bluuebunny/crossref_metadata_2025 huggingface dataset:
The relationship between changes in the redox state of plastoquinone and control of excitation energy distribution in photosynthesis
Authors: Dominy Peter , Williams W.Patrick
Published: July 2002

The hypothesis that excitation energy distribution between PS I and PS II is controlled by the redox state of the plastoquinone pool between the two photosystems was investigated using the green algaChlorella vulgaris. Changes in the redox state of the pool were monitored by measurement of the area above the fluorescence induction curve on exposure to high‐intensity light. In agreement with the hypothesis, exposure of state I adapted cells to light preferentially absorbed by PS II led to a reduction of the plastoquinone pool whilst exposure of State II adapted cells to light preferentially absorbed by PS I resulted in its oxidation. However, the limits within which these fluctuations occurred were much narrower than anticipated. The reasons for this are discussed in terms of the possible involvement of changes in the redox state of more specialised molecules associated with the main plastoquinone pool and the postulated role of plastoquinone as an electron shuttle between the two photosystems.
Effect of preillumination on the P‐680+reduction kinetics in chloride‐free photosystem II membranes
Authors: Ono Taka-aki , Conjeaud Helene , Gleiter Hermann , Inoue Yorinao , Mathis Paul
Published: July 2002

The re‐reduction course of P‐680+, the photooxidized PS II primary donor, was measured as a function of excitation number in Cl−‐depleted PS II membranes. After the 1st and 2nd excitations the signal amplitude of P‐680+is small, indicating a submicrosecond reduction of P‐680+by Z, the secondary donor of PS II. After the 3rd excitation, however, a larger P‐680+signal with a 40–50 μs half‐life is observed. The slow decay of this signal is attributed to a back‐reaction with a reduced acceptor in the presence of the Z+S2state on the donor side. The state Z+S2has a lifetime longer than 300 ms and its formation was found to depend on the presence of the abnormal S2state created by the 1st excitation. The P‐680 data and thermoluminescence measurements show that the S‐state advancement beyond S2is blocked in the absence of Cl−and that the Cl−‐free abnormal S2state has a lifetime about 10‐times longer than the normal S2state.
Flash‐induced absorption changes in photosystem I at low temperature: evidence that the electron acceptor A1is vitamin K1
Authors: Brettel K. , Sétif P. , Mathis P.
Published: July 2002

Low temperature flash absorption spectroscopy has been applied to elucidate the chemical nature of the secondary electron acceptor A1of photosystem I (PS‐I). The flash‐induced absorption changes measured in digitonin‐fractionated spinach PS‐I particles at 10 K between 240 and 525 nm are shown to comprise a major decay phase witht~ 150 μs which has been attributed to the recombination reaction P‐700+·A1→ P‐700·A1[(1984) Biochim. Biophys. Acta 767, 404‐414]. We present the absorption difference spectrum of this reaction and demonstrate that it contains contributions in the ultraviolet due to A1, which are characteristic of vitamin K1(phylloquinone).
Complete tracking of proton flow in thylakoids‐the unit conductance of CF0is greater than 10 fS
Authors: Schoenknecht Gerald , Junge Wolfgang , Lill Holger , Engelbrecht Siegfried
Published: July 2002

We investigated the proton conductance of the channel portion of chloroplast ATP synthase (CF0). Thylakoids were CF1‐depleted by EDTA treatment. Proton pumps were stimulated by short flashes of light. Proton flux through CF0was measured spectrophotometrically in three different ways: as proton efflux from the lumen (via neutral red), charge flow across the membrane (via electrochromism) and proton influx into the medium (via phenol red). Hence we completely tracked the protons on their way from the lumen through CF0into the medium. A first treatment with EDTA removed up to 12% of total CF1without increasing the proton permeability of the membranes. A second treatment removed a further 20% of CF1and increased the proton permeability of membranes by 3 orders of magnitude. The electric potential difference and the pH transients, in both the lumen and medium, decayed with a relaxation time of 7 ms indicating electrically driven proton flow through CF0. If the electric driving force was shunted (e.g. by added gramicidin) both pH transients decayed at 85 ms compared with 20–60 s in control thylakoids. The longer relaxation time under chemical driving force was attributable to larger chemical than electrical capacitance of thylakoids. We calculated a lower limit of the unit conductance of CF0under the assumption that all exposed CF0were proton conducting. The value was 10 fS, corresponding to the passage of 6200 protons/s per CF0(at 100 mV electric driving force) and by orders of magnitude higher than so far reported for any F0channel.
On the function of two vitamin K1molecules in the PS I electron acceptor complex
Authors: Malkin Richard
Published: July 2002

Various PS I preparations from higher plants and cyanobacteria are found to contain vitamin K1, or phylloquinone, in a stoichiometry of 2 vitamin K1/P700. Extractions of lyophilized PS I preparations with organic solvents led to removal of one molecule of vitamin K1but conditions have not been established for the removal of the second quinone. The quinone‐depleted PS I complex ( ~ 1 vitamin K1/P700) was fully active in transferring electrons at both physiological and cryogenic temperatures, indicating one vitamin K1molecule was not required for these activities.
The influence of copper on the induction of tyrosinase and laccase in Neurospora crassa
Authors: Huber Marcel , Lerch Konrad
Published: July 2002

The influence of copper on the cycloheximide‐induced synthesis of the copper‐containing enzymes tyrosinase and laccase in Neurospora crassa was studied by enzyme activity measurements and immunological means. The amount of active enzyme molecules is far higher when the culture medium is copper‐supplemented before cycloheximide induction. The synthesis of the apoproteins is not dependent on the presence of copper. This suggests the existence of a copper‐storage protein for which metallothionein is a likely candidate.
Primary structures of human protein kinase C βI and βII differ only in their C‐terminal sequences
Authors: Kubo Kyoko , Ohno Shigeo , Suzuki Koichi
Published: July 2002

