Tanaka Isao , Nishihira Jun
Published: July 2002

The three‐dimensional structure of the macrophage migration inhibitory factor (MIF) from human lymphocytes has been determined by X‐ray crystallography at 2.1 A resolution. The structure was solved by a molecular replacement technique using the coordinates of rat MIF. The molecule forms a trimer structure similar to the rat MIF. However, unlike the rat MIF whose C‐terminal tail (residues 104–114) is disordered in the crystal, human MIF has a dermite main‐chain conformation up to the C‐terminal end. These eleven residues create two more β‐strands and join to the inter‐subunit β‐sheet which contribute to forming a trimer structure. Thus, the trimer structure consists of three seven‐stranded β‐sheets surrounded by six α‐helices. Each β‐sheet is comprised of β‐strands from each of the three monomers. This architecture is almost identical to 5‐carboxymethyl‐2‐hydroxymuconate isomerase (CHMI) and is related to the E. coli signal transducing protein PII.
New role of glycosaminoglycans on the plasma membrane proposed by their interaction with phosphatidylcholine
Authors: Satoh Ayano , Toida Toshihiko , Yoshida Keiichi , Kojima Kyoko , Matsumoto Isamu
Published: July 2002

Glycosaminoglycan side chains of membrane proteoglycans have been claimed to be located at the outermost layer of the glycocalyx surrounding the cell. In this study measurements by surface plasmon resonance and solid‐phase assay have shown that both chondroitin sulfate and keratan sulfate but not heparin associate with phosphatidylcholine under physiological conditions. Spectrophotometric measurements also showed that chondroitin sulfate restricts the lateral diffusion of phosphatidylcholine in liposomes. These findings indicate that chondroitin sulfate and/or keratan sulfate chains of membrane proteoglycans crouch on the surface of the membrane while heparan sulfate chains stretch outward from the membrane surface as postulated traditionally.
Expression of T‐ and L‐type calcium channel mRNA in murine sinoatrial node
Authors: Bohn Georg , Moosmang Sven , Conrad Heinke , Ludwig Andreas , Hofmann Franz , Klugbauer Norbert
Published: July 2002

At the cellular level, cardiac pacemaking which sets the rate and rhythm of the heartbeat is produced by the slow diastolic depolarization. Several ion channels contribute to this pacemaker depolarization, including T‐type and L‐type calcium currents. To evaluate the molecular basis of the currents involved, we investigated the cellular distribution of various low voltage activated (LVA) and high voltage activated (HVA) calcium channel mRNAs in the murine sinoatrial (SA) node by in‐situ hybridization. The most prominently expressed LVA calcium channel in the SA node is Cav3.1, whereas Cav3.2 is present at moderate levels. The dominant HVA calcium channel transcript is Cav1.2; only traces of Cav1.3 mRNA are detectable in SA myocytes of mice.
Desialylation of extracellular GD1a‐neoganglioprotein suggests cell surface orientation of the plasma membrane‐bound ganglioside sialidase activity in human neuroblastoma cells
Authors: Kopitz Jürgen , Oehler Christian , Cantz Michael
Published: July 2002

The orientation of the catalytic site of a ganglioside‐specific sialidase in the plasma membrane of SK‐N‐MC neuroblastoma cells was probed using water‐soluble GD1a‐neoganglioprotein substrate on intact cells and GM1‐product detection by cholera toxin B. Desialylation of substrate was readily observed, whereas specific sialidase inhibitors prevented the reaction, and conditioned medium was inactive. Inhibitors of endocytosis and acidification had no effect on substrate degradation, and lowering temperature to 18°C reduced activity but did not abolish it. We conclude that the ganglioside sialidase activity is cell surface‐orientated and displays an in situ specificity that mirrors enzyme preparations in vitro.
Molecular cloning and expression of an adenylyl cyclase from Xenopus laevis oocytes1
Authors: Torrejón Marcela , Echeverrı́a Valentina , Retamales Gabriel , Herrera Luisa , Hinrichs Marı́a V , Olate Juan
Published: July 2002

