bluuebunny/crossref_metadata_2025 huggingface dataset:
Role for c-Jun N-Terminal Kinase in β-Cell Recovery from Nitric Oxide-Mediated Damage
Authors: Scarim Anna L. , Nishimoto Sheri Y. , Weber Sarah M. , Corbett John A.
Published: May 2003

Treatment of rat islets with the cytokine IL-1 results in the inhibition of mitochondrial function and insulin secretion, events that are mediated by β-cell expression of iNOS [inducible nitric oxide (NO) synthase] and production of NO. β-Cells recover from the inhibitory actions of NO, produced following 24 h incubation with IL-1, on islet oxidative metabolism and insulin secretion if iNOS enzymatic activity is inhibited and the islets are cultured (in the presence of IL-1 and iNOS inhibitors) for a brief period of 8 h. Islet recovery from cytokine- and NO-mediated damage is an active process that requires new gene expression, and NO itself is one activator of this recovery process. In this study, the mechanism by which NO stimulates islet recovery has been examined. Incubation of rat islets or RINm5F cells with the NO donor compound, sodium (Z)-1(N,N-diethylamino) diazen-1-ium-1,2-diolate (DEA-NO) for 1 h results in a 60% inhibition of mitochondrial aconitase activity. β-Cells completely recover aconitase activity if the cells are washed to remove the NO donor compound and incubated for an additional 5 h in the absence of DEA-NO. The recovery of mitochondrial aconitase activity correlates with a 4-fold increase in cyclic GMP accumulation and is prevented by the inhibition of guanylate cyclase. The recovery of aconitase activity also correlates with the activation of members of the MAPKs, p38, c-Jun N-terminal kinase (JNK) and ERK, and the activation p38 and JNK is attenuated by inhibition of guanylate cyclase. ERK and p38 do not appear to participate in the recovery process as selective inhibition of these kinases fails to prevent recovery of aconitase activity; however, transduction of β-cells with a dominant negative mutant JNK prevents β-cell recovery from NO-mediated damage. These findings support a role for guanylate cyclase and JNK in the recovery of β-cells from NO-mediated damage.
Corticosterone Enhances Adrenocorticotropin-Induced Calcium Signals in Bovine Adrenocortical Cells
Authors: Chiyo Tomoko , Yamazaki Takeshi , Aoshika Kenji , Kominami Shiro , Ohta Yoshihiro
Published: May 2003

The rapid effects of steroid hormones on Ca2+ signals have been examined in bovine adrenocortical cells. Among the steroid molecules tested, only corticosterone rapidly stimulated Ca2+ signals upon addition of ACTH, although corticosterone alone did not induce Ca2+ signals. Corticosterone also enhanced steroidogenesis induced by ACTH. The enhancement of ACTH-induced Ca2+ signals was also observed with membrane-impermeable corticosterone conjugated to BSA and was not inhibited by cycloheximide. In addition, corticosterone did not enhance Ca2+ signals induced by ATP or angiotensin II. These results suggest that corticosterone selectively stimulates ACTH-induced Ca2+ signals in a nongenomic way by acting on a target in the plasma membrane. Furthermore, the supernatants of cells incubated with ACTH or ATP enhanced Ca2+ signals, suggesting that steroids produced by such treatment act in an autocrine fashion. Consistent with this idea, these effects were inhibited by inhibitors of steroidogenesis (aminoglutethimide or metyrapone). These results show that steroid molecules synthesized in adrenocortical cells facilitate ACTH-induced Ca2+ signals. Taken together, corticosterone secreted from adrenocortical cells activates ACTH-induced Ca2+ signals and steroidogenesis by nongenomic means.
Palm representation and approximation of the covariance of random closed sets
Authors: Böhm Stephan , Schmidt Volker
Published: May 2003

The covariance C(r), r ≥ 0, of a stationary isotropic random closed set Ξ is typically complicated to evaluate. This is the reason that an exponential approximation formula for C(r) has been widely used in the literature, which matches C(0) and C(1)(0), and in many cases also limr→∞C(r). However, for 0 < r < ∞, the accuracy of this approximation is not very high in general. In the present paper, we derive representation formulae for the covariance C(r) and its derivative C(1)(r) using Palm calculus, where r ≥ 0 is arbitrary. As a consequence, an explicit expression is obtained for the second derivative C(2)(0). These results are then used to get a refined exponential approximation for C(r), which additionally matches the second derivative C(2)(0).
Technology of Offshore Wind Turbines and Farms and Novel Multilevel Converter-Based HVDC Systems for Their Grid Connections
Authors: Agelidis Vassilios G. , Mademlis Christos
Published: May 2003

The technology associated with offshore wind farms is discussed in detail. First, the various offshore wind turbines are reviewed and the factors influencing their characteristics are outlined in comparison with their onshore counterparts. This overview serves as a basis for the discussion that follows regarding the possible electrical connection within the farm, and between the farm and the grid. Voltage-source converter-based HV DC connection is compared with HVAC connection. Finally, a novel multilevel converter-based HVDC system, based on flying capacitor multilevel converters is proposed, as a possible interface between the farm and the grid.
Embedded Wind Turbine Generation into AC Weak Distribution Grids
Authors: Schinas N. , Vovos N. A. , Giannakopoulos G. B.
Published: May 2003