Two types of cDNA clones encoding human protein kinase C (PKC) were isolated from a spleen cDNA library using rabbit protein kinase C βI/βII cDNA as a hybridization probe. Nucleotide sequence analyses of these cDNA inserts revealed complete primary structures of two distinct types of human protein kinase C βI and βII which differ only in their C‐terminal 50 or 52 amino acid residues. It was concluded that there exist four distinct types of PKC, PKC α, βI, βII and γ, in human as well as rabbit, and that the corresponding sequences are strictly conserved among mammalian species.
Bound electron acceptors of photosystem I Evidence against the identity of redox center A1with phylloquinone
Authors: Ziegler K. , Lockau W. , Nitschke W.
Published: July 2002

Photosystem (PS) I preparations from spinach and from a cyanobacterium contain two molecules of phylloquinone (2‐methyl‐3‐phytyl‐1,4‐naphthoquinone) per photosystem, only one of which is rapidly destroyed by ultraviolet light. In preparations of the core of PS I, the P700 reaction center, both quinone molecules are rapidly destroyed by UV irradiation. Nearly complete destruction does not alter the shape and size of the EPR spectrum of the electron acceptor A1, which is believed to be phylloquinone. Photooxidation of P700, the primary electron donor of PS I, is also not affected. The results provide strong evidence against the identity of redox center A1with the naphthoquinone.
EPR study of charge equilibrium at low temperatures in the S2state of oxygen‐evolving photosystem II particles
Authors: Kawamori Asako , Satoh Jun-ichi , Inui Takashi , Satoh Kimiyuki
Published: July 2002

The stability of EPR Signal IIsin oxygen‐evolving PS II particles in the different S‐states has been studied as a function of storage time at 77 K. Signal IIsin dark‐adapted samples retains the same intensity after 1 month storage at 77 K, while that in the S2state produced by illumination at 195 K decays with a half‐life time of about 5 days without decay in the manganese multiline intensity observed at 4.5 K. Signal IIs, once decayed in the S2state, recovers its original intensity upon dark adaptation of the sample at 210 K accompanied by a decrease in the multiline intensity. A model for the charge recombination process for the S2state involving Signal IIsis discussed.
The expression in E. coli of a polymeric gene coding for an esterase mimic catalyzing the hydrolysis of p‐nitrophenyl esters
Authors: Bülow Leif , Mosbach Klaus
Published: July 2002

We have prepared a hybrid protein consisting of seven esterase units, Glu‐Ala‐His‐Ala‐Ser‐Phe‐Phe‐Phe, fused to the N‐terminal of galactokinase (E. coli). The structural gene for this bifunctional protein was obtained by cloning a polymer made up of three chemically synthesized oligonucleotides to which the galactokinase gene was fused in frame. The hybrid protein was purified to homogeneity with the aid of the galactokinase moiety and showed an M
r of 51 000‐53 000. The preparation could catalyze the hydrolysis of p‐nitrophenyl esters and, due to the inbuilt hydrophobic spacers, Phe‐Phe‐Phe, improved catalysis of more hydrophobic substrates was obtained.
Transbilayer organization of the main chlorophylla/b‐protein of photosystem II of thylakoid membranes
Authors: Anderson Jan.M. , Goodchild D.J.
Published: July 2002

To probe the location of the carboxyl‐terminus of the 28 kDa apoprotein of the light‐harvesting chlorophylla/b‐protein complex of PS II (LHCII), an antibody was generated against a synthetic octapeptide corresponding to the C‐terminal region of LHCII. The high specificity of the octapeptide antiserum was deonstrated by immunoblots and immunogold labelling. The octapeptide antiserum agglutinated destacked thylakoid membranes, but no significant agglutination occurred with inside‐out vesicles suggesting that the COOH‐terminus is located at the outer, stroma‐exposed surface where the NH2‐terminus is also located [(1983) J. Biol. Chem. 258, 9941‐9948]. Our results support a model for LHCII with four transmembrane‐spanning domains.
Clostridium botulinum type C produces a novel ADP‐ribosyltransferase distinct from botulinum C2 toxin
Authors: Aktories Klaus , Weller Ulrich , Chhatwal Gursharan S.
Published: July 2002

The culture medium of certain strains of Clostridium botulinum type C contains two separable ADP‐ribosyltransferases. Besides the ADP‐ribosylation of actin due to botulinum C2 I toxin, a second microbial enzyme causes the mono‐ADP‐ribosylation of a eukaryotic protein with a molecular mass of about 20 kDa found in platelets, neuroblastoma × glioma hybrid cells, S49 lymphoma cells, chick embryo fibroblasts and sperm. The eukaryotic substrate is inactivated by heating and trypsin treatment. In contrast, the novel ADP‐ribosyltransferase, which can be separated by DEAE‐Sephadex chromatography, is largely resistant in the short term to trypsin digestion.
M13 phage DNA as a universal marker for DNA fingerprinting of animals, plants and microorganisms
Authors: Ryskov A.P. , Jincharadze A.G. , Prosnyak M.I. , Ivanov P.L. , Limborska S.A.
Published: July 2002

Hypervariable polymorphic patterns were detected with M13 phage DNA as a probe in genomic DNA of organisms belonging to different taxonomic groups including animals (vertebrates and invertebrates), plants and microorganisms. Individual‐specific restriction pattern analysis (DNA fingerprinting) with this probe proved to be useful for individual identification, analysis of somatic stability and paternity testing in man. The nuclear type of inheritance indicates that the hypervariable DNA regions in question are located in the chromosomes, not in the mitochondrial DNA. The data obtained also demonstrate a potential range of M13 DNA applications as a probe for DNA fingerprinting of animals, plants and microorganisms, particularly for the determination of inbred lines, identification of bacterial strains and establishing stock, variety and strain distinctions.
Do mitochondrial DNA fragments promote cancer and aging?
Authors: Richter Christoph
Published: July 2002

