bluuebunny/crossref_metadata_2025 huggingface dataset:
The Volume of Iranian Raw Silk Exports in the Safavid Period
Authors: Herzig Edmund M.
Published: July 2007

Pre-nineteenth century Iranian history is short on numbers, or at least on reliable ones. Lack of quantitative data is perhaps the greatest obstacle confronting historians of pre-modern Iranian economy and society, and the Safavid period is scarcely better provided than earlier ages. The Safavid archives were destroyed, the Persian histories provide few hard facts, and the accounts of visiting Europeans, though often full of comment on social and economic matters, are generally unreliable when it comes to quantitative information. Safavid Iran thus presents a depressing contrast to its neighbours to East and West, where the earlier and deeper European penetration has left arich record in the archives of several European countries, while in the case of the Ottoman Empire, the survival of extensive state archives can only remind the historian of Iran of how much was lost through the destruction of their Safavid counterparts.
Three Little Known Types of Tribal Storage Bag
Authors: Tanavoli Parviz
Published: July 2007

In the literature a variety of flatwoven tribal bags are referred to generically as kīsah. In the tribal setting, however, these bags are differentiated according to function, e.g., namakdān, “salt bag“; qāshuqdān, “spoon holder”. This short article introduces three little known types of tribal storage bag: the qāshuqdān; the qalyāndān, “water pipe bag“;and the chupuqdān, “pipe holder”.
Sunni Survival in Safavid Iran: Anti-Sunni Activities during the Reign of Tahmasp I
Authors: Johnson Rosemary Stanfield
Published: July 2007

It has traditionally been taught that Iran, a Sunni polity for many centuries, was converted to Twelver Shi'ism virtually overnight when the armies backing the Safavid house took power in 907/1501. In recent years scholars have begun to question the manner and rapidity of the process of conversion, what it meant to be “Shi'i” or “Sunni” in sixteenth-century Iran, and what, if anything, can be said about Safavid Sunnism.

It has been noted that Sunnism in Iran was phased out only gradually. For example, there are references in the Persian chronicles to persecution of Sunnis as late as 1017/1608. Some sources suggest that Sunni influence persisted at the court of Shah Tahmasp and name prominent Sunnis during his reign. As late as the second decade of the eighteenth century, the conqueror of Isfahan, Mahmud Afghan, attempted to alter the balance of population in the city by relocating 5,000 Sunni families from Hamadan.
Barriers to Skills Development in a Local Construction Labour Market
Authors: Watson Judith , Sharp Graham
Published: July 2007

The literature on the construction industry suggests that the structure of labour markets in that industry sets up barriers to skills development. It is often suggested that ‘employer engagement’, leading to increased ‘buy-in’ into skills development, and investment by employers is the way to overcome these barriers. We present an example from a local labour market in South East England, with reference to an intervention in training (‘Constructing Futures’) that has brought private and public sector actors together. This example shows that employer engagement on its own is not sufficient to overcome the barriers created by the labour market structure. We suggest some essential measures to allow more workers to enter the industry and gain skills and reduce the skill shortages endemic to the industry in South East England.
Bringing cheer to a depressed economy — Economic Development in N. Ireland
Authors: Morrissey Mike , Gaffikin Frank
Published: July 2007

In this, the first of two articles, the authors examine the particularly grim problems of the local economy in N. Ireland. They argue that such problems have to be understood in the context of N. Ireland's distinctive politics and administration. Following an assessment of economic development performance in recent decades, they conclude with a critical review of government strategies for the future. In the second article they intend to examine other local suggestions for economic policy, and to outline a “popular” alternative strategy.
OPERATION GROGCOUNT: THE ROUTINE RECORDING OF ALCOHOL INTAKE AT ROYAL NEWCASTLE HOSPITAL
Authors: O'Neill Pat , Walsh Raoul
Published: July 2007

Operation Grogcount, a scheme which involves the compulsory, routine recording of alcohol intake of all adult in‐patients, was commenced at Royal Newcastle Hospital in 1979. The effect of this scheme on the alcohol recording rate is reviewed and the difficulties encountered outlined. Operation Grogcount has doubled the percentage of files containing a quantitative assessment of alchol consumption. It has also increased the identification rate of hazardous drinkers. Suggestions are made for extending the impact of Operation Grogcount on identified hazardous drinkers.
Community Health Workers and the Drug Problem
Authors: O'Neill Pat
Published: July 2007

Abstract: Denial about the habitual misuse of substances is a standard feature of addictive behaviour. Denial by a collective of addicts when they form a sufficient proportion can then become a societal norm. This societal denial is a major factor in the conspiracy of silence.

Breaching denial is a necessary first step for addicted people seeking change. The community response to addictive behaviour must contain the essential steps as defined in the structured intervention technique (“Alconfrontation”) for the individual. They are: (i) definition and acceptance of the nature of the problem; (ii) education about addictive behaviour and the substances concerned, within a relevant perspective; (iii) clarification of the options and objectives; and (iv) contract making.

It is possible for communities to achieve collective change if enough significant catalytic individuals and agencies strive to do so.

In relation to drink driving and analgesic abuse there have already been significant advances. Cigarette smoking has declined rapidly and this wind of change can be fanned. Armidale in New South wales has a Bicentennial objective to be visibly free of cigarette smoking with its theme “Armidale Leads the Smoke‐Free World”.