A specific wind turbine (WT) generation scheme is used and a control strategy is studied, not only to increase the efficiency of the WT but also to limit the influence on the alternating current (AC) distribution grid. The system is a variable-speed wind-energy conversion scheme, including a synchronous generator with two three-phase stator windings displaced by 30°, which are connected to the AC distribution grid through DC links with Voltage Source Inverters (VSI). A state feedback controller has been designed, using the pole placement technique, to maximize the wind-energy capture. The controller of the DC link secures a smooth supply of the WT electric power to the distribution AC grid by supporting the output voltage and reducing the harmonic distortion. The operation of the proposed system and the efficiency of the control strategy are demonstrated and tested using simulation techniques.
An investigation into alleged ‘hauntings’
Authors: Wiseman Richard , Watt Caroline , Stevens Paul , Greening Emma , O'Keeffe Ciarán
Published: May 2003

In cases of alleged hauntings, a large number of seemingly trustworthy witnesses consistently report experiencing unusual phenomena (e.g. apparitions, sudden changes in temperature, a strong sense of presence) in certain locations. The two studies reported here explored the psychological mechanisms that underlie this apparent evidence of ‘ghostly’ activity. The experiments took place at two locations that have a considerable reputation for being haunted—Hampton Court Palace (Surrey, England) and the South Bridge Vaults (Edinburgh, Scotland). Both studies involved participants walking around these locations and reporting where they experienced unusual phenomena. Results revealed significantly more reports of unusual experiences in areas that had a reputation for being haunted. This effect was not related to participants' prior knowledge about the reputation of these areas. However, the location of participants' experiences correlated significantly with various environmental factors, including, for example, the variance of local magnetic fields and lighting levels. These findings strongly suggest that alleged hauntings may not necessarily represent evidence for ‘ghostly’ activity, but could be, at least in part, the result of people responding to ‘normal’ factors in their surroundings.
Linearization and return mapping algorithms for elastoplasticity models
Authors: Asensio G. , Moreno C.
Published: May 2003

The return mapping algorithm is one of the most efficient procedures to solve elasto‐plastic problems. However, a criticism that may be lodged against this method is the difficulty of the practical computation of the consistent tangent matrix when the return is non‐radial. Much research has been done to handle this matrix. In this paper, a unified approach is presented in such a way that a simple closed‐form expression gives the consistent tangent matrix for the classical constitutive relations (von Mises, Tresca, Mohr–Coulomb, Drucker–Prager). The basic ideas are in the properties of eikonal equations appearing in several fields as image treatment, short time computation in elastic waves and others. The same kinds of ideas can be extended to non‐classical models. Copyright © 2003 John Wiley & Sons, Ltd.
How “leading quietly” offers realistic help with the difficult, important human problems
Authors: Badaracco Joseph L.
Published: May 2003

Looking at leadership with a wide‐angle lens shows that men and women who do not fit the traditional
heroic mold are, daily, successfully solving important organizational, ethical, and practical problems—and
contributing to a better world. Such quiet leaders operate from a realistic, not cynical, world view based on
four guiding principles: You don't know everything; you will be surprised; keep an eye on the
insiders; and trust, but cut the cards. © 2003 Wiley Periodicals, Inc.
Learning the language of business at Sabre
Authors: Yarbrough Deedra , Rezak Catherine J.
Published: May 2003

Sabre Holdings had to grow up fast. It was spun off from American Airlines in 2000—and then September
11, 2001 slammed the entire airline industry. Yet Sabre came close to achieving its 2002 profitability
plan—in large part because employees controlled expenses. And they had learned both why and how to do that
through an innovative, experiential learning initiative that linked corporate financial analysis with daily work
activity. © 2003 Wiley Periodicals, Inc.
Implementing Lean principles with the Six Sigma advantage: How a battery company realized significant improvements
Authors: Sharma Udit
Published: May 2003

Lean Manufacturing can produce startlingly good results—but implementing them can be costly in time and
other resources, whose investment does not always pay off. When Lean principles are integrated with Six Sigma
practices, however, their success rate grows—and, most importantly, improvements become embedded in daily
work life on a continuing basis. One battery company instituted this blend of methods, and dramatically reduced
its cost of capital while streamlining its manufacturing process—as well as improving its customer
satisfaction levels. © 2003 Wiley Periodicals, Inc.
Practical planning tools that can help get your project off to a good start
Authors: Rieley James B.
Published: May 2003

A variety of difficulties can arise when defining, designing, and coordinating a large project. Some
organizational tools can help each step along the way. © 2003 Wiley Periodicals, Inc.
The zinc finger protein OZF (ZNF146) is overexpressed in colorectal cancer
Authors: Ferbus Didier , Bovin Christophe , Validire Pierre , Goubin Gérard
Published: May 2003

Overexpression of the OZF gene has previously been demonstrated in the majority of pancreatic cancers. However, because the stages of tumour progression in this disease are poorly defined, no conclusion could be drawn concerning the relationship between OZF overexpression and the course of tumour progression. In contrast, initiation and progression steps are well defined in colorectal cancer. Most colon cancers are believed to arise from polypoid adenomas as a result of the gradual accumulation of genetic alterations, allowing the study of genetic events in the early stages of neoplasia. Accordingly, we wanted to assess the frequency of OZF overexpression in this tumour type and the relationship between overexpression and disease stage. Twenty‐five colon carcinoma specimens from different sites and at various stages were analysed by immunoblotting using a highly specific mouse monoclonal antibody. Each sample was compared with adjacent normal colonic mucosa. Complementary immunohistochemical analysis was also carried out on a commercially available tissue array to identify cells expressing OZF. Of the 25 tumours analysed by immunoblotting, 20 expressed higher levels of OZF protein than their adjacent normal mucosa. Immunohistochemistry revealed OZF expression in tumour cells of 56/59 carcinomas and occasionally in infiltrating lymphocytes but at lower levels. Little or no staining was observed in cells taken from normal or inflammatory colon specimens. In both immunoblot and immunohistochemistry experiments, no correlation was found between OZF expression and clinical parameters such as TNM classification, stage, localization and age. Immunostaining was observed in low‐grade adenomas, indicating that OZF expression occurs very early in the course of tumour progression. OZF expression, infrequent or absent in normal colonic mucosa, is present in more than 80% of colorectal cancers. Dysregulation of the OZF gene is an early event that may be implicated in the genesis of colonic carcinoma and may therefore provide a potential therapeutic target. Copyright © 2003 John Wiley & Sons, Ltd.
Prognostic significance of BRCA1 expression in sporadic breast carcinomas
Authors: Lambie H , Miremadi A , Pinder SE , Bell JA , Wencyk P , Paish EC , Macmillan RD , Ellis IO
Published: May 2003