© 1997 Federation of European Biochemical Societies.
Hedgehog and patched gene expression in adult ocular tissues
Authors: Takabatake Takashi , Ogawa Masanori , Takahashi Tadashi C , Mizuno Makoto , Okamoto Mitsumasa , Takeshima Kazuhito
Published: July 2002

We analysed the expression of members of the hh gene family in adult ocular tissues of newt, frog and mouse by RT‐PCR method. Shh displayed restricted expression in the neural retina that was conserved in each species analyzed. X‐bhh, X‐chh and mouse Ihh were detected in the iris and in the retinal pigment epithelium, while mouse Dhh was detected additionally in the neural retina and faintly in the cornea. We also found that two types of ptc genes, potential hh targets and receptors, were expressed in these tissues, suggesting the presence of active hh signalling there.
Interactions of heterologous DNA with polyomavirus major structural protein, VP1
Authors: Štokrová Jitka , Palková Zdena , Fischer Lukáš , Richterová Zuzana , Korb Jan , Griffin Beverly E. , Forstová Jitka
Published: July 2002

‘Empty' polyomavirus pseudocapsids, self‐assembled from the major structural protein VP1, bind DNA non‐specifically and can deliver it into the nuclei of mammalian cells for expression [Forstová et al. (1995) Hum. Gene Ther. 6, 297–306]. Formation of suitable VP1‐DNA complexes appears to be the limiting step in this route of gene delivery. Here, the character of VP1‐DNA interactions has been studied in detail. Electron microscopy revealed that VP1 pseudocapsids can create in vitro at least two types of interactions with double‐stranded DNA: (i) highly stable complexes, requiring free DNA ends, where the DNA is partially encapsidated; and, (ii) weaker interactions of pseudocapsids with internal parts of the DNA chain.
Uncoupling protein‐3 gene expression in skeletal muscle during development is regulated by nutritional factors that alter circulating non‐esterified fatty acids
Authors: Brun S , Carmona M.C , Mampel T , Viñas O , Giralt M , Iglesias R , Villarroya F
Published: July 2002

Uncoupling protein‐3 gene expression in skeletal muscle is up‐regulated during postnatal development of mice. A high‐carbohydrate diet at weaning induces a decrease in uncoupling protein‐3 mRNA levels that does not occur when mice were weaned onto a high‐fat diet. Uncoupling protein‐3 mRNA levels do not increase in response to fasting in young pups. Only after day 15 of life, when fasting increases serum non‐esterified fatty acids, uncoupling protein‐3 mRNA is up‐regulated by starvation. Over‐nutrition or under‐nutrition during lactation increases or decreases, respectively, uncoupling protein‐3 mRNA expression in skeletal muscle. Regulation of uncoupling protein‐3 gene expression in skeletal muscle during development is mediated by ontogenic and nutritional factors determining changes in circulating non‐esterified fatty acids.
Branched oligonucleotide‐intercalator conjugate forming a parallel stranded structure inhibits HIV‐1 integrase
Authors: Brodin Priscille , Pinskaya Marina , Volkov Evgenii , Romanova Elena , Leh Hervé , Auclair Christian , Mouscadet Jean-François , Gottikh Marina
Published: July 2002

Integration of a DNA copy of the HIV‐1 genome into chromosomal DNA of infected cells is a key step of viral replication. Integration is carried out by integrase, a viral protein which binds to both ends of viral DNA and catalyses reactions of the 3′‐end processing and strand transfer. A 3′‐3′ branched oligonucleotide functionalised by the intercalator oxazolopyridocarbazole at each 5′‐end was found to inhibit integration in vitro. We show that both a specific (G,A) sequence and the OPC intercalating agent contribute to the capability of the branched oligonucleotide to form a parallel stranded structure responsible for the inhibition.
Update on cancer control in women
Authors: Boyle P. , Maisonneuve P. , Autier P.
Published: July 2002