Reactive oxygen species are important in carcinogenesis, diseases, and aging, probably through oxidative damage of DNA. Our understanding of this relationship at the molecular level is very sketchy. It has recently been found that in mitochondria oxidative DNA damage is particularly high and may not be repaired efficiently. I propose that oxidatively generated DNA fragments escape from mitochondria and become integrated into the nuclear genome. This may transform cells to a cancerous state. Time‐dependent nuclear accumulation of mitochondrial DNA fragments may progressively change the nuclear information content and thereby cause aging. This proposal can be tested experimentally.
New series of lipoxins isolated from human eosinophils
Authors: Steinhilber Dieter , Roth Hermann J.
Published: July 2002

Granulocytes from human eosinophilic donors were incubated with arachidonic acid or 15‐hydroxyeicosatetraenoic acid (15‐HETE) and stimulated with the ionophore A23187. The eicosanoids were extracted with reversed‐phase cartridges and subjected to RP‐HPLC analysis. When extracts from eosinophil‐enriched populations were analysed and compared with extracts from human neutrophils, three additional peaks were detected which coeluted with 15‐hydroxy‐Δ13‐trans‐15H derivatives of leukotriene C4, D4 and E4 in different HPLC systems. The recorded absorbance spectra of the eluted compounds and the standards were identical and showed a maximum at 307 nm which is characteristic for a conjugated tetraene system with a bathochromic shift by the sulfur moiety in α‐position to the tetraene system. The compound which coeluted with the 15‐hydroxy‐LTC4 standard was treated with γ‐glutamyltransferase and converted to the corresponding leukotriene D4 derivative. The results indicate that interaction between the 5‐ and 15‐lipoxygenase pathways leads to the formation of a new series of arachidonic acid metabolites in human eosinophils. Since the biosynthetic route is similar to that of lipoxin A4 and lipoxin B4, we suggest the trivial names lipoxin C4, D4 and E4.
Calcium‐dependent DNA fragmentation in human synovial cells exposed to cold shock
Authors: Perotti Mariarosa , Toddei Filippo , Mirabelli Francesca , Vairetti Mariapia , Bellomo Giorgo , McConkey David J. , Orrenius Sten
Published: July 2002

Exposure of confluent human synovial McCoy's cells to near‐freezing temperatures followed by rewarming at 37°C resulted in endonuclease activation and cell death characteristic of a suicide process known as apoptosis. Both DNA fragmentation and cell killing were dependent on a sustained increase in the cytosolic Ca2+ concentration. Sensitivity to cold shock‐induced endonuclease activation was critically dependent on the cell cycle (proliferative) status and limited to confluent cells, whereas cells in the logarithmic growth phase were completely resistant. However, DNA fragmentation was promoted in the proliferating McCoy's cells pretreated with H‐7 or sphingosine, inhibitors of protein kinase C. In addition, phorbol ester, known to activate PKC, inhibited DNA fragmentation in the confluent cells. Our findings indicate that cold shock‐induced DNA fragmentation in McCoy's cells is dependent on a sustained Ca2+ increase, and sensitivity to the process appears to be regulated by the status of protein kinase C.
Human recombinant interleukin‐1 receptor antagonist inhibits lymphocyte blastogenesis induced by concanavalin A Restorative effect of hrIL‐1
Authors: Conti Pio , Reale Marcella , Panara Maria R. , Barbacane Renato C. , Bongrazio Mauro , Dempsey Roy A.
Published: July 2002

Interleukin‐1 (IL‐1), mainly produced by monocyte‐macrophages, is a polypeptide cytokine with pleiotropic biological effects. IL‐1 plays an important role in mediating immune response and inflammation. Recently a natural inhibitor to IL‐1 has been discovered, interleukin‐1 receptor antagonist (IL‐1ra), produced by human monocytes cultured on adherent IgG which binds to the IL‐1 receptors. In our study we found that the pretreatment of cells with serial dilutions of IL‐1ra (250 ng/ml−2.5 pg/ml) inhibits, in a dose‐dependent manner, lymphocyte DNA synthesis stimulated with Con A (10 μg/ml). IL‐1ra did not have any effect on resting peripheral blood mononuclear cells (PBMC). Time course experiments show that IL‐1ra at 250 ng/ml has its maximum inhibitory effect on lymphocyte blastogenesis when cells are pretreated 2 h before Con A. No effect was found when hrIL‐1ra was added after Con A. Moreover, hrIL‐1ra also inhibits the enhancing effects of exogenous hrIL‐1 (400, 200, 100 and 50 ng/ml) on lymphocytes stimulated with Con A; while when hrIL‐1ra was used on cells treated with only Con A, the inhibition was more pronounced. When PBMC were removed from monocytes, by adherence, the Con A‐treated lymphocytes were not influenced by 2 h pretreatment of hrIL‐1ra; while a strong inhibition was found when exogenous hrIL‐1 was added at different concentrations. In addition, hrIL‐1ra also inhibits the enhancing effect of hrIL‐2 on lymphocyte DNA synthesis. In another set of experiments PBMC were pretreated with hrIL‐1ra (250 ng/ml) for 2 h and then added LPs (10 ng/ml) and IL‐1α generation was determined using ELISA. In these experiments IL‐1ra completely abolished the generation of IL‐1α. These data suggest that hrIL‐1ra exhibits a dose—response inhibition of lymphocyte blastogenesis induced by Con A, probably through the down‐regulation of IL‐1 synthesis necessary as an early signal for T‐cell activation and IL‐2 production.
An endothelin B receptor‐selective antagonist: IRL 1038, [Cys11‐Cys15]‐endothelin‐1(11–21)
Authors: Urade Yoshihiro , Fujitani Yasushi , Oda Kyoko , Watakabe Tadashi , Umemura Ichiro , Takai Michihiro , Okada Toshikazu , Sakata Kiyoshi , Kiraki Hideaki
Published: July 2002