BRCA1 is a tumour suppresser gene frequently mutated in familial breast cancer and thought to influence the progression of sporadic breast cancer. Decreased BRCA1 mRNA and protein expression has been identified in breast cancer cell lines and sporadic breast tumours. Here the prognostic significance of reduced BRCA1 protein expression is investigated in primary operable breast cancer. Immunohistochemical analysis was used to determine the level of BRCA1 protein expression in 100 breast cancers. BRCA1 expression was compared with known prognostic factors and survival to investigate its prognostic significance. BRCA1 nuclear expression was reduced by varying amounts in breast carcinomas. A progressive loss of BRCA1 expression correlated well with higher histological grade (p = 0.002) and an excess of medullary/atypical medullary/grade 3 ductal carcinomas (p = 0.0001). When adjusted for grade, patients with loss of BRCA1 expression had a significantly longer disease‐free survival time. Loss of BRCA1 expression associated with high‐grade breast tumours suggests that BRCA1 may play an important role in the pathogenesis of sporadic breast cancer. Copyright © 2003 John Wiley & Sons, Ltd.
Chronic Granulomatous Mastitis: Diagnostic and Therapeutic Considerations
Authors: Azlina Aziz Firzah , Ariza Zakaria , Arni Talib , Hisham Abdullah Noor
Published: May 2003

Abstract

To review the clinical presentation, histopathological features, and optimal treatment of chronic granulomatous mastitis, the authors conducted a retrospective study of 25 women admitted to a teaching hospital in Malaysia between January 1998 and December 2000 who met the required histologic criteria. The primary outcome measures were morbidity and recurrence of the disease. Thirteen patients presented with a breast mass clinically mimicking breast cancer, and 12 patients had breast induration and abscess formation. In addition, 8 of these patients had recurrent breast disease. Clinical and imaging diagnosis has often been difficult and inconclusive, so histopathology remains the optimal diagnostic tool. Of interest, 50% of patients experience recurrences, and long‐term follow‐up is therefore necessary. The authors concluded that, because chronic granulomatous mastitis is a rare benign breast condition that may be misdiagnosed as breast carcinoma, complete resection should be accomplished whenever possible. Steroid therapy may be an adjuvant for optimal treatment. Awareness among surgeons and pathologists should also be emphasized to avoid unnecessary misdiagnosis and treatment.
New Type of Reconstruction Method after Subtotal Gastrectomy (Noh’s Operation)
Authors: Noh Seung‐Moo , Jeong Hyun‐Yong , Cho June‐Sik , Shin Kyung‐Sook , Song Kyu‐Sang , So Young , Lee Tae‐Yong
Published: May 2003

Abstract

The aim of this study was to compare a new type of reconstruction method (Noh’s operation) with Roux‐en‐Y operation after subtotal gastrectomy. Noh’s operation described herein includes a jejunal occlusion, an end‐to‐side gastrojejunostomy, a side‐to‐end jejunoduodenostomy, and a side‐to‐side jejunojejunostomy after subtotal gastrectomy. A series of 43 patients who had the new operation were compared with 47 patients with the Roux‐en‐Y procedure. The postgastrectomy syndromes, and the mucosal change of the remnant stomach and esophagus were evaluated after surgery. In the new operation, the Roux stasis syndrome occurred in 34.9% at 3 months, in 23.3% at 6 months, in 14.0% at 12 months, and in 11.6% at 24 months. In patients undergoing the Roux‐en‐Y operation, the syndrome occurred in 42.6% at 3 months, in 34.0% at 6 months, in 31.9% at 12 months, and in 29.8% at 24 months. This study shows that the new type of operation (Noh’s operation) can be a good option for reconstruction after subtotal gastrectomy.
Central Pancreatectomy for the Resection of Benign or Low Malignant Potential Neoplasms
Authors: Christein John D. , Kim Anthony W. , Golshan Mehra A. , Maxhimer Justin , Deziel Daniel J. , Prinz Richard A.
Published: May 2003

Abstract

Central pancreatectomy is an uncommonly performed procedure that may be particularly useful for the removal of benign and low malignant potential lesions in the neck and body of the pancreas. This procedure may have fewer major complications and better preserve endocrine and exocrine function than the more commonly performed pancreaticoduodenectomy or distal pancreatectomy. We report our recent experience with central pancreatectomy and review the literature on this topic.
Suspected Acute Deep Vein Thrombosis of the Lower Limb in Outpatients: Considerations for Optimal Diagnostic Approach
Authors: Giannoukas Athanasios D. , Tsetis Dimitrios , Kostas Theodoros , Petinarakis Ioannis , Ioannou Christos , Touloupakis Emmanouel , Katsamouris Asterios N.
Published: May 2003

