bluuebunny/crossref_metadata_2025 huggingface dataset:
Yield and Yield Attributing Parameters of Toria (Brassica campestries) under Real Time Rainfall Situation in an Inceptisols of Assam, India
Authors: Borah R. , Baruah N. , Sarma P. K. , Borah R. , Sonowal A. , Borah P. , Kalita R. , Borkotoki B. , Neog P.
Published: January 2021

A field experiment was conducted during rabi season of 2018-19 and 2019-20 in Dryland experimental field belong to soil order Inceptisols, Biswanath college of Agriculture, Assam Agricultural University, Biswanath chariali, Assam to study the ‘‘Yield and yield attributing parameters of toria (Brassica campestries) under real time rainfall situation in an Inceptisols of Assam, India’’ under AICRPDA, NICRA. The treatments consisting of 4 different dates of sowing i.e. S1-41th SMW, S2-44th SMW, S3-46th SMW, and S4- 48th SMW, & three variety i.e. V1-JT-90-1(Jeuti), V2-Yellow sarson (Benoy) and V3- TS-38. Growth, yield and yield attributing characters of toria varieties were influenced by different dates of sowing. S1 registered higher plant height (43.2 cm, 92.9 cm and 106.6 cm & 40.2 cm, 89.8 cm and 101.5 cm) and number of branch (3.8, 5.3 and 7.2 & 3.4, 5.1 and 6.9) at 30 DAS, 45 DAS and 60 DAS, respectively, during 2018-19 and 2019-20. Yield attributing characters like number of siliqua, number of seed per siliqua, 1000 seed weight (g) were gradually decreased with advancement of sowing dates. Among the three varieties V1 (Jeuti) recorded highest seed yield (8.9 q ha-1 and 8.1 q ha-1) and stover yield (23.4 q ha-1 and 22.2 q ha-1) in 2018-19 and 2019-20, respectively. Highest HI (28.5% and 25.8%) was recorded in S1 and lowest was recorded in S4 (20.7% and 14.6%).
As Climate Change Responds with Terrifying Brutalities
Authors: MRICS Nura Jibo
Published: January 2021

Introduction: The lip service in tackling the climate change issues five years after the famous Paris Agreement on climate change is quite unwholesome to individual countries' pledges and promises that were made on reducing global carbon emissions at Le Bourget, France. The attempt to limit the global mean temperature to 1.50 Celsius preindustrial level has even resulted in warming the climate more than anticipated [1]. The bulk of the climate change adaptation and mitigation effort(s) have, generally, ended up in a tragic fiasco. The rise in sea level and temperature overshoot carry substantial and enormous risks and uncertainties that have caused the entire humanity to head towards an irreversible crossing tipping point [2].  For example, the year 2020 horrible flooding; animal and plant species extinction; coral reef death; permafrost melt; loss of sea and land ice; breaking of the two major glaciers in the coast of the Amundsen Sea in West Antarctica that has kept climatologists and meteorologists terrified in studying the Pine Island and Thwaites glaciers, are all cases in point that proved an irreversible condition of the climate change. 

Methodology: This paper used content analyses by reviewing the climate change brutal scenarios that occurred at random globally. The data on climate events was obtained from the existing literature on the magnitude of destruction of flood rains and storms’ damage due to sea level rise that is exacerbated by the breaking away of the two major glaciers in the coast of the Amundsen Sea in West Antarctica.

Results: The paper examined critically the flood scenarios that occurred in Puerto Rico; Arecibo; Panzhihua; Gopalganj; Kara-Kache; Krasnoyarsk; Tlalpane; Talas; Taif; Valencia; Lagamenas; Khabarovsk; Hadejia; Dabi; Magarya; and Auyo etc. It revealed that massive flooding was witnessed globally within a span of a weeklong catastrophe. Sea level had risen by at least 0.05 percentile as a result of the breaking away and melting of the two glaciers at the coast of the West Antarctica.

Conclusion: The paper concluded that human beings are no longer near the target of achieving the 1.50–20C goal. What remains now is for everyone to understand the dangers of the climate change blind investment that has already thrown the entire world habitat into a déjàvu phenomenon.
Menkes Disease after Copper Histidine Replacement Therapy: Case Report
Authors: George David H. , Casey Robin E.
Published: July 2002

Menkes disease (MD) is an X-linked recessive disorder of copper metabolism, characterized in its untreated state by progressive disorders of multiple systems, especially the central nervous system (CNS) and connective tissue, and death by 3 years of age. Recently, therapy with copper-histidine has modified the severity of MD and permitted survival into adolescence. Clinical response has been greater for the neurological abnormalities than for the connective tissue abnormalities. In this report, we describe the postmortem pathology of one individual who had received copper-histidine therapy and died at age 10; we believe this to be the first such pathological report. The postmortem examination demonstrated significant pathology of mesenchymal tissues, including skeletal abnormalities, vascular degeneration, and bladder diverticula. The CNS, by contrast, showed minimal pathology. The phenotype was more consistent with occipital horn syndrome, a milder allelic disorder of copper metabolism, than with classic MD. The differential sensitivity of CNS and mesenchymal tissues to copper-histidine therapy may result from heterogeneity in the response of different copper-dependent enzymes.
Polysome‐ribosome distribution in isogenic RCstr and RCrel strains of Escherichia coli
Authors: Matzura H.
Published: July 2002

Study of the relative proportions of ribosomes and polysomes released by a standardized lysing procedure from isogenic RCstr and RCrel strains of Escherichia coli shows that a 20‐min period of amino acid starvation of RCstr bacteria reduces the fraction of ribosomes recovered in polysomes to about 60% of its value characteristic of exponentially growing cells: A similar starvation treatment of the RCrel bacteria causes no appreciable reduction in the fraction of polysomal ribosomes.
Antigenic determinants of two components of proteoglycan complex from bovine cartilage
Authors: Di Ferrante Nicola , Donnelly Patricia V. , Gregory John D. , Sajdera Stanley W.
Published: July 2002