The global cancer burden in women appears to have stabilized according to the most recent estimates available although the distribution of cancer types appears to be changing with a sharp contrast between the increase in the absolute numbers of breast cancers and a decline in cervix cancers. Prospects for cancer control in women appear to be good within our current knowledge and deserve close attention. Rates of lung cancer in women are increasing substantially in many countries and seem set to overtake breast cancer as the commonest form of cancer death in women in many parts of the world. These changes are due to the effects of cigarette smoking, a habit which women widely embraced during the second half of the last century. The high levels of smoking currently in young women, which have yet to have their full impact on death rates, constitute an important hazard not only for future cancer risks but for several other important causes of death. There is strong and consistent evidence that increased consumption levels of fruit and vegetables is associated with reduced risks of many common forms of cancer including breast cancer. Although the breast is the commonest form of cancer in women in most western countries, the etiology of this disease remains elusive and preventable causes remain to be identified. Endogenous hormones also appear to have a role in cancer risk in women: oral contraceptives seem to increase slightly the risk of breast cancer in users in the use and in the immediate post‐use period, but 10 years after cessation the risk again returns to that of never users. Oral contraceptive usage also appears to be protective against ovarian and endometrial cancer. The use of hormonal replacement therapy (HRT) appears to increase the risk of endometrial cancer and a positive association with breast cancer risk appears to exist. Within our current knowledge of the epidemiology of cancer in women, the most important preventive strategies would appear to be the prevention of cigarette smoking and increased dietary intake of vegetables and fruits. Screening has also shown to be effective in reducing incidence and mortality of cervix cancer and mortality from breast cancer. Although more work is needed, it is becoming clear that there could be an important role of HPV testing to further enhance cervix cancer screening. There are important variations in survival from a variety of cancers which are due to factors unrelated to the tumor behavior and that there are significant variations in survival from cancer. Reduction of these gaps could lead to a reduction in cancer mortality and contribute towards increased prospects for cancer control in women.
Clinical management of agitation in the elderly with tiapride
Authors: Robert P.H. , Allain H.
Published: July 2002

Agitated behaviors such as uncooperativeness with necessary care, motor hyperactivity, and verbal or physical aggression are some of the most commonly reported complications in dementia and organic disorders in elderly subjects. These symptoms present greater clinical challenges and management issues than the cognitive deficits. Antipsychotics are the most commonly used psychotropic agents for treating these types of symptoms.

The aims of this article are to review clinical studies with tiapride, a substituted benzamide, and more specifically to present recent data coming from two double-blind, randomized studies in elderly subjects.

The first study versus melperone was conducted in Germany, with over 176 hospitalized demented patients, and indicated that tiapride was as effective and safe as melperone.

More recently, a multicentre, international, double-blind, three-parallel group study compared a 21-day treatment of tiapride to haloperidol and placebo and included 306 demented elderly patients with agitation and aggressiveness. The results showed that tiapride and haloperidol were significantly effective in the treatment of agitation and aggressiveness compared to placebo. The tiapride safety profile was found to be better than haloperidol for clinical acceptability, particularly for significantly fewer extrapyramidal symptoms in the tiapride group.
Mind in Life in Architecture: A Conversation with Architectural Historian Alberto Pérez‐Gómez
Authors: Ritchie Ian
Published: November 2020

In a wide‐ranging interview, Guest‐Editor Ian Ritchie asks Alberto Pérez‐Gómez, the Saidye Rosner Bronfman Professor of the History and Theory of Architecture at McGill University, Montreal, about architecture and human perception. Architecture, Pérez‐Gómez explains, operates as a communicative setting for societies; its beauty and its meaning lie in its connection to human health and self‐understanding.
Resonant Bodies in Immersive Space
Authors: Robinson Sarah
Published: November 2020

Our description and creation of space is nested tightly with our body from which we cannot escape. Our sense of space is dependent on much more than our vision. Architect, writer, editor and lecturer Sarah Robinson explains this particularly in relation to hearing, sound and music. This relationship is symbiotic – we tune our instruments and their harmonics tune us.
Losing Myself: Designing for People with Dementia
Authors: McLaughlin Níall
Published: November 2020

