bluuebunny/crossref_metadata_2025 huggingface dataset:
Innovative and sustainable energy technologies: the role of venture capital
Authors: Moore Bill , Wüstenhagen Rolf
Published: July 2004

This paper provides an overview of a multi‐year research project being conducted at the University of St. Gallen, Switzerland. The objective of the research is to evaluate the role of financial structure in promoting the development of innovative and sustainable energy technologies and its current relationship to innovation and entrepreneurial activity in this important economic sector. In this paper, emphasis is placed on the venture capital segment of the capital market and the electricity segment of the energy industry. We first provide a short overview of the electricity sector. We then briefly discuss the concept of innovation and entrepreneurship and its relationship to our research effort. Finally, the bulk of the paper outlines the conceptual framework for the venture capital element of the research project. We conclude the paper with a summary of our preliminary observations. Copyright © 2004 John Wiley & Sons, Ltd and ERP Environment.
Eight types of product–service system: eight ways to sustainability? Experiences from SusProNet
Authors: Tukker Arnold
Published: July 2004

Function‐oriented business models or product–service systems (PSSs) are often seen as an excellent means for achieving ‘factor 4’. SusProNet, an EU network on PSSs, showed a more complicated reality. At least eight different types of PSS exist, with quite diverging economic and environmental characteristics. The economic potential of each type was evaluated in terms of (i) tangible and intangible value for the user, (ii) tangible costs and risk premium for the provider, (iii) capital/investment needs and (iv) issues such as the providers' position in the value chain and client relations. The environmental potential was evaluated by checking the relevance of certain impact reduction mechanisms (e.g. more intensive use of capital goods, inherent incentives for sustainable user and provider behaviour etc.). Most PSS types will result in marginal environmental improvements at best. The exception is the PSS type known as functional results, but here liability and risk premium issues, amongst others, need a solution. Copyright © 2004 John Wiley & Sons, Ltd and ERP Environment.
Synthesis of Tailor‐Made Glycoconjugate Mimetics of Heparan Sulfate That Bind IFN‐γ in the Nanomolar Range
Authors: Lubineau André , Lortat‐Jacob Hugues , Gavard Ollivier , Sarrazin Stéphane , Bonnaffé David
Published: July 2004

We have recently described the preparation of three building blocks for the combinatorial synthesis of heparan sulfate (HS) fragments. Herein we show that one of these building blocks (disaccharide 4) allows the preparation, in high yields and with total α stereoselectivity, of tetra‐, hexa‐ and octasaccharides from the heparin (HP) regular region, by using 2+2, 2+4 and 4+4 glycosylation strategies, respectively. These oligosaccharides were processed into sulfated derivatives bearing an allyl moiety in the anomeric position. The UV‐promoted conjugation of these compounds with α,ω‐bis(thio)poly(ethylene glycol) spacers of three different lengths allowed us to prepare nine benzylated glycoconjugates. After final deprotection, the glycoconjugates 1 a–c, 2 a–c and 3 a–c were obtained and their ability to inhibit the interaction between IFN‐γ and HP was tested by using surface plasmon resonance detection. Compound 3 b, containing two HP octasaccharides linked by a 50‐Å linker was able to inhibit the IFN‐γ/HP interaction with an IC50 value of approximately 35 nM. In addition, the nine glycoconjugates were perfect tools in the study to ascertain the topology of the IFN‐γ binding site on HS. Compounds 1 a–c, 2 a–c and 3 a–c, by mimicking the alternating sulfated and nonsulfated regions found in HS, thus comprise the first example of a library of synthetic HS mimetics giving access to the “second level of molecular diversity” found in HS.
Adrenaline Recognition in Water
Authors: Molt Oliver , Rübeling Daniel , Schäfer Gerhard , Schrader Thomas
Published: July 2004

Host molecule 1 displays a high affinity in water towards catecholamines and especially related structures such as β‐blockers with extended aromatic π‐faces (up to 7×103 M−1 for each single complexation step or 5×107 M−2 for both steps). The amphiphilic structural design leads to an extensive self‐association of host molecules through their aromatic flanks. Above a cmc (critical micelle concentration) of 3×10−4 M, host 1 forms micelles that produce a favorable microenvironment for hydrophobic interactions with the included guest molecules. Electrostatic attraction of the ammonium alcohol by the phosphonate anions is thus combined with hydrophobic contributions between the aromatic moieties. Ionic hydrogen bonds with polar OH or NH groups of the guest enforce the non‐covalent interactions, and finally lead to increased specificity. Both its affinity and its selectivity towards adrenergic receptor substrates are greatly enhanced if the receptor molecule 1 is transferred from water into a lipid monolayer. Catecholamines and β‐blockers lead to drastically different effects at concentrations approaching the micromolar regime. Especially β‐blockers with minute structural changes can be easily distinguished from each other. In both cases, extensive hydrophobic interactions with a self‐associated and/or self‐organized microenvironment are largely responsible for the observed high efficiency and specificity.
Synthesis of a Polymer‐Bound Galactosylamine and Its Application as an Immobilized Chiral Auxiliary in Stereoselective Syntheses of Piperidine and Amino Acid Derivatives
Authors: Zech Gernot , Kunz Horst
Published: July 2004