The immunological properties of a glycoprotein fraction and of proteoglycan subunits obtained from bovine nasal cartilage by nondisruptive methods of isolation have been studied. Using the techniques of hemagglutination and hemagglutination inhibition, we found that the glycoprotein contains most of the species‐specific determinants, whereas the proteoglycan subunits contain most of the cross‐reacting ones.
Localisation de l'activite phospholipasique dans les tissus vegetaux
Authors: Clermont H. , Douce R.
Published: July 2002

Mitochondria and plastids isolated from various plant tissues by methods which exclude contamination by other cell organelles lack phospholipase D activity.
The effect of a purified preparation of lecithinase C on iodide transport
Authors: Macchia V. , Wolff J.
Published: July 2002

Treatment of porcine thyroid slices with highly purified lecithinase C leads to a marked reduction in the ability to accumulate iodide ion. Although this response is less sensitive as a function of lecithinase concentration than is the ability to respond to thyrotropin with an increase in glucose oxidation, phospholipid synthesis or adenyl cyclase activity, it occurs when the baseline values of these parameters are not substantially altered.
Seminalplasmin inhibits transcription and translation of φ80 DNA in vitro
Authors: Rao Veena N. , Reddy E.S.P. , Torriani Annamaria , Bhargava P.M.
Published: July 2002

Seminalplasmin, an antimicrobial protein from bovine seminal plasma that has been earlier shown to inhibit transcription in whole cells and by purified RNA polymerase in vitro, but not translation in whole cells, is now shown to inhibit both transcription and translation independently of each other, in a coupled transcription‐translation system from E. coli using φ80dphoAlacZ DNA as the template.
High affinity proline uptake in rat brain synaptosomes
Authors: Hauptmann Miroslawa , Wilson David F. , Erecińska Maria
Published: July 2002

The uptake of L‐proline by synaptosomes isolated from different regions of the brain was investigated. The highest rates of transport and the largest accumulation ratios were found in synaptosomes from the midbrain, striatum, hippocampus and hypothalamus, with lower activity in the cortex and medulla (+ pons) and lowest in the cerebellum. The high affinity, Na+ ‐dependent proline uptake had a K
m of 12 μM, a V
max of 0.6 nmol.min−1.mg protein−1 and the maximum accumulation ratio ([proline]i/[proline]o) at 2 μM added radioactive amino acid was 40–50. Our results suggest that: (1) only 5–10% of the synaptosomal population accumulates proline through the high‐affinity uptake; (2) the characteristics of the proline uptake system into the prolinergic nerve endings are remarkably similar to those of other amino acid neurotransmitters (GABA, aspartate and glutamate); (3) the behavior of proline is consistent with this amino acid being a neurotransmitter in the central nervous system.
Amino acid sequence of human factor D of the complement system
Authors: Johnson D.M.A. , Gagnon J. , Reid K.B.M.
Published: July 2002

The amino acid sequence of human factor D is proposed from the analysis of the peptides produced by treatment of the factor D with cyanogen bromide, iodosobenzoic acid, trypsin and V‐8 protease. Comparison of the proposed sequence with the sequences of other serine esterases indicated that factor D, although it is a plasma serine esterase, is more closely related to certain proteases not found in the plasma than to other plasma serine esterases of the complement system. For example, 36% and 32% identity in amino acid sequence was found on comparison of factor D with elastase and group‐specific protease, respectively. Whereas only 27% and 18% identity was observed between factor D and the other complement serine esterases, Clr and factor B, respectively.
Oscillation of delayed luminescence from PS II: recombination of S2Q−B and S3Q−B
Authors: Rutherford A.W. , Inoue Y.
Published: July 2002

Luminescence decaying in the seconds to minutes time scale was studied in spinach chloroplasts and the following results were obtained: (1) After a series of flashes a slow phase which decays in the tens of seconds to minutes time scale was observed to oscillate with a pattern characteristic of S2Q−
B and S3Q−
B recombination. This phase was lost upon Tris‐treatment or upon the addition of DCMU. (2) After every flash a small faster phase of luminescence decaying in the seconds time scale was also present. This phase progressively increased with increasing numbers of flashes but when methyl viologen was present no such progressive increase of this phase occurred. In the presence of DCMU this seconds time scale luminescence was greatly increased. This phase of luminescence is attributed to S2Q−
A recombination. (3) Tris‐treatment resulted in the appearance of an even faster phase of luminescence which may be due to Z+Q−
B recombination. These results demonstrate a close correlation of the kinetics of luminescence decay with thermoluminescence emission temperature.
Inhibition of synaptosomal 5‐[3H]hydroxytryptamine uptake by endogenous factor(s) in human blood
Authors: Angel Itzchak , Paul Steven M.
Published: July 2002

The inhibition of 5‐[3H] hydroxytryptamine uptake into rat forebrain synaptosomes by human plasma extracts was studied. Highly potent, small (M
r < 10000), and heat stable factor(s) were found to inhibit 5‐[3H] hydroxytryptamine uptake specifically, reversibly and in a non‐competitive manner. The possible role of these factor(s) as endogenous modulators of serotonergic activity is discussed.
Anti‐LPS factor in the horseshoe crab, Tachypleus tridentatus
Authors: Ohashi Kunihiro , Niwa Makoto , Nakamura Takanori , Morita Takashi , Iwanaga Sadaaki
Published: July 2002

Anti‐LPS factor, which inhibits the endotoxin mediated coagulation system in the horseshoe crab, Tachypleus tridentatus, was found to lyse red blood cells sensitized with gram‐negative bacterial LPS, but not to lyse unsensitized cells. This hemolysis occurred even at 0°C and was completed within 1 min. The binding of anti‐LPS factor to LPS must be essential for the hemolysis, because free LPS inhibited the hemolytic action of anti‐LPS factor.
Species variations amongst lysosomal cysteine proteinases
Authors: Kirschke Heidrun , Ločnikar Pavel , Turk Vito
Published: July 2002