In the inhibition of specific binding or [125]endothelins (ETs) to membranes from various tissues of rats, guinea pigs, pigs and humans, [Cys11‐Cys15]‐ET‐1(11–21), IRL 1038, has a much higher affinity for ETB receptors (K
i = 6–11 nM) than for ETA receptors (K
i = 0.4–0.7 μM). In contraction assays, with ET‐3 as a stimulant, 3 μM IRL 1038 antagonized the ETB receptor‐mediated contraction of guinea pig ileal and tracheal smooth muscle without any significant agonistic activity, but did not effect the ETA receptor‐mediated contraction of rat aortic smooth muscle. IRL 1038 is, therefore, considered to be the first antagonist selective to the ETB receptor.
Purification and characterization of a 90 kDa protein released from human tumors and tumor cell lines
Authors: Iacobelli S. , Bucci I. , D'Egidio M. , Giuliani C. , Natoli C. , Tinari N. , Rubistein M. , Schlessinger J.
Published: July 2002

A novel tumor‐associated protein, termed 90K, and recognized by mAb SP‐2 was purified from serum of breast cancer patients, ovarian cancer ascitic fluid and conditioned medium of human breast cancer cells. In these three sources, native 90K is present as a high molecular weight complex that was dissociated by SDS‐PAGE into a major band of approximately 90,000 Da. On the basis of electrophoretic mobility, buoyant density value, amino acid composition, and immunoreactivity, the 90K from the different sources appeared to be identical. NH2‐tenninal amino acid sequence revealed no homology to known protein.
Mechanism of the cytolytic action of Pseudomonas aeruginosa cytotoxin: Oligomerization of the cytotoxin on target membranes
Authors: Ohnishi Makoto , Hayashi Tetsuya , Tomita Toshio , Terawaki Yoshiro
Published: July 2002

Pseudomonas aeruginosa cytotoxin (CTX) is thought to be a pore‐forming polypeptide of 29 kDa. To study whether CTX assembles into oligomer on target membranes, we solubilized membrane‐bound toxin with 1% sodium dodecyl sulfate (SDS) at 25°C and analyzed its molecular size using SDS‐polyacrylamide gel electrophoresis and immunoblot analysis. The results indicate that CTX forms a complex of approximately 145 kDa on the surface of erythrocytes and lipid vesicles, and that the complex formation is closely correlated with the toxin‐induced permeabilization of target membranes. Thus, CTX may assemble into a pore‐forming oligomer on target membranes.
Lactosylceramide is synthesized in the lumen of the Golgi apparatus
Authors: Lannert Heinrich , Bünning Garsten , Jeckel Dieter , Wieland Felix T.
Published: July 2002

Recently, synthesis of lactosylceramide has been described to occur on the cytosolic face of the Golgi [(1991) J. Biol. Chem. 266, 20907‐20912]. The reactions following in the biosynthesis of higher glycosphingolipids are known to take place in the lumen of the Golgi. For our understanding of the functional organization of the multiglycosyltransferase system of glycosphingolipid synthesis in the Golgi, the knowledge of the topology of individual reactions is a prerequisite. We have developed a simple and quick assay system for sphingolipid biosynthesis and have obtained evidence that lactosylceramide is synthesized in the lumen of the Golgi. Because lactosylceramide is generated by galactosylation of glucosylceramide which, in turn, is synthesized from ceramide and UDP‐Glc on the cytosolic surface of the Golgi apparatus, further efforts will be directed to the characterization of a glucosylceramide‐translocator in the Golgi membranes rather than a lactosylceramide‐translocator.
Structural and enzymological analysis of the interaction of isolated domains of cytochrome P‐450 BM3
Authors: Munro Andrew W. , Lindsay J.Gordon , Coggins John R. , Kelly Sharon M. , Price Nicholas C.
Published: July 2002

The interactions of the individually expressed haem‐ and flavin‐containing domains of cytochrome P‐450 BM3 have been analysed by enzymological and spectroscopic techniques. Electron transfer between the isolated domains occurs at a much lower rate than that occurring in the intact flavocytochrome. CD spectroscopic studies indicate that the linkage of the domains in intact P‐450 BM3 creates haem and amino acid environments suitable for efficient electron transfer from its flavin domain.
Glutathione depletion induces glycogenolysis dependent ascorbate synthesis in isolated murine hepatocytes
Authors: Braun László , Csala Miklós , Poussu Anssi , Garzó Tamás , Mandl József , Bánhegyi Gábor
Published: July 2002

The relationship between glutathione deficiency, glycogen metabolism and ascorbate synthesis was investigated in isolated murine hepatocytes. Glutathione deficiency caused by various agents increased ascorbate synthesis with a stimulation of glycogen breakdown. Increased ascorbate synthesis from UDP‐glucose or gulonolactone could not be further affected by glutathione depletion. Fructose prevented the stimulated glycogenolysis and ascorbate synthesis caused by glutathione consumption. Reduction of oxidised glutathione by dithiothreitol decreased the elevated glycogenolysis and ascorbate synthesis in diamide or menadione treated hepatocytes. Our results suggest that a change in GSH/GSSG ratio seems to be a sufficient precondition of altering glycogenolysis and a consequent ascorbate synthesis.
Modulation of the α1A Ca2+ channel by β subunits at physiological Ca2+ concentration
Authors: Cens Thierry , Mangoni Matteo E. , Nargeot Joel , Charnet Pierre
Published: July 2002