Innumerable surveys have confirmed that the major factor in society's problems is substance abuse. What is needed now is a core of catalysts to provoke change by increasing awareness, by appropriate education, by exploring tactics for change and by pushing for commitment and contract for change.
Genetic Variations in Vascular Endothelial Growth Factor and Endothelial Nitric Oxide Synthase and Their Contributions to Human Disease
Authors: GERRITSEN MARY E.
Published: July 2007

This review addresses a rapidly growing area of vascular biology, i.e. genomic variations in vascular genes that underlie different human phenotypes. Two of the most important molecular in vascular biology, endothelial nitric oxide synthase (eNOS) and vascular endothelial cell growth factor (VEGF) are discussed. Variations in the eNOS gene have been correlated with a number of human diseases including hypertension, coronary vasospasm, smoking dependent risk of coronary disease, myocardial infarction and placental disruption. Similarly, variations in the VEGF gene have been associated with increased risk of various cancers, DiGeorge syndrome, psoriasis, diabetic renal disease and amyotropic lateral sclerosis. Understanding the molecular basis of these genetic variations and how contribute to the pathophysiology provides new and important insights into human disease.
Post-war Violence in Italy: A Struggle for Memory
Authors: Storchi Massimo
Published: July 2007

The Resistance at the root of Italian democracy is still the object of political and often controversial debate, 60 years on. This is related to the various phases of political development in Italy: from the early post-war years, characterized by the conflict associated with the ideological clashes of the Cold War, to the 1960s and 1970s, afflicted by terrorism, to the fall of the Berlin Wall and then to the collapse of the party system. Diverse, sometimes conflicting memories of the Resistance have emerged, linked not only to the numerous forms characterizing the struggle against Nazi–Fascism, but also to the varying motives, ideals and politics which animated fighters on both sides. With the new bipolar political system and the rise of the Right, the Resistance has returned to being one of the most prominent features of political controversy. This manifests itself in editorial strategies and extensive media operations in which memories representing those people who are opposed to the ideals of the Resistance seem to have the upper hand.
Characterisation of lipid‐protein interactions using a surface plasmon resonance biosensor
Authors: Rajaram O. V. , Sawyer W. H.
Published: July 2007

The binding of proteins to lipid surfaces is of fundamental importance when considering the mechanisms of lipases and lipid transfer proteins. Current procedures for determining the binding constants suffer from problems related to poor sensitivity, poor reproducibility and to the use of restrictive assumptions in the analysis. We demonstrate here the use of the surface plasmon resonance biosensor to determine the association and dissociation kinetics of binding of cholesteryl ester transfer protein to the surface of emulsion particles of defined size and composition. Uncertainty about the emulsion valency, that is, the number of sites on the lipid particle occupied by the immobilised transfer protein on the biosensor surface, means that the association kinetics can only be characterised in terms of the product of the association rate constant and the concentration of sites, whereas the dissociation rate constant can be obtained unambiguously. The technique will prove useful in characterising the binding of proteins to lipid surfaces.
Interactions of trans‐acting factors(s) with the estradiol response element and nuclear factor 1 of the vitellogenin II gene of Japanese quail
Authors: Gupta Sanjay , Upadhayay Rashmi , Kanungo M. S.
Published: July 2007

This study was directed at achieving an understanding of the mechanisms by which steroid hormones control the synthesis of vitellogenin (VTG) protein in the liver of the Japanese quail. Northern hybridization shows that administration of estradiol alone or with progesterone stimulates the synthesis of VTG mRNA. Gel mobility shift assay of DNA fragments containing the ERE and NF1 shows that estradiol alone or with progesterone increases the levels of nuclear proteins that bind to these cis‐acting elements of the promoter of VTG gene. The cooperative effect of the two hormones seen at the level of expression of the VTG gene may be due to protein‐protein interactions of trans‐acting factors that bind to ERE and NF 1.
Oxidative damages by iron‐chelate complexes depend on the interaction with the target molecules
Authors: Asaumi Akira , Ogino Tetsuya , Akiyama Takashi , Kawabata Teruyuki , Okada Shigeru
Published: July 2007

To elucidate that in iron‐catalyzed oxidative damage the interaction of iron complex with the target molecules is important, the oxidative damage to plasmid DNA, protein and fatty acid has been compared using iron‐chelate complexes with nitrilotriacetic acid (nta), citric acid, ethylenediamine‐N,N′‐diacetic acid (edda) and diethylenetriamine‐N,N,N′,N″,N″‐pentaacetic acid (dtpa). In the presence of hydrogen peroxide, plasmid pBR322 strand breaks occurred in the order of Fe‐edda > Fe‐citrate > Fe‐nta >> Fe‐dtpa. However, fragmentation of bovine serum albumin and diene conjugation of linoleic acid micelle occurred in the order of Fe‐nta > Fe‐edda >> Fe‐citrate > Fe‐dtpa = 0, which were similar to hydroxyl radical production by these iron complexes and H2O2. Bleomycin‐detectable free radical‐promoting irons in these iron complexes were about 85% of iron in Fe‐nta, Fe‐citrate and Fe‐edda, and only about 33% in Fe‐dtpa. Not only hydroxyl radical productivity and free radical‐promoting iron content in iron complex, but also the interaction of the complex with the target molecules determines the iron‐catalyzed oxidative damage.
Estrogen surge: A regulatory switch for superoxide radical generation at implantation
Authors: Laloraya Malini , Jain Sudhir , Thomas Monzy , Kopergaonkar Swati , Pradeep Kumar G.
Published: July 2007

This study is undertaken to identify the regulatory switch for the activation of superoxide radical generation pathway and repression of superoxide dismutase (SOD) at the time of implantation using estrogen as a control factor and delayed implantation as a model system. The results revealed high SOD activity and negligible oxyradical generation in progesterone‐treated animals on day‐5 and day‐8 (delayed implantation) while an enormous rise in oxyradical generation and an abrupt fall in SOD in animals which received both estrogen and progesterone injections were observed on days 5 and 8 of pregnancy. These results strongly suggest that estrogen regulates superoxide anion radical generation by lowering the SOD activity.
Role of the reductant substrates on the inactivation of horseradish peroxidase by m‐Chloroperoxybenzoic acid
Authors: Arnao M. B. , Garcia‐Cánovas F. , Acosta M.
Published: July 2007

Horseradish peroxidase reacts with H2O2 and other hydroperoxides to form Compound I, the first active enzymatic form. m‐Chloroperoxybenzoic acid, a xenobiotic hydroperoxide, acts as an oxidant substrate of horseradish peroxidase. However, this hydroperoxide is also a powerful inactivator of the enzyme and in this sense is more effective than H2O2. The coupled reductant substrates used in the peroxidatic reaction protect the enzyme from the inactivating process.