Properties of cathepsin L from rat liver lysosomes were compared with those of a similar enzyme, cathepsin S from beef spleen. Major characteristics of cathepsin L are the high activity against Z‐Phe‐Arg‐methylcoumarylamide and sensitivity to the fast reacting irreversible inhibitor Z‐Phe‐Phe‐diazomethane. In contrast, cathepsin S hydrolyzes Z‐Phe‐Arg‐methylcoumarylamide only slowly and Z‐Phe‐Phe‐diazomethane cannot be regarded as a potent inhibitor of this enzyme. The differences in the substrate specificity of cathepsin L from rat liver and cathepsin S from beef spleen are discussed in comparison with the substrate specificity of cathepsin B from rat and human liver and beef spleen.
Conformations and CD curves of cyclo(L‐ or D‐Phe‐L‐Pro‐Aca): cyclized models for specific types of β‐bends
Authors: Lee Sannamu , Mizuno Hiroshige , Nakamura Hideaki , Kodera Yasushi , Kato Tetsuo , Gō Nobuhiro , Izumiya Nobuo
Published: July 2002

β‐Bend
CD Measurement
Conformational energy calculation
Cyclic peptide
Gramicidin S
NMR measurement
Degradation of oxalate in rats implanted with immobilized oxalate oxidase
Authors: Raghavan K.G. , Tarachand U.
Published: July 2002

Accumulation of oxalate leads to hyperoxaluria and calcium oxalate nephrolithiasis in man. Since oxalate is a metabolic end product in mammals, the feasibility of its enzymic degradation has been tested in vivo in rats by administering exogenous oxalate oxidase. Oxalate oxidase, isolated from banana fruit peels, in its native form was found to be non‐active at the physiological pH of the recipient animal. However, its functional viability in the recipient animal was ensured by its prior binding with ethylenemaleic anhydride, thus shifting its pH activity curve towards the alkaline range. Rats implanted with dialysis membrane capsules containing such immobilized oxalate oxidase in their peritoneal cavities effectively metabolized intraperitoneally injected [14C]oxalate as well as its precursor [14C]glyoxalate. The implantation of capsules containing coentrapped multienzyme preparations of oxalate oxidase, catalase and peroxidase led to a further degradation of administered [14C]oxalate in rats.
Complete amino acid sequence of alkaline mesentericopeptidase
Authors: Svendsen Ib , Genov Nicolay , Idakieva Krassimira
Published: July 2002

This paper reports the complete amino acid sequence of a novel subtilisin: alkaline mesentericopeptidase. The protein contains 275 amino acid residues in a single polypeptide chain. The sequence data presented allow the linear arrangement of the whole molecule. Comparison with the subtilisins amylosacchariticus, novo, carlsberg, DY, and a subtilisin isolated from B. subtilis strain I 168, respectively classifies the enzyme as a serine protease of the subtilisin novo type closely related to subtilisin amylosacchariticus. Special features of mensentericopeptidase with respect to enzymatic and physico‐chemical properties are discussed.
Purification of ovine placental lactogen (oPL) using high‐performance liquid chromatography
Authors: Chan John S.D. , Nie Z.-R. , Seidah N.G. , Chrétien M.
Published: July 2002

After initial purification of ovine placental lactogen (oPL) using the procedures described previously [(1976) Endocrinology 98, 65‐75], the oPL preparation was further purified by high‐performance liquid chromatography (HPLC) using an anionic exchange column (Bio‐Sil TSK DEAE‐2‐SW). Two forms of oPL with different relative mobilities on HPLC were isolated and designated oPL‐I and oPL‐II. Subsequent analysis by polyacrylamide gel electrophoresis containing SDS revealed that oPL‐I and oPL‐II are nearly homogeneous (greater than 90% pure) and are identical in apparent M
r (approx. 22 000–23 000). Like human growth hormone (hGH), oPL‐I and oPL‐II are equally active in the radioreceptor assays for growth hormone‐like activity (RRA‐GH) and for prolactin‐like activity (RRA‐RRL). In the radioimmunoassay of oPL, both oPL‐I and oPL‐II are immunologically similar. Analysis of amino acid composition revealed that oPL‐I and oPL‐II consist of 199 and 196 residues, respectively, and have almost identical residues except that oPL‐I has a higher content of glycine. Furthermore, both oPLs have a general similarity in amino acid composition to oGH and oPRL except for a lower content of methionine and leucine but with a higher content of lysine. Our studies demonstrated the presence of two similar forms of oPL. Whether these two similar forms of oPL share identical primary structure remains to be determined.
Chromatographic evidence for high‐molecular‐mass galanin immunoreactivity in pig and cat adrenal glands
Authors: Bauer Friedrich E. , Adrian Thomas E. , Yanaihara Noboru , Polak Julia M. , Bloom Stephen R.
Published: July 2002

Galanin
Molecular form
Adrenal gland
Radioimmunoassay
Chromatography
Complete recovery of the phosphoenzyme‐forming activity of nucleoside‐diphosphate kinases after reconstitution of their subunits
Authors: Yokoyama Minehiko , Uesaka Hiroshi , Ohtsuki Kenzo
Published: July 2002

Two distinct subunits [α‐subunit (M
r 21 000, pI 7.6) and β‐subunit (M
r 19000, pI 6.5)] of nucleoside‐diphosphate (NDP) kinases highly purified from HeLa S3 cells can be separated by FPLC using a Mono P column in the presence of 6 M urea and 1% pharmalyte (pH range between 5.0 and 8.0). Comparatively high [32P]‐phosphate incorporation was detected when these two subunit fractions were reconstituted in vitro. Available evidence suggests that these two enzyme subunits are necessary for the formation of phosphoenzyme, which functions as an intermediate in NDP kinase action
Preliminary X‐ray studies of the tetra‐heme cytochrome c3 and the octa‐heme cytochrome c3 from Desulfovibrio gigas
Authors: Sieker L.C. , Jensen L.H. , LeGall J.
Published: July 2002