Abstract

The objective of this study was to review our diagnostic approach using color duplex scanning (CDS) in the management of symptomatic outpatients with suspected lower limb deep venous thrombosis (DVT). CDS was carried out in 315 consecutive outpatients with unilateral symptoms consistent with DVT. Both limbs were assessed in 205 patients. Other pathology was routinely sought when the symptomatic limb was free of thrombosis. Acute DVT was present in 25% (76/315) of the symptomatic limbs, and in only 8% it was confined to calf veins. Other pathology was detected in 90 limbs (28%). Swelling with or without pain was associated with DVT in 44% and 10%, respectively. The time elapsed between the onset of symptoms and CDS was 3 days (range 1–6 days). DVT in the contralateral asymptomatic limb was present in 5 (9%) of the 56 patients with DVT in the symptomatic limb. In the absence of DVT in the symptomatic limb, the contralateral asymptomatic limb was free of thrombosis. Clinical diagnosis of DVT in outpatients was unreliable. CDS revealed that only one‐fourth of the symptomatic limbs had DVT, and other pathology mimicking DVT was present in 28%. CDS is a useful tool that offers a prompt, efficient diagnosis. Investigation of the contralateral asymptomatic limb seems to be necessary only when DVT is found in the symptomatic limb.
Characterization and variation of a human inwardly‐rectifying K‐channel gene (KCNJ6): a putative ATP‐sensitive K‐channel subunit
Authors: Sakura Hiroshi , Bond Chris , Warren-Perry Margaret , Horsley Sharon , Kearney Lyndal , Tucker Stephen , Adelman John , Turner Robert , Ashcroft Frances M.
Published: May 2003

The ATP‐sensitive K‐channel plays a central role in insulin release from pancreatic β‐cells. We report here the cloning of the gene (KCNJ6) encoding a putative subunit of a human ATP‐sensitive K‐channel expressed in brain and β‐cells, and characterisation of its exon‐intron structure. Screening of a somatic cell mapping panel and fluorescent in situ hybridization place the gene on chromosome 21 (21q22.1–22.2). Analysis of single‐stranded conformational polymorphisms revealed the presence of two silent polymorphisms (Pro‐149: CC‐CC and Asp328: GA‐GA) with similar frequencies in normal and non‐insulin‐dependent diabetic patients.
Fibronectin attachment is permissive for IL‐1 mediated gene regulation
Authors: Ostberg Carl O. , Zhu Ping , Wight Thomas N. , Qwarnstrom Eva E.
Published: May 2003

This study examines the effect of cell‐matrix interaction on IL‐1 induced gene regulation. In fibroblasts and smooth muscle cells attached to fibronectin, IL‐1 caused a pronounced reduction in proteoglycan synthesis, while no reduction occurred in cells plated on bare plastic. Further, fibronectin attachment was permissive for IL‐1 mediated suppression of both versican and collagen mRNA levels, initially noted after 4–6 h of IL‐1 incubation. Attachment to vitronectin was less potent in influencing regulation, and collagen had no effect, suggesting specificity of the matrix modulation of the IL‐1 induced response. Similar fibronectin induced dependence was demonstrated for IL‐1 regulation of IL‐6 gene expression, supporting the notion of a general effect of fibronectin receptor engagement on IL‐1 induced signal transduction.
Enzyme‐assisted synthesis of a bivalent high‐affinity dodecasaccharide inhibitor of mouse gamete adhesion The length of the chains carrying distal α1,3‐bonded galactose residues is critical
Authors: Niemelä Ritva , Penttilä Leena , Seppo Antti , Helin Jari , Leppänen Anne , Räbinä Jarkko , Uusitalo Lotta , Maaheimo Hannu , Taskinen Jyrki , Costello Catherine E. , Renkonen Ossi
Published: May 2003

Proposing to study the molecular mechanisms of mouse gamete adhesion with the aid of high affinity adhesion inhibitors of saccharide nature, we report here the enzymatic synthesis of a bivalent oligosaccharide Galα1‐3Galβ1‐4GlcNAcβ1‐3Galβ1‐4GlcNAcβ1‐3(Galα1‐3Galβ1‐4GlcNAcβ1‐3Galβ1‐4GlcNAcβ1‐6)Galβ1‐4GlcNAc (4), consisting of two long arms that link together two distal α1,3‐galactose residues. Binding data reported elsewhere (E. Litscher et al., Biochemistry, 1995, 34, 4662–4669) show that 4 is a high affinity inhibitor of mouse gamete adhesion in vitro (IC50 = 9 μM), while a related octasaccharide Galα1‐3Galβ1‐4GlcNAcβ1‐3(Galα1‐3Galβ1‐4GlcNAcβ1‐6)Galβ1‐4GlcNAc, consisting of two short arms is of very low inhibitory activity. The data highlight the importance of the two α‐galactose residues of 4, and the length of the sugar chains joining them.
Glucose‐induced inactivation of isocitrate lyase in Saccharomyces cerevisiae is mediated by an internal decapeptide sequence
Authors: Ordiz Isabel , Herrero Pilar , Rodicio Rosaura , Moreno Fernando
Published: May 2003

In this work we have investigated the role of specific peptide sequences for glucose‐inactivation of the yeast isocitrate lyase. Thus, different fragments of the ICL1 coding region were fused to the lacZ gene of E. coli to provide a reporter construction. Determinations of β‐galactosidase activities indicated that the decapeptide sequence KTKRNYSARD, located between amino acid residues 37 and 46 of isocitrate lyase, is important for glucose induced proteolytic inactivation. Further experimental evidence was provided by insertion of this sequence into a glucokinase‐β‐alactosidase fusion protein, which is not sensitive to glucose regulation. The decapeptide inserted conferred glucose inactivation to this construct, confirming that it is both necessary and sufficient as a signal.
Post‐transcriptional inhibition of the interleukin‐1 binding protein B15R of Vaccinia virus after coexpression of the related T1 protein
Authors: Danescu Joana , Werenskiold Anne Katrin
Published: May 2003