A reaction mechanism is proposed with two competitive routes: one catalytic and one inactivating. Using a kinetic approach, the ratio between the hydroperoxide and the reductant substrate appears to be a decisive factor in the catalytic turnover of the enzyme.

The role of the reductant substrates in protecting the enzyme, and the physiological and biotechnological implications of this process are discussed.
Regulation of the activity of M‐phase promoting factor through protein kinase A‐mediated pathway in LP1‐1 cells
Authors: Oh Won Jong , Joe Cheol O. , Choi Kyung Hee
Published: July 2007

Treatment of cAMP analogs, dibutyryl cAMP and 8‐bromo‐cAMP, was found to inhibit the proliferation of mouse fibroblast LP1‐1 cells and the p34cdc2 kinase activity of M‐phase promoting factor (MPF). However, it showed relatively little effect on expression of the cyclin B1 and cdc2 genes. On the other hand, when the nuclear extracts obtained from the cells at early G2 phase were treated with cAMP analogs, the kinase activity was significantly decreased as compared to the untreated control. Furthermore, the inhibitory effect of cAMP analogs could be reversed upon treating with okadaic acid even in the presence of the cAMP analogs, implying that cdc25 remains in an active form. In addition, the treatment of okadaic acid stimulated the cell progression. These results suggest that down‐regulation of MPF activity through protein kinase A‐mediated pathway is under post‐translational control and cdc25 activation pathway involving okadaic acid‐sensitive phosphatase play a role in the regulation of MPF activity.
Zearalenone ‐ Induced uncoupling in plant mitochondria is sensitive to 6‐ketocholestanol
Authors: Macri F. , Vianello A. , Braidot E. , Petrussa E. , Mokhova E. N.
Published: July 2007

Zearalenone (F‐2) is a mycotoxin which acts as a protonophoric uncoupler in plant mitochondria [Macri, F. and Vianello, A., (1990) J. Plant Physiol. 136, 754‐757]. In the present paper, the mechanism of F‐2‐induced uncoupling in pea mitochondria was studied. The uncoupling by F‐2 was partially reversed by 6‐ketocholestanol (kCh) under conditions in which kCh completely reversed the FCCP‐induced uncoupling and almost did not affect the palmitate‐induced uncoupling. Recoupling effects of carboxyatractylate, ADP and cyclosporin A were small and could not essentially decrease the kCh‐insensitive part of F‐2‐induced uncoupling. It is suggested that a protein, mediating kCh‐sensitive uncoupling, is involved in the F‐2 effect in plant mitochondria.
Interaction of myosin subfragment 1 with F‐actin studied by differential scanning calorimetry
Authors: Nikolaeva Olga P. , Orlov Victor N. , Dedova Irina V. , Drachev Vladimir A. , Levitsky Dmitrii I.
Published: July 2007

The thermal unfolding of the myosin subfragment 1 (S1) and of filamentous actin (F‐actin) in their strong complex obtained in the presence of ADP was studied by differential scanning calorimetry (DSC). It is shown that in the acto‐S1 complexes S1 and F‐actin melt separately, and thermal transitions of each protein can be easily followed. Interaction of S1 with F‐actin significantly increases S1 thermal stability and also affects the thermal stability of F‐actin. Although S1 unfolds at much lower temperature than F‐actin, the molecules of S1 remain bound to F‐actin even after full denaturation. Under these conditions S1 may induce cross‐linking between actin filaments. It is concluded that DSC studies on the acto‐S1 complexes offer a new and promising approach to investigate the structural changes which occur in the myosin head and in F‐actin due to their interaction.
Mapping the 90 kDa heat shock protein binding region of the Ah receptor
Authors: Perdew Gary H. , Bradfield Christopher A.
Published: July 2007

Expression of a series of Ah receptor (AhR) deletion mutants in an in vitro translation system has been previously used to map several functional domains of the murine AhR (Dolwick et al. (1993) Proc. Natl. Acad. Sci. USA 90, 8566‐8570). In this report, quantitative immunoprecipitation of 90‐kDa heat shock protein (hsp90) from reticulocyte lysate allowed us to measure the level of the AhR and AhR deletion mutants complexed with hsp90. After translation of a series of deletion mutants it was determined that there are two distinct domains important in forming a stable AhR/Hsp90 complex, corresponding to amino acid sequences 1‐166 and 289‐347 of the AhR. Neither ARNT, nor Per were able to stably interact with hsp90. Thus, the AhR appears to be a unique member of the PAS domain family of proteins that binds a known ligand and stably interacts with hsp90.
Molecular cloning and expression of a cDNA encoding a protein tyrosine phosphatase from mouse brain
Authors: Park Kyung Soo , Kim Yeon Woong , Kim Choong Won , Kang Dae Ook , Byun Si Myung
Published: July 2007