A tetra‐heme and an octa‐heme cytochrome c
3 from the sulfate bacterium Desulfovibrio gigas have been crystallized. Diffraction quality crystals of the tetra‐heme cytochrome are obtained from solution by the addition of polyethylene glycol at pH 6.5. The crystals are orthorhombic, space group P21212 with unit cell parameters a = 42.27 Å, b = 52.54 Å and c = 52.83 Å. The octa‐heme cytochrome crystals develop from low ionic strength solutions of phosphate or Tris‐Cl in the pH range 6.2–7.6. The crystals belong to the trigonal system, space group P31 or the enantiomorph P32, with unit cell parameters a = b = 57.4 Å, c = 97.3 Å, γ = 120°. Single crystal diffraction studies of the structures of these two low‐potential cytochromes are in progress.
A common bicyclic protein kinase cascade inactivates the regulatory enzymes of fatty acid and cholesterol biosynthesis
Authors: Carling David , Zammit Victor A. , Hardie D.Grahame
Published: July 2002

A highly purified rat liver protein kinase phosphorylates and inactivates acetyl‐CoA carboxylase, and causes rapid inactivation of microsomal HMG‐CoA reductase in the presence of MgATP. Both effects are stimulated in an identical manner by AMP, and are greatly reduced by prior treatment of the kinase with purified protein phosphatase. The dephosphorylated kinase can be reactivated in the presence of MgATP, apparently due to a distinct kinase kinase, and this reactivation is stimulated by nanomolar concentrations of palmitoyl‐CoA. These results show that a common, bicyclic protein kinase cascade can potently inactivate the regulatory enzymes of both fatty acid and cholesterol biosynthesis.
Glutamate acting on NMDA receptors stimulates neurite outgrowth from cerebellar granule cells
Authors: Pearce Ian A. , Cambray-Deakin Martin A. , Burgoyne Robert D.
Published: July 2002

The effect of endogenous glutamate on neurite outgrowth from cerebellar granule cells in culture was examined. Neurite outgrowth was inhibited by enzymatic removal of endogenous glutamate from the culture medium. The broad‐spectrum glutamate receptor antagonist kynurenate also inhibited neurite outgrowth from granule cells in serum‐containing and serum‐free cultures; the inhibition by kynurenate was reversed by exogenous glutamate. Neurite outgrowth was inhibited to the same extent by the NMDA receptor antagonist APV. These results indicate that endogenous glutamate, possibly released by granule cells themselves, stimulated neurite outgrowth through activation of the NMDA class of glutamate receptors. Activation of NMDA receptors on developing neurons may be an important mechanism for the regulation of neuronal growth and differentiation.
Inositol phospholipids are probably not the source of arachidonic acid for eicosanoid synthesis in astrocytes
Authors: Pearce Brian , Jeremy Jamie , Morrow Christine , Murphy Sean , Dandona Paresh
Published: July 2002

In astrocyte‐enriched cultures of the rat cerebral cortex the Ca2+ ionophore A23187 provoked the breakdown of inositol phospholipids, the liberation of arachidonic acid and the release of prostaglandins E2, F2α, I2 and thromboxane A2. However, agonists for receptors also coupled to inositol phospholipid metabolism in these cells failed to produce an increase in the release of both arachidonic acid and eicosanoids. Results suggest that the A23187‐stimulated release of arachidonic acid and eicosanoids is caused by a phospholipase A2‐mediated attack on lipids other than the inositol phospholipids. Moreover, receptors linked to inositol lipid turnover are not involved in the control of eicosanoid release from astrocytes.
Inhibition of actin polymerization by latrunculin A
Authors: Coué Martine , Brenner Stephen L. , Spector Ilan , Korn Edward D.
Published: July 2002

Latrunculin A, a toxin purified from the red sea sponge Latrunculia magnifica, was found previously to induce striking reversible changes in the morphology of mammalian cells in culture and to disrupt the organization of their microfilaments. We now provide evidence that latrunculin A affects the polymerization of pureactin in vitro in a manner consistent with the formation of a 1:1 molar complex between latrunculin A and G‐actin. The equilibrium dissociation constant (K
d) for the reaction in vitro is about 0.2 μM whereas the effects of the drug on cultured cells are detectable at concentrations in the medium of 0.1–1 μM.
Different effects of two thromboxane A2/prostaglandin H2 receptor ligands, U46619 and S‐145, on rabbit platelets
Authors: Nakano Tohru , Hanasaki Kohji , Arita Hitoshi
Published: July 2002

Stimulation of rabbit platelets with U46619 induced platelet shape change, aggregation and secretion of ATP. However, S‐145, which specifically binds to the thromboxane A2/prostaglandin H2 receptor like U46619, induced only shape change. Both compounds rapidly elevated cytoplasmic Ca2+ concentration although only U46619 evoked the formation of inositol phosphates. Chelating external Ca2+ with EGTA did not affect the S‐145‐induced platelet shape change while intracellular Ca2+ movement was severely reduced. These results suggest an essential role of phospholipase C in the induction of platelet aggregation and secretion and that some factor other than Ca2+ and phospholipase C participates in platelet shape change.
The β‐bungarotoxin‐binding protein from chick brain: binding sites for different neuronal K+ channel ligands co‐fractionate upon partial purification
Authors: Schmidt Ralf R. , Betz Heinrich
Published: July 2002