The class A Ca2+ channel αl subunit (αlA) was expressed in Xenopus oocytes alone or in combination with the βlb, β2a, β3, or β4 subunit. Analysis of voltage‐dependent activation and inactivation in the presence of 1.8 mM external Ca2+ showed an hyperpolarising shift of both relations when compared to similar recordings performed in the presence of 40 mM Ba2+. These shifts, which differed for activation and inactivation, were strongly modulated by the nature of the co‐expressed β subunit. On the other hand, for each combination, the kinetics of inactivation were similar in 1.8 mM Ca2+ and 40 mM Ba2+ (for example co‐expression of the β2a subunit reduced inactivation using either 40 mM Ba2+ or 1.8 mM Ca2+). Thus, modulation of channel properties by the β subunit is different in physiological Ca2+ or high Ba2+ concentrations. These results must be taken into consideration to extrapolate the role of the β subunit in native cells.
Feasibility of pharmacotherapy in HIV-related affective spectrum disorders: an open trial with fluvoxamine
Authors: Perretta P , Nisita C , Zaccagnini E , Lorenzetti C , Nuccorini A , Cassano GB , McNair DM , Akiskal HS
Published: July 2002

Among HIV patients treated for AIDS-related adjustment, major depressive disorders and other affective disorders, we assessed in an open study the feasibility of using a serotonergic antidepressant (fluvoxamine). Thirty-five seropositive patients with the above conditions (22 men and 13 women) were followed over a minimum period of four weeks. At the end of the treatment, a large number of patients (77%) showed marked improvement. “Nuclear” depressive and anxiety symptoms remitted, while the “somatic” ones seemed less sensitive to treatment. Treatment had to be terminated prematurely due to side effects in only two patients (6%).
An NMR‐based metabonomic approach to the investigation of coelomic fluid biochemistry in earthworms under toxic stress
Authors: Bundy J.G , Osborn D , Weeks J.M , Lindon J.C , Nicholson J.K
Published: July 2002

The endogenous metabolites of the coelomic fluid of the earthwormEisenia venetawere characterised using high‐resolution one‐dimensional and two‐dimensional1H nuclear magnetic resonance spectroscopy. Signals from common organic acids, such as acetate, fumarate, malonate, malate, formate, and succinate, were identified together with adenosine and nicotinamide mononucleotide. The potential use of this information as a baseline dataset for future toxicological or physiological studies was demonstrated by a metabonomic analysis: a series of earthworms were dosed with the model compound 3‐fluoro‐4‐nitrophenol, and toxic effects followed by multivariate analysis of the spectral data of the coelomic fluid. Relative concentrations of acetate and malonate were decreased in the dosed worms compared to the controls.
The 5′ untranslated region of cucumber mosaic virus RNA4 confers highly competitive activity on heterologous luciferase mRNA in cell‐free translation
Authors: Kwon Chang Seob , Paek Kyung Hee , Chung Won-Il
Published: July 2002

The cell‐free translation of virion RNAs of several tripartite RNA viruses has shown that RNA4, a subgenomic RNA, is more competitive than other virion RNAs. Recently, the 3′ untranslated region (UTR) of alfalfa mosaic virus (AMV) RNA4 was identified to be a competitive determinant. In this study, we observed that the RNA4 of cucumber mosaic virus (CMV), another tripartite RNA virus, was also found to be a strong competitor in translational competition among CMV virion RNAs. To identify the competitive determinant of CMV RNA4, we constructed various chimeric luciferase mRNAs containing RNA4 and/or vector‐derived UTRs. The relative translations of luciferase‐containing mRNA in the presence of a competitor mRNA showed that the 5′ UTR, not the 3′ UTR, substantially contributed to the highly competitive activity of CMV RNA4.
Large quantity production with extreme convenience of human SDF‐1α and SDF‐1β by a Sendai virus vector
Authors: Moriya Chikaya , Shioda Tatsuo , Tashiro Kei , Nagasawa Takashi , Ikegawa Masaya , Ohnishi Yukano , Kato Atsushi , Hu Huiling , Xin Xiaomi , Hasan Mohammad K , Maekawa Midori , Takebe Yutaka , Sakai Yuko , Honjo Tasuku , Nagai Yoshiyuki
Published: July 2002

We describe a robust expression of human stromal cell‐derived factor‐1α (SDF‐1α) and SDF‐1β, the members of CXC‐chemokine family, with a novel vector system based upon Sendai virus, a non‐segmented negative strand RNA virus. Recombinant SDF‐1α and SDF‐1β were detected as a major protein species in culture supernatants, reached as high as 10 μg/ml. This remarkable enrichment of the products allowed us to use even the crude supernatants as the source for biological and antiviral assays without further concentration nor purification and will thus greatly facilitate to screen their genetically engineered derivatives.
A novel importin α from rice, a component involved in the process of nuclear protein transport1
Authors: Iwasaki Toshisuke , Matsuki Rikyu , Shoji Kazuhiro , Sanmiya Kazutsuka , Miyao Mitsue , Yamamoto Naoki
Published: July 2002

In eukaryotes, nuclear proteins that are transported into nuclei have nuclear localization signals (NLSs), which are recognized by proteins called importin α. We isolated a rice cDNA, #61L, and the corresponding gene that encodes a protein, which shows significant homology to the importin α. Although the encoded protein had only 23–27% amino acid identity to the importin αs from various organisms including plants, the fusion protein with glutathione S‐transferase showed a specific binding activity to the NLS of SV40 T‐antigen. These results suggest that the rice #61L protein is a novel importin α in plants.
Anchoring antibodies to membranes using a diphtheria toxin T domain‐ZZ fusion protein as a pH sensitive membrane anchor
Authors: Nizard Philippe , Liger Dominique , Gaillard Carole , Gillet Daniel
Published: July 2002