A mouse brain cDNA library was screened with a radioisotope‐labeled probe of human placental protein tyrosine phosphatase. The isolated clone (MBPTP1B) was found to contain an open reading frame of 1,296 nucleotides as well as 5′ (709 nucleotides) and 3′ (341 nucleotides) non‐coding regions. This cDNA encodes a PTP of 432 amino acids having a mass of 49,563 daltons and exhibiting 83% and 93.5% sequence identity to that of human PTP1B and rat PTP1, respectively. The expression of the cDNA in yeast was identified by western blot analysis and PTP activity assay.
Hyaluronic acid induced hyaluronic acid binding protein phosphorylation and inositol triphosphate formation in lymphocytes
Authors: Rao Ch. Mastan , Deb Tushar Baran , Datta Kasturi
Published: July 2007

In this report, the role of 34 kDa HA‐binding protein in hyaluronic acid‐induced cellular signalling in lymphocytes has been examined. The binding of 125I‐HA to lymphocytes in vivo was found to be inhibited by pre‐incubation of the cells with anti‐34 kDa HA‐binding protein antibodies, thus confirming 34 kDa HA‐binding protein as the specific HA‐receptor in lymphocytes. This observation was substantiated by anti‐34 kDa HA‐binding protein antibodies immunoblotting and 125I‐HA ligand blotting of lymphocytes cell lysate. The HA‐induced cell aggregation, tyrosine phosphorylation and cytoskeletal protein phosphorylation demonstrate the HA‐induced early cellular signalling events in lymphocytes. Further, to study the involvement of 34 kDa HA‐binding protein in mitogen induced lymphocyte signalling, we studied in vivo phosphorylation and secondary messenger formation. The enhanced 34 kDa HA‐binding protein phosphorylation by HA and the inhibition of cellular aggregation and IP3 formation by anti‐HA‐binding protein antibodies revealed that 34 kDa HA‐binding protein is one of the potential mediators in HA‐induced signal transduction.
Development of a metallothionein based heavy metal biosorbent
Authors: Pazirandeh Mehran
Published: July 2007

The potential utility of a recombinant E. coli expressing the Neurospora crassa metallothionein gene (NCP) as a heavy metal biosorbent was investigated. It was shown that the NCP was capable of efficiently removing low levels of several metals (including cadmium, lead, and mercury) from solutions. The reusability of the NCP was demonstrated through 5 cycles of metal binding, stripping with dilute acid, and regeneration of the binding sites with out any adverse effect on the metal binding activity. The NCP was successfully encapsulated in alginate and acrylamide without any inhibitory effect on its metal uptake activity. Furthermore, the metal uptake activity of the NCP was shown to be metabolism independent and resistant to solvents and other compounds (eg. polyaromatic hydrocarbons) which are often present along with heavy metals in waste waters thereby creating the potential for non‐viable, encapsulated cells to be used.
The fungus Gibberella fujikuroi produces copper/topaquinone‐containing amine oxidase when induced by n‐butylamine
Authors: Frébort Ivo , Matsushita Kazunobu , Adachi Osao
Published: July 2007

Crude extract of Gibberella fujikuroi AKU 3802 mycelium induced with n‐butylamine showed a single amine oxidase activity band in a non‐denaturing gel that cross‐reacted with the antibody against copper/topaquinone‐containing amine oxidase from Aspergillus niger. The enzyme was purified by a procedure involving four chromatographic steps. Purified enzyme was pink with an absorption maximum at 490 nm. Molecular mass of 135 kDa estimated by gel chromatography and 70 kDa found by SDS‐PAGE confirmed the dimeric structure of the enzyme. The enzyme readily oxidized n‐hexylamine, n‐butylamine, benzylamine and histamine, but not spermine or spermidine. Inactivation by carbonyl reagents and copper chelators suggested the presence of a copper/topaquinone cofactor. Spectrophotometric titration by p‐nitrophenylhydrazine showed one reactive carbonyl group per subunit and redox‐cyclic quinone staining confirmed the presence of a quinone cofactor, pH‐dependent shift of the absorption spectrum of the enzyme p‐nitrophenylhydrazone (465 nm at neutral to 580 nm at alkaline pH) supports the identity of the cofactor with topaquinone. The N‐terminal amino acid sequence of the enzyme showed high similarity to other microbial copper/topaquinone‐containing amine oxidases.
Effect of polyanions on ferricytochrome c
Authors: Sedlák Erik
Published: July 2007

Effects of two polyanions (heparin, Nation) on absorption and CD spectra in the Soret region, and on fluorescence of ferricytochrome c were investigated. The conformational state of ferricyt c in the complexes with studied polyanions is significantly different. Whereas heparin affects the structure in the heme region of ferricyt c only slightly (ferricyt c remains in a native‐like state), the influence of Nation is more expressive (the Met‐80 ‐ heine iron bond and the deeply buried region around Trp‐59 are perturbed). Shift of the pK of the transition from neutral low‐spin state to acidic high‐spin state to higher pH values was observed.
Separation of three P450 isozymes from liver microsomes of gilthead seabream treated with ß‐NF and partial purification of cytochrome P4501A1
Authors: Sen Alaattin , Arıç Emel
Published: July 2007

Three distinct cytochrome P450 isozymes, P4501A1, P4502B and an unidentified P450 isozyme were separated and isolated from ß‐NF‐treated gilthead seabream liver microsomes. Cytochrome P4501A1 was partially purified from ß‐NF‐treated gilthead seabream liver microsomes in the presence of detergents Emulgen 913 and cholate and protease inhibitors using two DEAE‐cellulose, Porapak Q and two hydroxylapatite column chromatographies. The overall yield of purified P4501A1 was 1.2% with respect to microsomal total P450 with a specific content of 3 nmol/mg protein. The purified P4501A1 was characterized with respect to spectral, electrophoretic, biocatalytic and immunochemical properties, which are found to be similar to P4501Als purified from other teleost species such as trout, scup, cod and perch suggesting that the P450 we have purified belongs to CYP1A1.
Free radical‐induced inhibition of ATP diphosphohydrolase activity (EC 3.6.1.5) from rat blood platelets
Authors: Frassetto Silvana S. , Dias Renato D. , Sarkis João J. F.
Published: July 2007