β‐Bungarotoxin (β‐Butx) is a presynaptically active neurotoxin which blocks neuronal A‐type K+ channels. Here, the efficient solubilisation and about 300‐fold purification of the β‐Butx‐binding protein from chick brain were achieved by detergent extraction at high ionic strength followed by chromatography on DEAE Affigel Blue, β‐Butx Affigel 102 and wheat germ agglutinin Sepharose. Binding of 125I‐labelled β‐Butx to the purified protein was inhibited by two other K+ channel ligands, dendrotoxin I and mast cell‐degranulating peptide. It is concluded that the β‐Butx‐binding protein is a member of a family of voltage‐gated K+ channels which exhibit varying affinities for different polypeptide ligands.
ATP:AMP phosphotransferase activity, a new characteristic of Catharanthus roseus tonoplasts
Authors: Hill M. , Dupaix A. , Nhiri M. , Guyen L. , Arrio B.
Published: July 2002

The ATPase activity of Catharanthus roseus tonoplasts was examined using HPLC separation and quantification of adenine nucleotides. ATP seemed to be degraded into ADP and AMP by tonoplast vesicles. When ADP was the initial substrate, the appearance of AMP and concomitant ATP synthesis were observed; these reactions were inhibited by Ap5A. The apparent degradation of ATP into AMP was also inhibited by Ap5A. These results indicated that AMP arose from an ATP:AMP phosphotransferase activity and excluded the possibility of the hydrolysis of ADP into AMP by the tonoplast ATPase. AMP was degraded by the microsomal fraction from protoplasts or by the cytosol while the tonoplast vesicles did not hydrolyze it. This observation was used to assess the purity of tonoplasts.
A pertussis toxin‐sensitive GTP‐binding protein plays a role in the G0‐G1 transition of rat hepatocytes following establishment in primary culture
Authors: Fujinaga Yukako , Morozumi Naomi , Sato Koichi , Tokumitsu Yukiko , Fujinaga Kei , Kondo Yoicho , Ui Michio , Okajima Fumikazu
Published: July 2002

Acute spontaneous c‐myc gene expression and sustained increase of a GTP‐binding protein(s) (G‐protein) which is sensitive to islet‐activating protein (IAP), pertussis toxin, occurred early during primary culture of adult rat hepatocytes. Following these earlier events, DNA synthesis was demonstrated in response to EGF and insulin. Addition of IAP immediately after plating of primary cultures inhibited c‐myc expression and the hormone‐induced DNA synthesis. Addition at 24 h or later following cell inoculation, however, produced only weak effects on DNA synthesis, even though the IAP‐sensitive G‐proteins were completely inactivated. We conclude that the IAP‐sensitive G‐protein(s) plays a role in the earlier process(es) of the G0‐G1 transition, which is essential for the initiation of growth factor‐dependent DNA synthesis.
Expression of RNA transcripts for the postsynaptic 43 kDa protein in innervated and denervated rat skeletal muscle
Authors: Froehner Stanley C.
Published: July 2002

A cDNA clone encoding the mouse muscle postsynaptic 43 kDa protein was isolated and sequenced. The amino acid sequence of this protein, which is closely associated with nicotinic acetylcholine receptors at Torpedo electrocyte and vertebrate skeletal muscle synapses, is very similar in different species. A cysteine‐rich region homologous to part of the regulatory domain of protein kinase C may be important in interactions of this protein with the lipid bilayer. RNA transcripts for the 43 kDa protein increase only 2–3‐fold after denervation of rat skeletal muscle, in sharp contrast to the α‐subunit of the muscle nicotinic receptor which increases more than 30‐fold. Thus, the expression of these two proteins is regulated by different mechanisms.
Purification and reconstitution of potassium channel proteins from squid axon membranes
Authors: Prestipino G. , Valdivia H.H. , Liévano A. , Darszon A. , Ramírez A.N. , Possani L.D.
Published: July 2002

Voltage‐dependent K+ channels are responsible for repolarization of the cell membrane during the late phase of the action potential. Here we report the purification of proteins from squid axon membranes which bind the K+‐channel blocker noxiustoxin (NTX), and their subsequent functional reconstitution in planar bilayers. The NXT‐affinity purified proteins had Mr values of 60000 ± 6000, 160000 ± 15000 and 220000 ± 20000. Their incorporation into bilayers resulted in single‐channel currents with three conductances, the most frequent one of 11 pS in 300/100 mM KCl (cis/trans). The voltage dependence, reversal potential and bursting behavior suggest that these are the K+ channels involved in the squid axon action potential.
Maximal activities of glutaminase, citrate synthase, hexokinase, phosphofructokinase and lactate dehydrogenase in skin of rats and mice at different ages
Authors: Keast David , Nguyen Tam , Newsholme Eric A.
Published: July 2002

The activities of key glycolytic enzymes are, in general, similar at the three different ages of the animals used in this work (very young, adult and old). Glutaminase is present in skin of both mice and rats, but the activity was much lower in adult animals compared to the very young or the old. It is suggested that this activity is important for the provision of nitrogen for the de novo synthesis of purine and pyrimidine nucleotides during the growth of skin in the young animal and for DNA repair in the old animals; it might be important in the adult skin in response to wound healing.
Is the pseudo‐dyad in retroviral proteinase monomers structural or evolutionary?
Authors: Rao J.K.Mohana , Wlodawer Alexander
Published: July 2002

A pseudo‐dyad was found to exist in the monomers of the crystal structures of the proteinases from Rous sarcoma virus and the human immunodeficiency virus. This dyad, also discovered earlier in pepsin‐like aspartic proteinases and considered to be of probable evolutionary origin, has been shown to arise as a result of the topology and the folding of the proteinase monomers and may not therefore have much evolutionary significance.
Agonist‐induced down‐regulation of m1 muscarinic receptors and reduction of their mRNA level in a transfected cell line
Authors: Wang Shou-Zhen , Hu Jingru , Long Rochelle M. , Pou Wanida S. , Forray Carlos , El-Fakahany Esam E.
Published: July 2002