The interleukin‐1 binding B15R protein of Vaccinia virus and murine T1 are related extracellular glycoproteins with similarity to the extracellular domain of interleukin‐1 receptors. In cells infected with a recombinant Vaccinia virus directing the overexpression of T1, production of the endogenous viral B15R protein is abrogated. T1 synthesis specifically interferes with the production of B15R, but not of other secretory viral proteins. Inhibition of B15R expression occurs at the posttranscriptional level, is exerted in trans and requires the presence of T1 protein in the infected cell. These results suggest a common maturation pathway for the B15R and T1 proteins which might also apply to other members of the interleukin‐1 receptor family.
A kinetic model for comparing proteolytic processing activity and inhibitor resistance potential of mutant HIV‐1 proteases
Authors: Tang Jordan , Hartsuck Jean A.
Published: May 2003

A kinetic model is presented for the comparison of potential proteolytic processing activities of wild‐type and mutant human immunodeficiency virus proteases in the presence and absence of protease inhibitors. The protease processing of gag substrate in the immature virions is assessed by the kinetic parameters, k
cat, K
m and K
i. The relationship of the estimated potential processing activities to the viability of the HIV mutants and their tendencies to resist inhibitors also are discussed. A fully developed model has the potential to simulate the results of inhibitor resistance either in vivo or in cell culture.
Multiplicity of N‐terminal structures of medium‐chain alcohol dehydrogenases Mass‐spectrometric analysis of plant, lower vertebrate and higher vertebrate class I, II, and III forms of the enzyme
Authors: Hjelmqvist Lars , Hackett Murray , Shafqat Jawed , Danielsson Olle , Iida Junko , Hendrickson Ronald C. , Michel Hanspeter , Shabanowitz Jeffrey , Hunt Donald F. , Jönvall Hans
Published: May 2003

Ten different alcohol dehydrogenases, representing several classes of the enzyme and a wide spread of organisms, were analyzed for patterns of N‐terminal structures utilizing a combination of conventional and mass spectrometric peptide analysis. Results show all forms to be N‐terminally acetylated and allow comparisons of now 40 such alcohol dehydrogenases covering a large span of forms and origins. Patterns illustrate roles of acetylation in proteins in general, define special importance of the class I N‐terminal acetylation, and distinguish separate acetylated structures for all classes, as well as a common alcohol dehydrogenase motif.
The mutant Asn291 → Ser human lipoprotein lipase is associated with reduced catalytic activity and does not influence binding to heparin
Authors: Buscà Roser , Peinado Julia , Vilella Elisabeth , Auwerx Johan , Deeb Samir S , Vilaró Senén , Reina Manuel
Published: May 2003

Lipoprotein lipase (LPL) plays a central role in triglyceride metabolism, regulating the catabolism of triglyceride‐rich lipoprotein particles. LPL performs its hydrolytic action attached to heparan sulfate proteoglycans at the luminal surface of capillary endothelial cells. We have assessed the effect of the Asn291 → Ser (N291S) substitution found in LPL gene from a human hyperlipemic patient. Our results showed that both the wild‐type (WT) and N291S hLPL expressed in COS1 cells were secreted to the extracellular medium, and presented similar intracellular distibution patterns. Furthermore, heparin‐Sepharose affinity chromatography assays revealed normal heparin affinity of the N291S hLPL. In addition, both the mutant and the WT protein bound to the surface of human fibroblasts and untransfected COS1 cells. Interestingly, disminished LPL specific activity was observed in the extracellular medium from mutant expressing cells. Therefore the lack of normal LPL activity in patients harbouring such a mutation could be the cause of their hyperlipemic disorder.
Trigger factor, one of the Escherichia coli chaperone proteins, is an original member of the FKBP family
Authors: Callebaut Isabelle , Mornon Jean-Paul
Published: May 2003

The trigger factor of Escherichia coli is known as a chaperone protein which forms soluble complexes with the precursor to outer membrane protein A and assists in the maintenance of translocation competence. Sequence analysis shows that trigger factor contains a domain belonging to the FK506‐binding protein (FKBP) family and possessing all the amino acids necessary for FK506 binding and peptidyl‐prolyl cis‐trans isomerase (Ppiase) activity. Consequently, this protein could be directly involved in the unfolding/folding processes occurring during translocation across the E. coli plasma membrane and, more generally, in facilitating protein folding. The central position of the FKBP domain within the trigger factor sequence as well as several original features of the loops surrounding the FK506‐binding pocket are not found in any other FKBPs, making it undetectable by the Fkbp‐Ppiase signature patterns.
An ICE‐like protease is a common mediator of apoptosis induced by diverse stimuli in human monocytic THP.1 cells
Authors: Zhu Huijun , Fearnhead Howard O. , Cohen Gerald M.
Published: May 2003

Apoptosis was induced in THP.1 cells, a human monocytic tumour cell line, by diverse stimuli including cycloheximide, thapsigargin, etoposide and staurosporine. Induction of apoptosis by all these stimuli, except etoposide, was enhanced in the presence of the trypsin‐like protease inhibitor, chloromethyl ketone (TLCK). Induction of apoptosis, assessed by morphological, flow cytometric and biochemical criteria, including proteolysis of poly(ADP‐ribose) polymerase and cleavage of DNA to large kilobasepair fragments, was completely abrogated when cells were pretreated with an ICE‐like protease inhibitor, ZValAlaAsp·fluoromethylketone. This suggested that an ICE homologue was a common mediator of apoptosis in THP.1 cells.
Point mutation of a conserved arginine (104) to lysine introduces hypersensitivity to inhibition by glyphosate in the 5‐enolpyruvylshikimate‐3‐phosphate synthase of Bacillus subtilis
Authors: Selvapandiyan Angamuthu , Majumder Kumud , Fattah Farkad A. , Ahmad Suhail , Arora Naresh , Bhatnagar Raj K.
Published: May 2003