In the present report we demonstrate the in vitro effects of free radicals on an ecto‐ATP diphosphohydrolase activity (apyrase, EC 3.6.1.5) from rat blood platelets. Rat blood platelets were exposed to an oxidant‐generating system (H202 / Fe2+ / ascorbate) and the ATP diphosphohydrolase activity was inhibited. The enzyme inhibition was prevented by glutathione (GSH) and cysteine but not by trolox as a vitamin E analogue. The TBARS (thiobarbituric acid reactive substances) assay and the determination of sulphydryl groups indicate that the inhibition of the enzyme activity in resting platelets is not related to lipid peroxidation or to oxidation of sulphydryl residues. These results demonstrate the susceptibility of ATP diphosphohydrotase activity from platelets to free radicals and suggest that amino acid residues which are essential for the enzyme function are probably modified.
Effect on product specificity of cyclodextrin glycosyltransferase by site‐directed mutagenesis
Authors: Kim Young Ho , Bae Kwang Hee , Kim Tae Jip , Park Kwan Hwa , Lee Hyun Soo , Byun Si Myung
Published: July 2007

Cyclodextrin glycosyltransferase (CGTase; EC 2.4.1.19) catalyzes the degradation of starch into cyclodextrins through an intrarnolecular transglycosylation reaction. Tyr‐89, Asn‐94, and Tyr‐100 are located near the putative active center. To analyze their roles in product specificity, Tyr‐89, Asn‐94, and Tyr‐100 of CGTase from alkalophilic Bacillus sp. 1‐5 were replaced with different amino acids. Among the mutants, the N94S mutant protein produced about two times more α‐cyclodextrin than the wild‐type at all incubation times. The Y89F and Y100F mutant proteins were changed to more β‐specific enzymes. From these results it is suggested that the changing of the residues located at the near active site can change the product specificity of CGTase.
Molecular cloning and nucleotide sequencing of bioF (7‐keto‐8‐amino pelargonic acid synthetase), BioC and BioD (dethiobiotin synthetase) genes of Erwinia herbicola
Authors: Wu Chwen‐Huey , Bao Yu‐Yen , Shao Chung‐Ping , Shiuan David
Published: July 2007

The biotin operon of Erwinia herbicola Eho 10 was cloned and characterized by complementation of E. coli biotin mutants. The operon was found to contain five genes arranged in the order, bioABFCD. The nucleotide sequences of bioF (7‐keto‐8‐aminopelargonic acid synthetase), bioC and bioD (dethiobiotin synthetase) were determined and analyzed. The nucleotide sequences and deduced amino acid sequences of bioFCD were compared with the corresponding sequences from Escherichia coli, Bacillus sphaericus, Serratia marcescens and Brevibacterium flavum.
Application of hammerhead ribozymes for structural studies of ribosomal 5S RNAs
Authors: Szymañski Maciej , Fürste Jens‐Peter , Barciszewska Miroslawa Z. , Erdmann Volker A. , Barciszewski Jan
Published: July 2007

We synthesized and used hammerhead ribozymes designed for the GUC and AUC target sequences located in the terminal loop C of animal and plant 5S rRNAs to verify its involvement in a higher order structure formation of ribosomal RNA. The activity of both GUC and AUC specific ribozymes were proven to be active against synthetic analogs of the stem III and loop C in the two types of 5S rRNAs. In contrast we did not observe any ribozyme‐catalysed hydrolysis within the loop C of the native 5S rRNAs. We considered two possible explanations of the results: an involvement of the loop C in long range interactions and lack of a structural flexibility to form proper tertiary structure of the complex with the ribozyme. From our data it seems that the single stranded nucleotides within the loop C can not form correct RNA duplex structure required for ribozyme activity. It is clear that hammerhead ribozymes can be used for inhibition of different steps of expression of genetic information only if the target sequence occurs in appropriate context, that allows to form correct secondary and tertiary structure of the ribozyme‐substrate complex.
Metabolic fate and effects of saturated and unsaturated fatty acids in Hep2 human larynx tumor cells
Authors: Colquhoun Alison , Curi Rui
Published: July 2007

Previous studies have reported the presence of carnitine palmitoyltransferase I and II in tumor cells and the inhibitory effects of fatty acids on cell proliferation. The present work considered the metabolic fate of [14C] or [3HI‐labeled fatty acids and their effects on cellular metabolism in Hep2 human larynx tumor cells.The rate of uptake of acetate was 45% of that of myristate, palmitate, oleate, linoleate and arachidonate. However, acetate was rapidly metabolized within the cell as seen by its low rate of accumulation as non‐esterified fatty acid, <5% of that of the other fatty acids. The incorporation of fatty acids into neutral lipid fractions showed palmitate and oleate primarily entered the phospholipid fraction, while linoleate and arachidonate entered equally the phospholipid and triacylglycerol fractions. Palmitate and oleate were oxidized to 14CO2 at higher rates than linoleate and arachidonate, with arachidonate being the least oxidized of the unsaturated fatty acids. Acetate was oxidized at 10‐30 fold higher rates than the other fatty acids. Palmitate, oleate, linoleate and arachidonate all had significant inhibitory effects on the rate of glucose utilization by Hep2 cells, ranging from 25‐38% inhibition and were found to inhibit cell proliferation by 17‐73%. These findings suggest that certain fatty acids not only play a structural role in cellular metabolism, but may also have a potential regulatory role in the glycolytic pathway of Hep2 cells.
Enhanced production of LTB4 and free radicals in rat lung by exhaustive physical exercise
Authors: Kumar T. Charles , Reddy K. Veera , Anuradha D. , Reddanna P.
Published: July 2007