Architect and Professor of Architectural Practice at the Bartlett School of Architecture, University College London, Níall McLaughlin describes his research into dementia and Alzheimer's disease. He explains the thinking behind his practice's design for an Alzheimer's Respite Centre in Dublin, and his subsequent collaboration with fellow Bartlett Professor of Architecture and Experimental Practice Yeoryia Manolopoulou in the creation of an installation about these themes at the Venice Architecture Biennale.
Mind Landscapes: Navigation, Habitat and Imagination
Authors: Corner James
Published: November 2020

Landscapes should be seen not as constituencies of things, animate or inanimate, but as continuous fields, visceral milieux of intersecting social situations, shifting topologies and personal mythologies. The ‘drawing’ of an individual's cognitive map is the residual sediment of their journey through these ambiences. Landscape architect and theorist James Corner explains his conception of landscape and its synthesis with mind.
News from Nowhere: A Biological Reading
Authors: Mallgrave Harry Francis
Published: November 2020

Good architects have always understood that the embodied experience of architecture is multisensory and depends on a variety of parameters that number more than the usually named senses. These parameters are expanding as scientists are beginning to understand the sympathetic resonance between ourselves and others, between our organic bearing and the environment in which we dwell. Harry Frances Mallgrave, architect, scholar, editor and Distinguished Professor Emeritus at the Illinois Institute of Technology, navigates us through this terrain.
Plant Consciousness: Towards an Architecture of Expanded Kinship
Authors: Todd Andrew
Published: November 2020

Neuroscience and phenomenology are valuable tools for architects who wish to increase our engagement with the world as a commonly lived nexus, rather than as a locus of showing‐off and design‐branding. Andrew Todd, architect, argues that our connection with the world can be extended by deeper consideration of plants and their role in architecture.
Wicked Neuroarchitecture: Reciprocity, Shapeshifting Problems and a Case for Embodied Knowledge
Authors: Zisch Fiona
Published: November 2020

Wicked problems are difficult or impossible to solve because of incomplete, changing or even contradictory conditions. Both architecture and neuroscience work on wicked problems as a matter of course. The utilisation of evidence‐based design by some neuroarchitecture researchers implies that architecture is subservient to neuroscience, simply a matter of designing brain impulses. Fiona Zisch, lecturer in architecture and a neuroarchitecture researcher based in London and Innsbruck, suggests otherwise.
Printing Multi‐Material Organic Haptic Actuators
Authors: Zhai Yichen , Wang Zhijian , Kwon Kye‐Si , Cai Shengqiang , Lipomi Darren J. , Ng Tse Nga
Published: November 2020

Haptic actuators generate touch sensations and provide realism and depth in human–machine interactions. A new generation of soft haptic interfaces is desired to produce the distributed signals over large areas that are required to mimic natural touch interactions. One promising approach is to combine the advantages of organic actuator materials and additive printing technologies. This powerful combination can lead to devices that are ergonomic, readily customizable, and economical for researchers to explore potential benefits and create new haptic applications. Here, an overview of emerging organic actuator materials and digital printing technologies for fabricating haptic actuators is provided. In particular, the focus is on the challenges and potential solutions associated with integration of multi‐material actuators, with an eye toward improving the fidelity and robustness of the printing process. Then the progress in achieving compact, lightweight haptic actuators by using an open‐source extrusion printer to integrate different polymers and composites in freeform designs is reported. Two haptic interfaces—a tactile surface and a kinesthetic glove—are demonstrated to show that printing with organic materials is a versatile approach for rapid prototyping of various types of haptic devices.
A Function‐Separated Design of Electrode for Realizing High‐Performance Hybrid Zinc Battery
Authors: Zhong Yijun , Xu Xiaomin , Liu Pengyun , Ran Ran , Jiang San Ping , Wu Hongwei , Shao Zongping
Published: November 2020