We have constructed a fusion protein, T‐ZZ, in which the IgG‐Fc binding protein ZZ was fused to the C‐terminus of the diphtheria toxin transmembrane domain (T domain). While soluble at neutral pH, T‐ZZ retained the capacity of the T domain to bind to phospholipid membranes at acidic pH. Once anchored to the membrane, the ZZ part of the protein was capable of binding mouse monoclonal or rabbit polyclonal IgG. Our results show that the T‐ZZ protein can function as a pH sensitive membrane anchor for the linkage of IgG to the membrane of lipid vesicles, adherent and non‐adherent cells.
Activation of haptoglobin gene expression by cAMP involves CCAAT/enhancer‐binding protein isoforms in intestinal epithelial cells
Authors: Pelletier Nadine , Boudreau François , Yu Shun-Jiang , Zannoni Sonia , Boulanger Véronique , Asselin Claude
Published: July 2002

CCAAT/enhancer‐binding protein (C/EBP) isoforms are expressed in rodent intestine and in the rat intestinal epithelial cell line IEC‐6 but their role remains to be determined. Treatment of IEC‐6 cells with the adenylate cyclase activator forskolin led to coordinate induction of C/EBP isoforms α, β and δ at the mRNA and protein levels. Transient transfection assays showed that their expression is controlled at the transcriptional level. Forskolin treatment induced haptoglobin mRNA levels. Electrophoretic mobility shift and supershift assays demonstrated an increase in DNA‐binding activities of the three C/EBP isoforms to the haptoA and haptoC C/EBP DNA‐binding sites of the proximal haptoglobin promoter. Site‐specific mutations of both sites led to a decrease in transcriptional induction by forskolin, suggesting that C/EBP isoforms are involved in the cAMP‐dependent regulation of the acute‐phase protein gene haptoglobin in intestinal epithelial cells.
Tumor‐derived EMMPRIN (extracellular matrix metalloproteinase inducer) stimulates collagenase transcription through MAPK p38
Authors: Lim Melissa , Martinez Tom , Jablons David , Cameron Robert , Guo Huiming , Toole Bryan , Li Jian-dong , Basbaum Carol
Published: July 2002

EMMPRIN (extracellular matrix metalloproteinase inducer) stimulates fibroblast metalloproteinases (MMP) 1, 2 and 3 (Kataoka et al. (1993) Cancer Res. 53, 3154–3158). Here we focus on MMP‐1, showing that in lung tumors, MMP‐1's cognate mRNA is strongly expressed in stromal fibroblasts adjacent to EMMPRIN‐expressing tumor cells. In vitro, EMMPRIN upregulates MMP‐1 mRNA expression in a concentration‐dependent manner, with a peak accumulation at 24 h. The response is genistein‐sensitive, suggesting it is dependent on tyrosine kinase activity. Analysis of tyrosine phosphorylation‐dependent MAP kinases ERK 1/2, SAPK/JNK, and p38 showed that the activity of p38 but not that of the other 2 kinases was elevated in response to EMMPRIN. That p38 activity was required for EMMPRIN stimulation of MMP‐1 was evident from results showing that the p38 inhibitor SB203580 blocked this response. This is the first available information regarding the mechanism by which tumor‐associated molecules upregulate MMP synthesis in stromal fibroblasts.
Colocalization of phospholipase D1 and GTP‐binding‐defective mutant of ADP‐ribosylation factor 6 to endosomes and lysosomes
Authors: Toda Kyoko , Nogami Masahiro , Murakami Kazuo , Kanaho Yasunori , Nakayama Kazuhisa
Published: July 2002

Phospholipase D (PLD) is involved in various aspects of cellular function. Two isoforms, PLD1 and PLD2, have been identified. PLD1, which has two splicing variants, is regulated by various factors, including ADP‐ribosylation factor (ARF). We here show that both variants of PLD1 are predominantly localized to late endosomes and lysosomes, but not to the Golgi apparatus or endoplasmic reticulum in contrast to earlier studies. Furthermore, PLD1s show significant colocalization with an ARF6 mutant defective in GTP binding. The data suggest that PLD1, under the regulation of ARF6, plays a role in the function of endosomes and lysosomes.
P2Z purinoreceptor ligation induces activation of caspases with distinct roles in apoptotic and necrotic alterations of cell death
Authors: Ferrari Davide , Los Marek , Bauer Manuel K.A. , Vandenabeele Peter , Wesselborg Sebastian , Schulze-Osthoff Klaus
Published: July 2002

Myeloic cells express a peculiar surface receptor for extracellular ATP, called the P2Z/P2X7 purinoreceptor, which is involved in cell death signalling. Here, we investigated the role of caspases, a family of proteases implicated in apoptosis and the cytokine secretion. We observed that extracellular ATP induced the activation of multiple caspases including caspase‐1, ‐3 and ‐8, and subsequent cleavage of the caspase substrates PARP and lamin B. Using caspase inhibitors, it was found that caspases were specifically involved in ATP‐induced apoptotic damage such as chromatin condensation and DNA fragmentation. In contrast, inhibition of caspases only marginally affected necrotic alterations and cell death proceeded normally whether or not nuclear damage was blocked. Our results therefore suggest that the activation of caspases by the P2Z receptor is required for apoptotic but not necrotic alterations of ATP‐induced cell death.
Import of precursor proteins into mitochondria from soybean tissues during development
Authors: Murcha Monika W , Huang Tiffany , Whelan James
Published: July 2002