Leukotrienes formed by the action of 5‐lipoxygenase on arachidonic acid are a group of endogenous compounds active in hypersensitivity reactions and inflammation. In this report the formation of LTB4 from LTA4 was measured by incubating lung microsomes from normal and exercised rats to determine whether LTB4 may have a role in the mediation of exercise‐induced oxidant stress. Exhaustive exercise in the form of swimming results in the enhanced production of LTB4 in lung microsomes. Further, free radicals measured by ESR spectroscopy were also significantly increased in lung tissue from exercised rats. These results suggest a role for LTB4 in the mediation of lung tissue damage during exercise‐induced oxidative stress.
The protective effects of dihydrolipoamide and glutathione against photodynamic damage by Al‐phtalocyanine tetrasulfonate
Authors: Kliukiené Regina , Maroziene Audroné , Nivinskas Henrikas , Čénas Narimantas , Kirveliené Vida , Juodka Benediktas
Published: July 2007

In spite of well‐known ability of lipoamide/dihydrolipoamide (LipS2NH2/Lip(SH)2 NH2) and oxidized/reduced glutathione (GSSg/GSH) couples to scavenge singlet oxygen (1O2), the possible protective effects of these compounds against photodynamical damage by Al‐phtalocyanine tetrasulfonate (Al‐PcS4) were examined. Using erythrocyte glutathione reductase, pig heart lipoamide dehydrogenase and hamster kidney fibroblast culture as model systems, we have found that protective effects of Lip(SH)2NH2 and LipS2NH2 were close to that of azide, far exceeding the effects of GSH and GSSG, and paralleling the rates of Al‐PcS4‐sensitized photooxidation of these compounds. We have failed to observe a previously described (Devasagayam, T.P.A., et al. (1991) Biochim. Biophys. Acta 1088, 409‐412) enhancement of damaging action of 1O2 by GSH. These findings point out to the possibility of LipS2NH2/Lip(SH)2NH2 to neutralize the side‐effects of photodynamic therapy, and to a minor but definitely protective role of GSH.
Renaturation and purification of recombinant tissue‐type plasminogen activator expressed in E. coli
Authors: Hua Zi‐Chun
Published: July 2007

Under the control of trp promoter, human tissue‐type plasminogen activator was expressed in E. coil in the form of inclusion body. The recombinant t‐PA was recovered for renaturation from preparative native PAGE gel by zinc acetate staining and electroelution. After renaturation in vitro, the recombinant t‐PA was purified by benzamidine affinity chromatography and lysine affinity chromatography. The purified t‐PA showed homogeneous on silver‐stained SDS‐PAGE gel, with a specific activity of 240,000I.U./mg protein.
An essential tryptophan residue of green crab (Syclla Serrata) alkaline phosphatase
Authors: Zheng Wen‐Zhu , Chen Qing‐Xi , Zhao Hong , Zhang Zhe , Zhang Wei , Zhou Hai‐Meng
Published: July 2007

The tryptothan residues in green crab (scylla serrata) alkaline phosphatase (EC 3.1.3.1) have been modified by N‐bromosuccinimide (NBS). The modification of five tryptophan residues leads to complete loss of enzymatic activity. With the increase of NBS concentration, both the absorption at 278 nm and the fluorescence emission intensity at 335 nm of the modified enzyme decreased markedly indicating the modification of tryptophan residues. Quantitative treatment of the data (Tsou, Sci. Sinica 1962, 11, 1535‐1558) shows that among the tryptophan residues modified, one is essential for its catalytic activity. The presence of the substrate markedly protects the modification of tryptophan residues as well as the inactivation, suggesting that the essential tryptophan residue is situated at the active site of this enzyme.
The large ribosomal protein gene cluster of a cryptomonad plastid: Gene organization, sequence and evolutionary implications
Authors: Wang Sheng‐Long , Liu Xiang‐Qin , Douglas Susan E.
Published: July 2007

The complete sequence of the major ribosomal protein gene cluster of the plastid genome of the cryptomonad alga Guillardia theta (formerly Cryptomonas Φ) is presented. The ribosomal protein genes (corresponding to the SIO, spc, alpha and L13/S9 operons of E. coli are found upstream of the previously reported plastid str operon, and transcribed in the same orientation. The genes are very tightly packed with as little as two nucleotides between the rpll4 and rpl24 genes. The gene arrangement is very similar to that reported for the rhodophyte alga, Porphyra purpurea, and the chromophyte diatom, Odontella sinensis, indicating a close evolutionary relationship between these groups of algae. Northern analysis indicates that the 29 genes1 are arranged as one operon and are transcribed as a single mRNA that is subsequently processed into smaller transcripts.
High‐level expression of cecropin CMIV in E. coli from a fusion construct containing the human tumor necrosis factor
Authors: Wang Liang , Wu Haihong , Dou Fei , Xie Wei , Xu Xianxiu
Published: July 2007

Cecropin CMIV gene was fused to the 3′‐terminus of the mutated tumor necrosis factor (TNFb) gene and the fusion gene was directly under the control of an inducible T7 promoter in pET‐11d. This fusion gene was overexpressed in Escherichia coli with an expression level of approximate 40%‐50% of total cellular proteins, and was produced mainly in the form of inclusion body. Peptide with antibacterial activity was obtained by cleaving the fusion protein with CNBr.
Evidence for antiradical and antioxidant properties of four biologically active N,N‐Diethylaminoethyl ethers of flavaone oximes: A comparison with natural polyphenolic flavonoid rutin action
Authors: Metodiewa Diana , Kochman Agata , Karolczak Stefan
Published: July 2007