A rechargeable hybrid zinc battery is developed for reaching high power density and high energy density simultaneously by introducing an alkaline Zn–transition metal compound (Zn–MX) battery function into a Zn–air battery. However, the conventional single‐layer electrode design cannot satisfy the requirements of both a hydrophilic interface for facilitating ionic transfer to maximize the Zn–MX battery function and a hydrophobic interface for promoting gas diffusion to maximize the Zn–air battery function. Here, a function‐separated design is proposed, which allocates the two battery functions to the two faces of the cathode. The electrode is composed of a hydrophobic MnS layer decorated with Ni–Co–S nanoclusters that allows for smooth gas diffusion and efficient oxygen electrocatalysis and a hydrophilic NixCo1−xS2 layer that favors fast ionic transfer and superior performance for energy storage. The battery with the function‐separated electrode shows a high short‐term discharge voltage of ≈1.7 V, an excellent high‐rate galvanostatic discharge–charge with a power density up to 153 mW cm−2 at 100 mA cm−2, a good round‐trip efficiency of 75% at 5 mA cm−2, and a robust cycling stability for 330 h with an excellent voltage gap of ≈0.7 V at 5 mA cm−2.
Growth decline mechanism of Zanthoxylum planispinum var. dintanensis in the canyon area of Guizhou Karst Plateau
Authors: Yu Yanghua , Song Yanping , Zhong Xinping , Li Yitong , Ying Bin
Published: November 2020

Zanthoxylum planispinum var. dintanensis is an ecological and economic tree species widely planted in the canyon area of the Guizhou Karst Plateau. However, the rapid growth decline of its plantation in large scale has become one of the major environmental problems affecting vegetation restoration and the consolidation of the achievements of rocky desertification control. In this study, we collected soil samples in the full bloom period (March) and the vigorous growth period (June) from the declining and normal‐growing Z. planispinum var. dintanensis communities. Through analyzing fertility quality indexes such as water content and nutrient levels, the mechanism of water–fertilizer coupling affecting the growth decline of Z. planispinum var. dintanensis plantation was explored. The results showed that the pH of soil was about 7.0, and there was no significant difference between the two types of communities. The surface soil water content of the declining community was <20%, which was significantly lower than that of the normal‐growing Z. planispinum var. dintanensis community. The deficiency of quick‐acting nutrients (e.g., available Ca, Mg, and B) and the imbalance of the medium and trace elements could cause the growth decline of Z. planispinum var. dintanensis, indicating that we should pay attention to the addition of quick‐acting nutrients and their stoichiometry in the management process of Z. planispinum var. dintanensis. The comprehensive analysis showed that the decrease of soil water content induced the water–fertilizer coupling deficit effect, which might be one of the reasons for the growth decline of the Z. planispinum var. dintanensis community, in view of the fact that there are many factors causing the growth decline of artificial forest and the mechanism is complex. In the future, it is necessary to systematically study the growth decline of Z. planispinum var. dintanensis community from the perspectives of environmental media, root exudates, plant endogenous hormones, soil microorganisms, and water–fertilizer coupling based on the variations in time and space.
Micro‐computed tomography for the 3D time‐resolved investigation of monodisperse droplet generation in a co‐flow setup
Authors: Schuler Julia , Neuendorf Laura Maria , Petersen Kai , Kockmann Norbert
Published: November 2020

Droplet generation in microfluidic devices has emerged as a promising approach for the design of highly controllable processes in the chemical and pharmaceutical industry. However, droplet generation is still not fully understood due to the complexity of the underlying physics. In this work, micro‐computed tomography is applied to investigate droplet formation in a circular channel in a co‐flow configuration at different flow conditions (Ca < 0.001). The application of an in‐house developed scanning protocol assisted by comprehensive image processing allows for the time‐resolved investigation of droplet formation. By tracking different droplet parameters (length, radii, volume, surface, Laplace pressure) the effect of flow conditions on droplet progression is determined. As characteristic for the squeezing regime, final droplet size was nearly independent of Ca for higher Ca tested. For lower Ca, the final droplet size increased with decreasing Ca, which points to the leaking regime that was recently introduced in the literature.