The role of a conserved arginine (R104) in the putative phosphoenol pyruvate binding region of 5‐enolpyruvyl shikimate‐3‐phosphate synthase of Bacillus subtilis has been investigated. Employing site directed mutagenesis arginine was substituted by lysine or glutamine. Native and mutant proteins were expressed and purified to near homogeneity. Estimation of Michaelis and inhibitor constants of the native and mutant proteins exhibited altered substrate—inhibitor binding mode and constants. Mutation R104K hypersensitized the enzyme reaction to inhibition by glyphosate. The role of R104 in discriminating between glyphosate and phosphoenol pyruvate is discussed.
Deficient DNA repair of triple helix‐directed double psoralen damage in human cells
Authors: Sandor Zoltan , Bredberg Anders
Published: May 2003

Damage induced by a single psoralen‐modified triple helix‐forming oligonucleotide has been reported to be efficiently repaired in human cells. In this study we investigated a set of psoralen coupled oligonucleotides introducing multiple lesions into the target DNA. A simian virus 40 (SV40) shuttle vector was in vitro treated with different triple helix‐forming oligonucleotides and UVA radiation, leading to double psoralen adducts at the supF mutational target gene of the plasmid. After passage in the Raji human cell line the recovered vector was analysed in an indicator bacterial strain. The results show that double psoralen adducts, located at both ends of a long triple helix, cannot be repaired efficiently in human cells.
Thermal stabilization of a single‐chain Fv antibody fragment by introduction of a disulphide bond
Authors: Young N.Martin , MacKenzie C.Roger , Narang Saran A. , Oomen Raymond P. , Baenziger John E.
Published: May 2003

A disulphide bond was introduced into a single‐chain Fv form of the anticarbohydrate antibody, Se155‐4 by replacing Ala‐L57 of the light chain and Asp‐H106 of the heavy chain with cysteines, by site‐directed mutagenesis. To maintain the saltbridge from the latter residue to Arg‐H98, Tyr‐107 was also altered to Asp. The resulting ds‐scFv was shown to retain full antigen‐binding activity, by enzyme immunoassay and surface plasmon resonance analysis of binding kinetics. Compared with the parent scFv, the disulphide bonded form was shown to have enhanced thermal stability, by Fourier transform IR spectroscopy. The T
m was raised from 6°C to 69°C. The ds‐scFv form thus combines the stable monomeric form of the disulphide form with the expression advantages of the scFv.
Synthesis, cloning and expression in Escherichia coli of a gene coding for the Met8 → Leu CMTI I — a representative of the squash inhibitors of serine proteinases
Authors: Bolewska Krystyna , Krowarsch Daniel , Otlewski Jacek , Jaroszewski Lukasz , Bierzynski Andrzej
Published: May 2003

A chemically synthesized gene coding for a Cucurbita maxima trypsin inhibitor modified at position P′3 (Met8→ Leu CMTI I), i.e. at the third position downstream of the reactive site bond (Arg5‐Ile), was cloned into a derivative of the plasmid pAED4 that utilizes a T7 expression system. The gene was expressed in Escherichia coli as a fusion protein that accumulates in inclusion bodies. After reduction and CNBr cleavage of the fusion protein followed by oxidative refolding and reverse‐phase HPLC, about 5 mg of pure protein was obtained per 1 of cell culture. Association constants of recombinant Leu‐8‐CMTI I with bovine β‐trypsin and human cathepsin G are the same, within experimental error, as for CMTI I isolated from a natural source.
A novel approach for expression cloning of small GTPases: identification, tissue distribution and chromosome mapping of the human homolog of rheb
Authors: Gromov Pavel S. , Madsen Peder , Tomerup Niels , Celis Julio E.
Published: May 2003

We report a novel approach for identifying monomeric GTP‐binding proteins that is based on probing cDNA expression libraries with [α‐32P]GTP. In short, a nitrocellulose replica from a plated cDNA expression library is treated with 2% SDS to block the GTP‐binding activity of various G proteins expressed by E. coli, thus allowing the direct identification of positive clones. Using this procedure we have cloned several small GTP‐binding proteins from human keratinocytes including the human homolog of rheb, a novel member of the ras‐related GTP‐binding proteins. Human rheb cDNA shares 90% identity with the rat counterpart and it is highly upregulated in transformed human cells of various origin. Northern analysis showed that human rheb is ubiquitously expressed, with the highest levels observed in skeletal and cardiac muscle, and not in brain, as it is the case for rat rheb. The human RHEB gene was mapped to chromosome 10q11.
Electrophysiological properties of the hypokalaemic periodic paralysis mutation (R528H) of the skeletal muscle α1S subunit as expressed in mouse L cells
Authors: Lapie Pascale , Goudet Cyril , Nargeot Joël , Fontaine Bertrand , Lory Philippe
Published: May 2003