The antiradical and antioxidant activities of four biologically active N,N‐diethyloaminoethyl ethers of flavanone oximes (N,N‐DEAEFo) were investigated in vitro and compared with these of polyphenolic flavonoid (rutin). Four experimental models were used: iron‐ and ascorbate driven Fenton systems, γ‐radiolysis, xanthine/xanthine oxidase system and diphenylpicrylhydrazyl (DPPH) radical scavenging. The results clearly indicate that N,N‐DEAEFo are acting as promising antioxidants and radioprotectors comparable to rutin activities: they should prove to be useful under acute oxidative stress conditions for which both properties were required.
Substances in the aqueous fraction of cigarette smoke inhibit lipid peroxidation in synaptosomes of rat cerebral cortex
Authors: Kamisaki Yoshinori , Wada Kouichirou , Nakamoto Kentaro , Kishimoto Yosuke , Ashida Kumiyo , Itoh Tadao
Published: July 2007

The effects of water‐soluble substances in cigarette smoke on lipid peroxidation were investigated using nerve terminals prepared from the rat cerebral cortex. The prepared smoke‐substances significantly reduced the spontaneous increase in thiobarbituric acid‐reactive substances in synaptosomes in a dilution factor‐dependent manner. Furthermore, the aqueous extract also inhibited the elevation of lipid peroxidation induced by 2,2′‐azobis (2‐amidinopropane) dihydrochloride, a peroxyl radical generator. Smoke‐substances scavenged superoxide radicals generated from stimulated human leukocytes and from the xanthine/xanthine oxidase system. These effects were not mimicked by nicotine. The antioxidant effects of smoke‐substances were preserved for several days at 5 ° or ‐80 °C. The results suggest that the smoke‐substances may possess long half‐lives and scavenge the radicals which cause lipid‐peroxidation in synaptosome membranes.
Ontogeny of neurotransmitter amino acids in human fetal brains
Authors: Das Shyamal K. , Ray Pradip K.
Published: July 2007

A wide spectrum of developmental profiles is presented for a few important neurotransmitter amino acids, namely γ‐aminobutyric acid, glycine, glutamic acid, aspartic acid and taurine as well as the key enzymes involved in their metabolism in human fetal brains of different gestational ages. Besides taurine almost all these amino acids and their key metabolic enzymes were found to be progressively enhanced upto the mid period of third trimester of pregnancy indicating a rapid growth of nerve processes, myelination and maturation of fetal brains particularly during this period. Interestingly taurine showed an inverse relationship of ontogenic pattern with respect to its biosynthesizing enzyme in fetal brains.
Glucose transport in a murine mammary epithelial cell line
Authors: Bennett Brydon L. , Grigor Murray R. , Prosser Colin G.
Published: July 2007

The glucose transport systems of the COMMA‐D cell line (a murine mammary epithelial cell line) were examined using 2‐deoxyglucose as substrate. The kinetics and inhibition studies with other sugars including xylose suggested that the transport system had properties of both GLUT‐1 and Gtut‐3. Subsequent analysis of mRNA transcripts using cDNAs for GLUT‐1 to 4 showed that only GLUT‐1 was expressed in the COMMA‐D cells. The results highlight the fact that kinetic and substrate specificity are not sufficient, by themselves, for the identification and characterisation of GLUT isoforms in cultured cells.
Partial restoration of inactivated ribosomes: role of the aldehyde group generated by RNA N‐glycosidase in the sarcin/ricin domain of 28S rRNA in ribosome
Authors: Cheng Le‐Ping , Liu Wang‐Yi
Published: July 2007

In this paper, we report more information on the important role of the aldehyde group in the sarcin/ricin domain of 28S rRNA in rat liver ribosome. We find D‐amino acids, amino acid derivatives having free amino group and two polyamines can also partially restore the activity of cinnamomin‐inactivated ribosomes. However, amino acid derivatives and a tripeptide with blocked amino group cannot. Neither sodium borohydride nor the L‐amino acids can restore the activity of ribosomes inactivated by α‐sarcin. These data demonstrate that partial restoration of the activity of the inactivated ribosome is indeed the result of the blockage of the aldehyde group. It reaches the conclusion that emergence of the active aldehyde group in the sarcin/ricin domain of 28S rRNA is one of factors that inactivate the ribosome for protein synthesis.
Production and characterisation of monoclonal and polyclonal amtobpdoes agaomst bovine osteocalcin
Authors: Nielsen Peter Kresten , Hønberg Lisbeth Skou , Larsen Torben , Sperling‐Petersena Hans Uffe , Sottrup‐Jensen Lars
Published: July 2007

Monoclonal and polyclonal antibodies were produced against osteocalcin purified from bovine bone. The antibodies were used for the development of a two‐site ELISA. A monoclonal antibody recognising residues 15‐49 of bovine osteocatcin generated by cleavage with 70% formic acid was used as capture antibody and a biotinylated polyclonal antibody recognising the same peptide was used for detection. The two‐site assay did not detect any tryptic peptides of bovine osteocalcin (residues 1‐19, 20‐43, and 45‐49) nor did it detect the peptide 1‐14 or the synthetic peptide 37‐49. Cross‐reaction was observed for osteocalcin in plasma from sheep, goat, chicken and human but not for osteocalcin from rat and pig.
Synergistic effect of glycolic acid on the antioxidant activity of α‐tocopherol and melatonin in lipid bilayers and in human skin homogenates
Authors: Morreale Massimiliano , Livrea Maria A.
Published: July 2007