Agonist‐induced reduction in both the number of ml muscarinic receptors and the mRNA coding for the receptor protein was investigated in Chinese hamster ovary (CHO) cells which were transfected with the ml muscarinic receptor gene. Receptor concentration was measured by the specific binding of the muscarinic ligand, [3H]quinuclidinyl benzilate ([3H]QNB), and Northern blot hybridization analysis was used to evaluate the levels of receptor mRNA. Incubation of cells with 1 mM of the muscarinic agonist, carbamylcholine (CBC), for 24 h decreased receptor density and mRNA levels in cells by 65% and 73%, respectively. These results indicate that agonist‐induced down‐regulation of ml muscarinic receptors might be due to, at least in part, a decrease in receptor synthesis resulting from a reduction in the steady‐state level of their mRNA.
Ligustrazine selectively blocks ATP‐sensitive K+ channels in mouse pancreatic β cells
Authors: Peers C. , Smith P.A. , Nye P.C.G.
Published: July 2002

Ligustrazine potently inhibits hypoxic pulmonary vasoconstriction. This effect can be mimicked by the sulphonylurea tolbutamide, a blocker of ATP‐sensitive K+ channels. Here we demonstrate that ligustrazine is a specific blocker of ATP‐sensitive K+ channels in mouse pancreatic β cells. This suggests that ligustrazine may act in the pulmonary circulation by blocking ATP‐sensitive K+ channels.
Oligo(3'‐deoxyADP‐ribosyl)ation of thenuclear matrix lamins from rat liver utilizing 3'‐deoxyNAD as a substrate
Authors: Pedraza-Reyes Mario , Alvarez-Gonzalez Rafael
Published: July 2002

It has previously been shown that the levels of poly(ADP‐ribose)polymerase and polymers of ADP‐ribose that co‐purify with the nuclear matrix in regenerating liver fluctuate with the levels of in vivo DNA replication [(1988) FEBS Lett. 236, 362‐366]. We have now electrophoretically identified lamins A and C, and poly(ADP‐ribose)polymerase as the main protein targets for poly(ADP‐ribosyl)ation in isolated nuclear matrices from adult rat liver. The identification of these protein acceptors was facilitated by the utilization of 32P‐radiolabeled 3'‐deoxyNAD as a substrate for nuclear matrix extracts in the presence of exogenously added DNA‐dependent poly(ADP‐ribose)polymerase from calf thymus. The extent of protein modification was time‐ and substrate concentration‐dependent. These results are consistent with the hypothesis that the poly(ADP‐ribose) modification of the lamins A and C and poly(ADP‐ribose)polymerase are important to modulate chromatin‐nuclear matrix interactions in rat liver.
Molecular cloning and sequence analysis of cDNA encoding human cholesterol 7α‐hydroxylase
Authors: Noshiro Mitsuhide , Okuda Kyuichiro
Published: July 2002

A complete cDNA clone encoding human cholesterol 7α‐hydroxylase has been isolated using a rat P‐450ch7α cDNA insert [(1989) FEBS Lett. 257, 97‐100] as a probe and totally sequenced. The cDNA contained 1512‐base pair open reading frame encoding 504 amino acid residues (M
r, 57630), 39‐base pair 5'‐untranslated region 1322‐base pair 3'‐untranslated region including 20 nucleotides of poly A tail in the total length of 2873 base pairs. The deduced amino acid sequence showed 82% similarity to rat P‐450ch7α. Unique amino acid residues were observed in putative binding domains for heme and steroid which are highly conserved in most steroidogenic P‐450s.
mRNA acetylated at 2'‐OH‐groups of ribose residues is functionally active in the cell‐free translation system from wheat embryos
Authors: Ovodov S.Yu. , Alakhov Yu.B.
Published: July 2002

Modification (acetylation) of 2'‐OH groups of mRNA ribose residues does not result in a loss of their template activity in a cell‐free translation system from wheat embryos and in some cases activates inactive mRNAs. Modification of 70–75% of 2'‐OH groups makes mRNA stable to the effect of pancreatic ribonuclease.
Central helix role in the contraction‐relaxation switching mechanisms of permeabilized skeletal and smooth muscles with genetic manipulation of calmodulin
Authors: Gulati Jagdish , Persechini Anthony , Babu Árvind
Published: July 2002

A prominent common feature of calmodulin and troponin structures is the unusually long central helix which separates the two lobes, each containing two Ca2+‐binding sites. To study the role of certain highly conserved residues in the helix in the contraction‐relaxation switching mechanism in muscle, we measured the Ca2+‐activated force of permeabilized skeletal and smooth muscles with three genetically manipulated forms of calmodulin. Mutated calmodulin was made to substitute for troponin‐C in vertebrate skeletal fiber. The mutants had 1–4 deletions in the conserved cluster (positions 81–84) in the solvent‐exposed region of the central helix, which also substantially shortened the helix. The force of the maximally activated fiber was found to be diminished only with the mutant in which the entire cluster Ser‐81 to Glu‐84 (CaMΔ81–84) was deleted. All such deletions were found to be completely ineffective in blocking the Ca2+‐switching process in smooth muscle strips. The results show for the first time that at least a part of the highly conserved four‐residue cluster in the central helix is critical for the contraction mechanism of striated muscle. Further, the possibility is raised that the reduced length of the central helix may be a determining factor in the Ca2+‐switching mechanism in fast‐twitch muscle. These findings combined with the results on smooth muscle indicate diversity in the structure‐function specifications for the central helix of calmodulin for different target proteins.
Expression of protein kinase C‐alpha (PKC‐α) and MYCN mRNAs in human neuroblastoma cells and modulation during morphological differentiation induced by retinoic acid
Authors: Tonini Gian Paolo , Parodi Maria Teresa , Di Martino Daniela , Varesio Luigi
Published: July 2002