Characterisation of the amount of protein import of the alternative oxidase (AOX) and the FAd precursor proteins (previously shown to use different import pathways) into mitochondria from developing soybean tissues indicated that they displayed different patterns. Import of the AOX declined in both cotyledon and root mitochondria with increasing age, whereas the import of the FAd into cotyledon mitochondria remained high throughout the same period. Using primary leaf mitochondria, it was evident that import of AOX remained high while it declined in cotyledon and root mitochondria. The amount of import of the AOX into mitochondria from different tissues closely matched the amount of the Tom 20 receptor.
Phosphatidyl serine exposure during apoptosis precedes release of cytochrome c and decrease in mitochondrial transmembrane potential
Authors: Denecker Geertrui , Dooms Hans , Van Loo Geert , Vercammen Dominique , Grooten Johan , Fiers Walter , Declercq Wim , Vandenabeele Peter
Published: July 2002

Time kinetics of phosphatidyl serine (PS) exposure were compared to other apoptotic parameters following different apoptotic stimuli. Our data indicate that anti‐Fas treatment of L929sAhFas cells results in rapid exposure of PS, which precedes decrease in mitochondrial transmembrane potential (ΔΨ
m) and release of cytochrome c, indicating that PS exposure occurs independently of these mitochondrial events. Also during TNF‐, etoposide‐ or staurosporine‐mediated apoptosis in PC60 RI/RII cells, PS‐positive cells were observed before they had a decreased ΔΨ
m. However, during growth factor depletion‐induced death of 32D cells, both phenomena seemed to occur at the same time.
Cephalocentesis with the modified Smellie's perforator
Authors: Chanrachakul B , Chittachareon A , Herabutya Y
Published: July 2002

A 37‐year‐old pregnant woman, gravida 2, para 0, was referred to Ramathibodi Hospital at 31 weeks of gestation with the diagnosis of hydrocephalus and polyhydramnios. Repeated ultrasound scans revealed hydrocephalus with macrocephaly (BPD=10.3 cm), polyhydramnios (AFI=31.5), and a suspected esophageal atresia. After counseling, both parents decided not to pursue the pregnancy and requested vaginal delivery. They decided against transabdominal, ultrasonic‐guided cephalocentesis because of its invasiveness, patient's awareness, and pain. Skull decompression with the modified Smellie's perforator was performed after five 400‐μg doses of misoprostol were applied to dilate the cervix. The post‐partum condition was uneventful. Fetal skull decompression with the modified Smellie's perforator and misoprostol for cervix dilation is a useful, simple, and safe procedure that can be performed with no previous experience.
Effects of labor time on secretion time and quantity of breast milk
Authors: Doganay M , Avsar F
Published: July 2002

Objectives: The aim of the study is to investigate the effects of labor time on the secretion time and quantity of breast milk in patients who underwent cesarean sections, and to compare findings of patients with and without labor. Methods: A total of 180 patients, divided into three groups of 60 subjects, who had been hospitalized in Zekai Tahir Burak Maternity Training and Research Hospital between December 1999 and September 2000, were included in this study. Group 1 included patients who developed indications for cesarean during labor, group 2 included patients with previous cesareans who were admitted into the Hospital in active labor status, and group 3 comprised patients with previous cesareans who were dated for section (elective). Statistical analyses were performed using the Mann–Whitney U‐test in Statistical Program for Scientific Studies (spss) for Windows 9.0. Results: In group 1, the lactation start time was 71.2 min (10–180 min) and the amount of milk was 26.2 ml (20–35 ml) while the same parameters were, respectively, 64.2 min (10–180 min) and 31.2 ml (20–40 ml) in group 2 and 136.2 min (45–240) and 26.3 ml (20–35 ml) in the group 3. The results were found to be statistically significant in group 2, whereas they were not significant in groups 1 and 3. Conclusions: The first secretion time of lactation was found to be earlier and the amount of breast milk collected within 24 h was higher in patients who were admitted in active labor (group 2), both parameters being statistically significant.
Ultrasound in gynecology: endometrium
Authors: Bourne T. , Hamberger L. , Hahlin M. , Granberg S.
Published: July 2002

With the development and clinical application of transvaginal transducers/probes (TVS) the sonographic imaging of the endometrium was greatly enhanced compared with abdominal ultrasound. Also, the discomfort of a full bladder associated with abdominal ultrasound could be avoided. A shorter distance between probe and target allowed the use of higher frequency transducers, thereby achieving improved imaging. This review will only discuss the use of TVS, but it does not mean that abdominal ultrasound should not or cannot be used when dealing with the postmenopausal uterus. Transvaginal sonography (TVS) provides a valuable tool for the diagnosis of a wide range of gynecological disorders including those of the uterus and endometrium. The ability of TVS to depict the thickness and morphology of the endometrium has been established in both office and hospital settings. This article will discuss and illustrate the clinical and research applications of transvaginal sonography in relation to the endometrium in both symptomatic and asymptomatic postmenopausal women. The article is of particular relevance today given the number of women who are undergoing transvaginal ultrasonography in the absence of symptoms as a part of their routine check ups. There is a paucity of data relating to the management of apparent ultrasound abnormalities in such women. In symptomatic women or for women at risk of developing endometrial pathology, a technique that could reduce the number of biopsy procedures would be of value. Hysteroscopy, dilatation and curettage (D & C) as well as other endometrial sampling methods are all invasive, thus it would be of benefit if a way could be found to assess the endometrium using a relatively non‐invasive approach. Such a technique would need to be relatively easy to learn and perform, as well as being well accepted by the patients. We believe that transvaginal sonography fulfills many of these requirements, the following review will attempt to put forward some of the evidence to support this view.
Changes in leukocyte, granulocyte and lymphocyte counts following antenatal betamethasone administration to pregnant women
Authors: Kadanali S. , Ingeç M. , Küçüközkan T. , Börekçi B. , Kumtepe Y.
Published: July 2002