Considerable interest has been raised concerning the use of natural compounds in preventing skin aging and photoaging. In the idea that the combined action of agents increasing epidermal turnover with antioxidants could be advantageous in cosmetic and therapeuthic treatments, we first investigated if α‐glycolic acid affected or prevented the antioxidant activity of vitamin E and of melatonin, two compounds found beneficial as topical photoprotectant. Assays were carried out in vitro either in a biomimetic liposomal system, or in human skin homogenates. Lipid peroxidation was monitored spectrophotometrically by the time course of lipid hydroperoxide production in liposomes and by formation of TBA reactive substances (TBARS) in skin homogenates. Glycolic acid. at 25 μM to 1 mM, showed a mild, concentration‐dependent antioxidant effect in liposomes, as evaluated by a slight decrease of the peroxidation rate, while, at 1 mM, reduced TBARS production in skin homogenates by 14 %. Combinations of either vitamin E or melatonin with glycolic acid, in a 1:5 to 1:200 molar ratio, resulted in a clear synergistic protection of liposomes, more evident for the combination of glycolic acid with vitamin E. An amount of synergism up to 250% and up to 80% was evahmted with vitamin E and melatonin, respectively. Consumption rate of vitamin E during peroxidation of liposomes, in the absence or in the presence of glycolic acid, suggests that regeneration of vitamin E may in part explain the observed synergism. Synergistic antioxidant activity between vitamin E and glycolic acid was also observed in skin homogenates, whereas the effect of glycolic acid on the antioxidant activity of melatonin appeared additive. However, the combination of these three compounds inhibited TBARS production almost completely.

Our data provide evidence that glycolic acid can strongly potentiate the antioxidant action of melatonin and vitamin E. This may suggest the advantage of combining α‐glycolic acid with these antioxidants in skin designed preparations, both to improve penetration and availability of antioxidants to epidermal layers and to enhance their protective potential.
Evidence that Fe‐NTA‐induced renal prostaglandin F2α is responsible for hyperplastic response in kidney: Implications for the role of cyclooxygenase‐dependent arachidonic acid metabolism in renal tumor promotion
Authors: Iqbal Mohammad , Giri Uma , Giri Dipak K. , Athar Mohammad
Published: July 2007

Oxidative stress in a tissue activates phospholipase A2 which releases free arachidonic acid. In addition, a low grade oxidative tone also stimulates the tissue cyclooxygenase activity. Cyclooxygenase‐dependent arachidonic acid metabolites such as PGF2α are known to play an important role in the development and maintenance of hyperplasia in skin in response to the application of tumor promoters. In this study we show that Fe‐NTA, an oxidant renal tumor promoter induces PGF2α which was maximum at 12 hours after Fe‐NTA treatment. However, at all time points studied, the elevated levels of PGF2α have been observed. As a result of the induction of PGF2α, the hyperplastic response can also be observed in the histopathology of the tissue. Additionally, an increased incorporation of [3H]thymidine in renal DNA has also been observed. Pretreatment of animals with indomethacin suppresses Fe‐NTA‐mediated hyperproliferation suggesting a role of cyclooxygenase in Fe‐NTA‐mediated stimulation of hyperplastic activity. The pretreatment of animals with the chain breaking antioxidants, Vit. E, BHA and BHT were only partially effective in inhibiting Fe‐NTA‐mediated PGF2α production, further suggesting a role of non‐free radical‐dependent mechanism in its production. Our data suggest that Fe‐NTA‐induced PGF2α through the activation of cyclooxygenase is responsible for the development and maintenance of hyperplasia in kidney.
The increased level of PDGF‐A contributes to the increased proliferation induced by mechanical stimulation in osteoblastic cells
Authors: Wang Wei , Zhuang Hongming , Levitz Craig L. , Fan Huijun , Seldes Richard M. , Tahernia A. David , Brighton Carl T.
Published: July 2007

Mechanical stimulation can prompt healing of bone fractures. However, it is largely unknown how osteogenesis is promoted by mechanical stimulation. In this study, we found that mechanical strain‐induced proliferation of osteoblastic cells (MC3T3‐E1) accompanied increased levels of platelet‐derived growth factor‐A (PDGF‐A) mRNA, determined by quantitative reverse transcription/polymerase chain reaction. In addition, neomycin and W‐7, which blocked mechanical strain‐induced proliferation of the osteoblast cells, also blocked mechanical stimulation‐induced elevation of PDGF‐A mRNA. Finally, an antibody against PDGF can inhibit physical stimulation‐induced proliferation of MC3T3‐E1 ceils, suggesting that the increased MC3T3‐E1 cells produced by mechanical stimulation at least partially depends on the increased activity of PDGF.
Studies on growth‐promoting action of insulin: mitogenic activity of insulin and its analogues in mouse mammary tumor cells
Authors: Shi Min , Feng You‐Min
Published: July 2007

Growth‐promoting action of insulin and its analogues was studied in the mouse mammary tumor cell line GR2H6 by measuring 3H‐thymidine incorporation. The mitogenic activity of the analogues obtained by shortening of C‐terminus of insulin B chain decreased with stepwise removal of terminal residues, and [A3Leu]insulin was approximately 1000 times less potent than insulin. However, the despentapeptide(B26‐B30) insulin(DPI) showed a strikingly higher mitogenic activity at high concentrations; the detected maximal effect of DPI was 136% more than that of insulin. These results demonstrate that the C‐terminal area of the B chain and the N‐terminal α‐helix of the A chain are important for the mitogenic activity of insulin. A region corresponding to the growth‐promoting function of insulin and a model of DPI action are proposed and discussed.