It is known that PKC is differently expressed in brain and the peripheral nervous system and is involved in cellular differentiation. We have analyzed 9 human neuronal‐derived crest‐cell lines for PKC‐α mRNA. Seven out of nine expressed 9.0 kb and 4.0 kb PKC‐α mRNAs but three had high level of 9.0 kb transcription. The different expression of the two messenger RNAs may result from alternative splicing and a different degree of cell maturation. The same cell lines were studied for MYCN gene expression. A possible relation between the two genes is discussed. One cell line expressing high levels of both PKC‐α mRNA was treated with 10−1 M retinoic acid (RA). The expression of both messenger RNAs was suppressed when the cells achieved a morphological differentiation and showed neurite‐like processes. A decrease of PKC‐α gene expression was associated to down regulation of MYCN mRNA. These preliminary results suggest that PKC suppression of PKC‐α mRNA is associated with reversion of the malignant phenotype.
Studies of the small heat shock proteins of Caenorhabditis elegans using anti‐peptide antibodies
Authors: Hockertz Michael K. , Clark-Lewis Ian , Candido E. Peter M.
Published: July 2002

Peptides corresponding to selected regions of the 16 kDa small heat shock proteins (hsps) of the nematode C. elegans were synthesized and used to elicit polyclonal antibodies. It was found that these antibodies reacted predominantly with either the 16 kDa or the 18 kDa proteins, suggesting a close structural similarity between these hsps. Western blots of two‐dimensional gels revealed extensive heterogeneity in these proteins, probably resulting from post‐synthetic modifications. The native structures or both size classes or hsps were found to consist of large complexes of 4–5 × 101 Da.
Decreased heat‐ and tumor necrosis factor‐mediated hsp28 phosphorylation in thermotolerant Hela cells
Authors: Arrigo Andre-Patrick , Michel Marcel R.
Published: July 2002

Heat shock or tumor necrosis factor rapidly stimulated the phosphorylation of the mammalian low molecular weight stress protein hsp28. We have found that both phenomena are greatly decreased in cells which are made tolerant to heat. This observation correlated with a better survival of thermotolerant cells exposed to either heat or TNF treatment. The results suggest that the phosphorylation of hsp28 may be linked to the resistance of the cells to the deleterious effects induced by either heat or a mediator of inflammation such as TNF.
Isolation and characterization of polyferredoxin from Methanobacterium thermoautotrophicum The mvhb gene product of the methylviologen‐reducing hydrogenase operon
Authors: Hedderich R. , Albracht S.P.J. , Linder D. , Koch J. , Thauer R.K.
Published: July 2002

The methylviologen‐reducing hydrogenase operon of Methanobacterium thermoautotrophicum contains an open reading frame. mvhB, the product or which was predicted to have a molecular weight of 44 kDa and to contain as many as 48 iron atoms in 12 [4Fe‐4S] clusters, and was therefore suggested to be a polyrerredoxin. We have now, for the first time, isolated this polyfarradoxin. Its identity with the mvhB gene product was evidenced by a comparison of the N‐terminal amino acid sequence. The dark‐brown protein of apparent molecular weight 44 kDa was found to contain 53 mol Fe and 43 mol acid‐labile sulfur per mol. The UV/visible spectrum showed two maxima at 280 nm and 390 nm, and a shoulder at 308 nm. The A
390/A
280 ratio was 0.73. The molar extinction coefficient at 390 nm was 170,000 M−1 cm−1. In the dithionite reduced State the protein displayed an EPR spectrum like that of [4Fe‐4S] clusters. The results indicate that the mvhB gene product is indeed a polyferredoxin.
Heat‐shock responsive elements in the induction of the multidrug resistance gene (MDR1)
Authors: Kioka Noriyuki , Yamano Yoshiaki , Komano Tohru , Ueda Kazumitsu
Published: July 2002

The MDR1 gene, considered to be involved in multidrug resistance of cancer cells, is expressed in liver, kidney, small intestine and the blood‐brain barrier. We investigated MDR1 gene expression in the well‐differentiated hepatoma cell line HepG2 after exposure to several stresses and found that sodium arsenite treatment increased MDR1 gene expression 2.6‐fold. Deletion analysis of the MDR1 promoter indicated that the transcriptional activation after exposure to arsenite depends on a 60‐bp region containing two heat‐shock responsive elements.
Microtubule‐associated protein autophosphorylation alters in vitro microtubule dynamic instability
Authors: Raffaelli Nadia , Yamauchi Paul S. , Purich Daniel L.
Published: July 2002

While phosphorylation of high‐molecular‐weight microtubule‐associated proteins (MAPs) alters the assembly properties of microtubules in vitro, virtually nothing is known about the influence of MAP phosphorylation on the time‐scale of microtubule polymer length redistribution, The latter has been used as an index of microtubule assembly/disassembly turnover as predicted by the dynamic instability model (Mitchison, T.M. and Kirschner, M.W. (1984) Nature 312, 237–242). We have now determined that under conditions leading to the incorporation of 8–10 mol phosphoryl groups per mol MAP‐2 (and about 0.2 mol phosphoryl groups per mol MAP‐1 and tau), we can reproducibly observe significant acceleration in the polymer length redistribution process in a manner consistent will greater microtubule dynamic instability. We have also found that MAP phosphorylation resulted in more extensive release of MAPs from microtubules as a function of increasing salt concentration. These results are consistent with a weakening of MAP‐microtubule interactions upon phosphorylation.
Glycosylation inhibits the interaction of invertase with the chaperone GroEL
Authors: Kern Gunther , Schmidt Marion , Buchner Johannes , Jaenicke Rainer
Published: July 2002