Hypokalaemic periodic paralysis (HypoPP) is an autosomal dominant muscle disease which has been linked to point mutations in the skeletal muscle L‐type calcium channel α
1 subunit (α
1S
). Here, we have introduced one of the point mutations causing HypoPP (R528H) into cDNA of the rabbit α
1S
. Expression of either the wild‐type α
1S or the mutant R528H α
1S
(α
1
S−R528H) subunit was obtained in mouse Ltk− cell using a selectable expression vector. The α
1S−R528H
subunit led to the expression of functional L‐type Ca2+ channels. Corresponding whole‐cell Ba2+ currents exhibit very slow activation and inactivation kinetics, typical for recombinant skeletal Ca2+ channel currents. Voltage‐dependent activation and inactivation properties were similar for α
1S
‐and α
1S − R528H
, as well as their sensitivity to the dihydropyridine agonist Bay K 8644. Differences in α
1S
‐and α
1S−R528H
‐directed channels reside in the Ba2+ current density, which was significantly reduced 3.2 fold in cells expressing α
1S − R528H
. It was concluded that the R528H mutation of α
1S
results in minor differences in the electrophysiological properties but significantly reduces the whole‐cell Ca2+ channel current in its amplitude.
Tannin interactions with a full‐length human salivary proline‐rich protein display a stronger affinity than with single proline‐rich repeats
Authors: Charlton Adrian J. , Baxter Nicola J. , Lilley Terence H. , Haslam Edwin , McDonald Charles J. , Williamson Michael P.
Published: May 2003

The protein IB5 has been purified from human parotid saliva. This protein contains several repeats of a short proline‐rich sequence. Dissociation constants have been measured at several discrete binding sites using 1H‐NMR for the hydrolysable tannins (polyphenols) (, and and the condensed proanthocyanidin (−)‐epicatechin. The dissociation constants for trigalloyl glucose and pentagalloyl glucose were 15 × 10−5 and 1.7 × 10−5 M, respectively, which are 115 and 1660 times stronger than those previously measured under the same conditions for a single repeat of a mouse salivary proline‐rich protein. The increase in affinity is ascribed to intramolecular secondary interactions, which are strengthened by the rigidity of the interacting molecules.
Pseudogenes in ribonuclease evolution: a source of new biomacromolecular function?
Authors: Trabesinger-Ruef Nathalie , Jermann Thomas , Zankel Todd , Durrant Barbara , Frank Gerhard , Benner Steven A.
Published: May 2003

Bovine seminal ribonuclease (RNase) diverged from pancreatic RNase after a gene duplication ca. 35 million years ago. Members of the seminal RNase gene family evidently remained as unexpressed pseudogene for much of its evolutionary history. Between 5 and 10 million years ago, however, after the divergence of kudu but before the divergence of ox, evidence suggests that the pseudogene was repaired and expressed. Intriguingly, detailed analysis of the sequences suggests that the repair may have involved gene conversion, transfer of information from the pancreatic gene to the RNase pseudogene. Further, the ratio of non‐silent to silent substitutions suggests that the pancreatic RNases are divergently evolving under functional constraints, the seminal RNase pseudogenes are diverging under no functional constraints, while the genes expressed in the seminal plasma are evolving extremely rapidly in their amino acid sequences, as if to fulfil a new physiological role.
Monitoring process parameters with pre‐control
Authors: Olorunniwo Festus , Knight John Ed
Published: May 2003

Many manufacturers have reported success in using Pre‐Control (PC) procedures to monitor their processes. The reported success has been attributed to the fact that the operators, who usually implement PC procedure, find it simple. Beside simplicity, PC has several other advantages over conventional control charts ( charts, etc.). PC ensures operator control of a process as well as product quality, an essential factor in operator empowerment. The procedure can also be effectively used on short production runs; it ensures quick process qualification; provides quick feedback of information on a process; and is very economical to operate because it requires minimum paperwork and mathematical calculations.

The simplicity of use is attributed to the fact that, in its easiest and most common mode of implementation, PC requires no numerical recording or scaled plotting of data. For these same reasons it has been difficult, in this mode of implementation, to evaluate the values of the process parameters (mean and variability). In the other mode of PC implementation where numerical recording and plotting of data is routinely required, the simplicity advantage is lost. Because of the inability to evaluate the process parameters, manufacturers using the common mode of PC implementation are frustrated because they are unable to demonstrate that their processes are capable of, and are in fact producing, products of high quality.

In this paper we propose two models for evaluating process parameters without the need to record any numerical measurement of product characteristics. Although the two models are jointly executed, the first evaluates process variability, and the second process mean. Common PC procedure normally uses indicator gages with zones colored identically as the zones of the ‘charts’. We show that the only additional requirement to execute the models is to subdivide each of the four zones on the gage into two. The ‘chart’ itself needs not be drawn to scale. The operator can draw the required lines on the ‘chart’ free‐hand. Points representing zones indicated by the gage's “measurements” are marked anywhere in a “box” on the chart without the need for accurately locating the points.

We conducted a simulation analysis to evaluate the accuracy of our models (measured by the absolute percent error of the estimates of the process parameters from the actual). We report that our models perform favorably compared with traditional measures of variability and mean. For manufacturers who use the simple PC procedures we believe that the proposed models now provide a means to also quantify the process capability.
Minimum tardiness scheduling in flow shops: Construction and evaluation of alternative solution approaches
Authors: Raman Narayan
Published: May 2003

Surveys of industrial scheduling practice show that meeting customer due dates is a critical concern for many manufacturing systems. While there is considerable research on the effectiveness of scheduling rules in job shops, very little work is reported on flow shops. Although scheduling rules developed for job shops can be applied in other systems as well, we show that the inherent structure of a flow shop can be utilized to construct effective solution procedures. In particular, for the total tardiness problem, we develop conditions for local optimality in a 2‐machine flow shop, and use these results to generate an efficient improvement heuristic procedure. We also construct solution methods that utilize the notion of shifting bottlenecks.