Objective: Preterm labor and premature rupture of membranes are associated with a mild leukocytosis. However, we have observed a higher maternal leukocyte count after antenatal betamethasone therapy. We planned this study to evaluate the effects of antenatal betamethasone treatment on maternal leukocyte, granulocyte and lymphocyte count. Methods: Forty‐six pregnant women with the diagnosis of preterm labor between 28 and 33 weeks of gestation age received 12 mg betamethasone at a 12‐h interval. The control group consisted of 50 pregnant women between 28 and 33 weeks of gestational age with no medical or obstetrics problems. After a baseline venous sampling, serial leukocyte, granulocyte and lymphocyte counts were obtained every 6 h until it returned to baseline value. Results: There were no statistically significant differences in the control group with respect to the total leukocyte, lymphocyte, and neutrophil count. Total leukocyte and granulocyte counts were increased by 29.8% and 17.8% within 24 and 12 h after betamethasone injection, respectively (P < 0.01). A significant reduction in lymphocyte count was observed within 12 h (45.4%) after betamethasone injection (P < 0.01). All changes in leukocyte, granulocyte and lymphocyte counts returned to baseline values within 3 days. Conclusion: Antenatal betamethasone therapy leads to an increase in maternal leukocyte count and a decrease in lymphocyte count. This effect is transient and any leukocytosis persisting for more than 3 days is not due to betamethasone administration.
Puerperal tetanus and hysterectomy
Authors: Khajehdehi P. , Rezaian G.R.
Published: July 2002

Objectives: To compare the clinical course and outcome of patients with post‐partum tetanus (n = 21) to those of non‐post‐partum ones (n = 24) and to determine the effect of early hysterectomy on the survival of cases with severe post‐partum tetanus (n = 20). Methods: In this prospective study we compared the clinical course and outcome of post‐partum and non‐post‐partum cases of tetanus admitted to the intensive care unit. Moreover, early hysterectomy was done on every other patient with severe post‐partum tetanus (n = 10). Chi‐square and Student's t‐tests were used for statistical purposes. Results: Post‐partum cases were younger (P = 0.002), had a higher incidence of acute respiratory failure and a mortality rate compared to the non‐post‐partum group. In addition, early hysterectomy did not change the course of severe post‐partum cases. Conclusion: The mortality associated with post‐partum tetanus is comparable to non‐post‐partum cases and early hysterectomy has no effect on its outcome.
The outcome of radiation therapy in elderly patients with advanced cervical cancer
Authors: Minagawa Y. , Kigawa J. , Itamochi H. , Terakawa N.
Published: July 2002

Objective: To clarify whether radiation therapy is a suitable treatment for elderly patients with advanced squamous cell carcinoma of the uterine cervix. Method: We investigated the outcome of 102 patients with stage IIIb squamous cell carcinoma of the uterine cervix, who underwent radiation therapy alone, by using a multiple regression analysis. Results: All factors investigated were not significant prognostic variables in this study. Five‐year survival rates between the two groups, which we have divided for every 5 years of age, showed no differences. Conclusion: Age was not a dependent prognostic factor in the patients with stage IIIb squamous cell carcinoma of the uterine cervix, if the subject underwent complete treatment. This study suggests that radiation therapy is still considered as a reasonable treatment for elderly patients with cervical cancer.
Gestational diabetes mellitus and glucose intolerance in a Mexican population
Authors: Forsbach G , Cantú‐Diaz C , Vázquez‐Lara J , Villanueva‐Cuellar M.A , Alvarez y Garcı́a C , Rodrı́guez‐Ramı́rez E
Published: July 2002

Objective: To determine the prevalence of gestational diabetes mellitus (GDM), glucose intolerance (GI) and macrosomia in a Latin Mexican population, using the one‐step procedure proposed by the World Health Organization. Method: Consecutive pregnant women (n=667) who attended the antenatal clinic and delivered at the hospital were recruited after 24 weeks of gestation, mean 34.2 weeks, range 24–40 weeks. After a minimum of 3 days with a 150‐g carbohydrate diet, a glucose load of 75 g was administered and 2 h after, a blood sample for glucose was taken, plasma was separated and immediately processed for glucose, using a glucose‐oxidase method. Results: Twenty women were diagnosed with GDM (3%), 87 had GI (13%) and 560 had a normal glucose response (84%). Macrosomia occurred in two (10%) of GDM, in nine (10.3%) of GI, and in 29 (5.4%) gravidas with a normal response, respectively. Only one of the macrosomic infants whose mother had GI, had neonatal hypoglycemia and hyperbilirubinemia. Conclusion: The overall prevalence of glucose abnormalities during pregnancy was 16%. GI in this group was associated to macrosomia at the same rate as GDM.
Understanding sexual and reproductive violence: an overview
Authors: Barzelatto J.
Published: July 2002

International agreements recognizing different forms of violence as violations of human rights and the definition provided by the 1993 UN Declaration on the Elimination of Violence against Women are taken as a starting point and its implications analyzed, emphasizing gender roles and stereotypes. Violence against women is related to violence in general, to the so‐called culture of violence. Factors influencing a culture of violence are discussed, as well as the differences between public and private violence, emphasizing the need to understand their interaction to be effective in preventing violence against women. It is concluded that all violence stems from unbalanced exercise of power, creating injustice and lack of real democratic interaction. When left unchallenged such situations become part of the culture of individuals and societies, reinforcing the use of violence to solve conflicts. Hence, preventing violence against women requires cultural, social, economic, and political changes that are only possible by mobilizing society as a whole.