During refolding and reassociation of chemically denatured non‐glycosylated invertase from Saccharomyces cerevisiae, aggregation competes with correct folding, leading to low yields of reactivation (Kern et al. (1992) Protein Sci. 1, 120–131). In the presence of the chaperone GroEL, refolding is completely arrested. This suggests the formation of a stable complex between GroEL and non‐native non‐glycosylated invertase. Addition of MgATP results in a slow release of active invertase from the chaperone complex. When GroEL/ES and MgATP are present during refolding, the final reactivation yield increases from 14% to 36%. In contrast, refolding of the core‐glycosylated and the high‐mannose glycosylated forms of invertase is not arrested by GroEL. Only a short lag phase at the beginning of reactivation and a slightly increased reactivation yield (64% to 86% for core‐glycosylated and 62% to 76% for external invertase) indicate a weak interaction of the glycosylated forms with the chaperone.
A 17.6 kbp region located upstream of the rabbit WAP gene directs high level expression of a functional human protein variant in transgenic mouse milk
Authors: Bischoff Rainer , Degryse Eric , Perraud Frédéric , Dalemans Wilfried , Ali-Hadji Dalila , Thépot Dominique , Devinoy Eve , Houdebine Louis-Marie , Pavirani Andrea
Published: July 2002

We have investigated whether DNA regions present in the rabbit whey acidic protein (WAP) promoter/5′ flanking sequence could potentially confer. in vivo, high level expression or reporter genes. Transgenic mice were generated expressing a variant of human α1‐antitrypsin, which has inhibitory activity against plasma kallikrein under the control of a 17.6 kbp DNA fragment located upstream of the rabbit WAP gene. Up to 10 mg/ml of active and correctly processed recombinant protein were detected in mouse milk, thus suggesting that the far upstream DNA sequences from the rabbit WAP gene might be useful for engineering efficient protein production in the mammary glands or transgenic animals.
Identification of four FGF receptor genes in Medaka fish (Oryzias latipes)
Authors: Emori Yasufumi , Yasuoka Akihito , Saigo Kaoru
Published: July 2002

Four types of cDNA clones encoding tyrosine kinases highly homologous to mammalian fibroblast growth factor receptors (FGF‐R) were isolated from Medaka fish (Oryzias latipes) by the reverse transcription‐polymerase chain reaction. Comparison of the four deduced amino acid sequences with four known mammalian FGF‐Rs indicated that four FGF‐R species corresponding to mammalian FGF‐Rs exist universally in vertebrates including fishes, although FGF‐R4 might have diverged sequences between fishes and mammals. Each of four FGF‐R genes is transcribed to various extents as multiple mRNAs possibly by alternative splicing in adult fishes.
Stimulation of SV40 DNA replication by the human c‐myc enhancer
Authors: Kumano Mariko , Nakagawa Tsutomu , Imamura Yasuhiro , Galli Ivo , Ariga Hiroyoshi , Iguchi-Ariga Sanae M.M.
Published: July 2002

In earlier studies we had shown that a transcriptional enhancer sequence exists about 2 kb upstream of the human c‐myc gene. The core sequence necessary for enhancer activity was defined therein as a 21 bp nucleotide element, which also showed autonomous replicating activity [EMBO J. (1988) 7, 3135–3142; EMBO J. (1989) 8, 4273–4279]. Recently, several reports have substantiated the notion that transcription and replication can be concertedly regulated in a larger number of cases than expected. In this report, we took the simian virus 40 (SV 40) orl/promoter as a model system. The SV40 enhancer is known to enhance transcription from its orlpromoter, but to reduce its replication (probably due to a negative feedback). The SV40 enhancer was replaced by the c‐myc enhancer core in order to see its effect upon SV40 DNA replication and transcription. The results showed that besides stimulating transcription, the c‐myc enhancer promoted SV40 DNA replication in monkey Cost cells. Stimulation was only observed when the c‐myc enhancer was inserted in the ‘up‐to‐down’ orientation to the SV40 promoter. The promoting function of the c‐myc enhancer on DNA replication correlated with the transcriptional activation function, as determined by systematic point mutations introduced within the 21 bp core sequence.
Three‐dimensional structure of (1–36)bacterioopsin in methanol—chloroform mixture and SDS micelles determined by 2D 1H‐NMR spectroscopy
Authors: Pervushin Konstantine V. , Arseniev Alexander S.
Published: July 2002

Spatial structures of proteolytic segment A (sA) of bacterioopsin of H. halobium (residues 1–36) solubilized in a mixture of methanol—chloroform (1:1), 0.1 M LiClO4 organic mixture, or in perdeuterated sodium dodecyl sulfate (SDS) micelles, were determined by 2D 1H‐NMR techniques, 324 and 400 NOESY cross‐peak volumes were measured in NOESY spectra of sA in organic mixture and SDS micelles, respectively. The sA spatial structures were determined by local structure analysis, distance geometry calculation with program DIANA and systematic search for energetically allowed side chain rotamers consistent with NOESY cross‐peak volumes. The structures of sA are similar in both milieus and have the right‐handed α‐helical region from Pro? to Met32 with root mean square deviation (RMSD) of 0.25 Å between backbone heavy atoms and fit well with Pro? to Met32 α‐helical region in electron cryo‐microscopy model of bacteriorhodopsin. The N‐terminal region Ala2‐Gly4 of sA in organic mixture has a fixed structure of two consecutive γ‐turns as 2 * 2?‐helix (RMSD of 0.25 Å) stabilized by the Thr? NH⋯O=C Gln3 and Ile4 NH⋯O=C Ala2 hydrogen bonds while this region in SDS micelles has disordered structure with RMSD of 1.44 Å for backbone heavy atoms. The C‐terminal region Gly33‐Asp? of sA is disordered in both milieus. Torsion angles χ1 of sA were unequivocally determined for 13 (SDS) and 11 (organic mixture) of α‐helical residues and arc identical in both milieus.