Our computational experience reveals the superiority and robustness of the proposed approaches under a variety of problem scenarios. The results of this study also indicate the need to look beyond permutation schedules for solving the flow shop tardiness problem. Furthermore, they suggest that the criterion for determining machine criticality needs to take the scheduling objective into consideration. While it appears intuitive to impute criticality on the basis of machine workloads and, therefore, emphasize the bottleneck machine for generating the shop schedule, this approach may frequently result in inferior system performance.
POM research productivity in U.S. business schools
Authors: Young Scott T. , Baird Brad C. , Pullman Madeleine E.
Published: May 2003

This study examines the publication output of individuals and business schools in the field of Production and Operations Management (POM). A ranking system was devised based upon publications in the leading POM journals. Two ranking approaches were applied to business schools: productivity and quality. Articles were counted in two ways: one ranking counted each article as one unit, and authors “shared” the unit equally, and the second ranking gave each contributor credit for one “distributed” unit.

These are the first publication productivity rankings for the field of POM, and are based upon publications in the years 1989–1993.
Characterization of thioredoxin y, a new type of thioredoxin identified in the genome of Chlamydomonas reinhardtii
Authors: Lemaire S.D. , Collin V. , Keryer E. , Quesada A. , Miginiac-Maslow M.
Published: May 2003

The sequencing of the Arabidopsis genome revealed a multiplicity of thioredoxins (TRX), ubiquitous protein disulfide oxido‐reductases. We have analyzed the TRX family in the genome of the unicellular green alga Chlamydomonas reinhardtii and identified eight different thioredoxins for which we have cloned and sequenced the corresponding cDNAs. One of these TRXs represents a new type that we named TRX y. This most probably chloroplastic TRX is highly conserved in photosynthetic organisms. The biochemical characterization of the recombinant protein shows that it exhibits a thermal stability profile and specificity toward target enzymes completely different from those of TRXs characterized so far.
Channel induction by palytoxin in yeast cells expressing Na+,K+‐ATPase or its chimera with sarco/endoplasmic reticulum Ca2+‐ATPase
Authors: Ito Katsuaki , Toyoda Isao , Higashiyama Masato , Uemura Daisuke , Sato Masa H. , Yoshimura Shige H. , Ishii Toshiaki , Takeyasu Kunio
Published: May 2003

Palytoxin (PTX) induces a cation channel through interaction with Na+,K+‐ATPase. It is unclear how this action relates to the enzyme catalytic activity. We examined whether the action of PTX depends on the catalytic domain specific for Na+,K+‐ATPase. Wild‐type Na+,K+‐ATPase α‐subunit (NNN) or its chimera (NCN), in which the catalytic domain was replaced with that of sarcoplasmic/endoplasmic reticulum Ca2+‐ATPase, was co‐expressed with β‐subunit in the yeast Saccharomyces cerevisiae. PTX (0.1–100 nM) increased K+ efflux in NNN‐ or NCN‐transfected cells to a similar degree but not in non‐transfected cells. When ouabain‐resistant NNN and NCN were expressed, PTX also increased K+ efflux. Ouabain inhibited the effect of PTX in NNN or NCN cells but not in ouabain‐resistant cells. These data suggest that the channel‐forming action of PTX does not depend on the catalytic domain species.
Complementation of Escherichia coli ubiF mutation by Caenorhabditis elegans CLK‐1, a product of the longevity gene of the nematode worm
Authors: Adachi Akihiko , Shinjyo Noriko , Fujita Daisuke , Miyoshi Hideto , Amino Hisako , Watanabe Yoh-ichi , Kita Kiyoshi
Published: May 2003

Caenorhabditis elegans CLK‐1 was identified from long‐lived mutant worms, and is believed to be involved in ubiquinone biosynthesis. The protein belongs to the eukaryotic CLK‐1/Coq7p family, which is also similar to the bacterial Coq7 family, that hydroxylates demethoxyubiquinone, resulting in the formation of hydroxyubiquinone, a precursor of ubiquinone. In Escherichia coli, the corresponding reaction is catalyzed by UbiF, a member of a distinct class of hydroxylase. Although previous studies suggested that the eukaryotic CLK‐1/Coq7 family is a hydroxylase of demethoxyubiquinone, there was no direct evidence to show the enzymatic activity of the eukaryotic CLK‐1/Coq7 family. Here we show that the plasmid encoding C. elegans CLK‐1 supported aerobic respiration on a non‐fermentable carbon source of E. coli ubiF mutant strain and rescued the ability to synthesize ubiquinone, suggesting that the eukaryotic CLK‐1/Coq7p family could function as bacterial UbiF.
Ca2+‐induced oxidative stress in brain mitochondria treated with the respiratory chain inhibitor rotenone
Authors: Sousa Solange C. , Maciel Evelise N. , Vercesi Anibal E. , Castilho Roger F.
Published: May 2003

In this study we show that micromolar Ca2+ concentrations (>10 μM) strongly stimulate the release of reactive oxygen species (ROS) in rotenone‐treated isolated rat forebrain mitochondria. Ca2+‐stimulated mitochondrial ROS release was associated with membrane lipid peroxidation and was directly correlated with the degree of complex I inhibition by rotenone. On the other hand, Ca2+ did not increase mitochondrial ROS release in the presence of the complex I inhibitor 1‐methyl‐4‐phenylpyridinium. Cyclosporin A had no effect on Ca2+‐stimulated mitochondrial ROS release in the presence of rotenone, indicating that mitochondrial permeability transition is not involved in this process. We hypothesized that Ca2+‐induced mitochondrial oxidative stress associated with partial inhibition of complex I may be an important factor in neuronal cell death observed in the neurodegenerative disorder Parkinson's disease.