A 2,3,4‐tri‐O‐pivaloylated β‐D‐galactopyranosyl azide bearing a hydroxy‐functionalized spacer unit at the C‐6 position of the galactose was synthesized and immobilized on the solid phase by using a polymer‐bound chlorosilane. The azide was reduced to the corresponding galactopyranosylamine, which served as a versatile chiral auxiliary in highly diastereoselective Ugi four‐component condensation reactions at ambient temperature. Fluoride‐induced cleavage from the polymeric support furnished N‐glycosylated N‐acylated α‐amino acid amides. The reaction of the immobilized galactosylamine with aldehydes gave rise to the corresponding aldimines, which underwent a domino Mannich–Michael condensation reaction with Danishefsky's diene at ambient temperature to yield 2‐substituted 5,6‐didehydropiperidin‐4‐ones on the solid phase. Subsequent cleavage with tetra‐n‐butylammonium fluoride delivered the N‐glycosylated products in high yields, purities, and diastereoselectivities. A chemoselective 1,4‐hydride addition to the polymer‐bound dehydropiperidinones was achieved in the presence of the bulky oxygenophilic Lewis acid methylaluminum [bis(2,6‐di‐tert‐butyl‐4‐methylphenoxide)]. The conjugate addition of cyano‐modified Gilman reagents to the immobilized dehydropiperidinones furnished 2,6‐cis‐substituted piperidine derivatives as the major diastereomers that were isolated after cleavage from the support.
Green fluorescent protein fusions to Arabidopsis fimbrin 1 for spatio‐temporal imaging of F‐actin dynamics in roots
Authors: Wang Yuh‐Shuh , Motes Christy M. , Mohamalawari Deepti R. , Blancaflor Elison B.
Published: July 2004

The visualization of green fluorescent protein (GFP) fusions with microtubule or actin filament (F‐actin) binding proteins has provided new insights into the function of the cytoskeleton during plant development. For studies on actin, GFP fusions to talin have been the most generally used reporters. Although GFP‐Talin has allowed in vivo F‐actin imaging in a variety of plant cells, its utility in monitoring F‐actin in stably transformed plants is limited particularly in developing roots where interesting actin dependent cell processes are occurring. In this study, we created a variety of GFP fusions to Arabidopsis Fimbrin 1 (AtFim1) to explore their utility for in vivo F‐actin imaging in root cells and to better understand the actin binding properties of AtFim1 in living plant cells. Translational fusions of GFP to full‐length AtFim1 or to some truncated variants of AtFim1 showed filamentous labeling in transient expression assays. One truncated fimbrin‐GFP fusion was capable of labeling distinct filaments in stably transformed Arabidopsis roots. The filaments decorated by this construct were highly dynamic in growing root hairs and elongating root cells and were sensitive to actin disrupting drugs. Therefore, the fimbrin‐GFP reporters we describe in this study provide additional tools for studying the actin cytoskeleton during root cell development. Moreover, the localization of AtFim1‐GFP offers insights into the regulation of actin organization in developing roots by this class of actin cross‐linking proteins. Cell Motil. Cytoskeleton 59:79–93, 2004. © 2004 Wiley‐Liss, Inc.
Cancer patients' expectations of experiencing treatment‐related side effects
Authors: Hofman Maarten , Morrow Gary R. , Roscoe Joseph A. , Hickok Jane T. , Mustian Karen M. , Moore Dennis F. , Wade James L. , Fitch Tom R.
Published: July 2004

BACKGROUND

Adequate management of treatment‐related side effects is important for patients and challenging for clinicians. Side effects generated by various treatments have been characterized reasonably well. However, to the authors' knowledge, less is known regarding what patients expect to experience regarding these side effects and how patient characteristics are related to these expectations.

METHODS

Patients with cancer (n = 1015 patients) from 17 Community Clinical Oncology Program (CCOP) institutions affiliated with the University of Rochester Cancer Center CCOP Research Base were surveyed regarding their expectations of experiencing side effects associated with cancer treatment, with 938 patients providing evaluable data. Patients responded to the item, “Indicate your expectations of having this side effect” for 12 common side effects. Patients rated their expectations using a 5‐point Likert scale, from 1 (“I definitely will not have this”) to 5 (“I definitely will have this”).

RESULTS

The median number of symptoms expected (characterized by any value other than one) was nine. The six most expected symptoms were fatigue, nausea, sleep disturbance, weight loss, hair loss, and skin problems. Patients age > 60 years expected to have fewer symptoms than younger patients; female patients expected more side effects than male patients; and patients who had some college education expected more side effects than patients who were high school graduates or had not completed high school.

CONCLUSIONS

Patients with cancer clearly exhibit expectations regarding treatment‐related side effects; and age, gender, and education level appear to influence these expectations. Further careful characterization of patient expectations and how expectations relate to experience may lead to earlier and more effective management of side effects. Cancer 2004. © 2004 American Cancer Society.
MammoSite and interstitial brachytherapy for accelerated partial breast irradiation
Authors: Shah Neenad M. , Tenenholz Todd , Arthur Douglas , DiPetrillo Thomas , Bornstein Bruce , Cardarelli Gene , Zheng Zhen , Rivard Mark J. , Kaufman Seth , Wazer David E.
Published: July 2004

BACKGROUND

In interstitial brachytherapy (IB), cosmesis and toxicity correlate with volume of tissue irradiated, dose homogeneity index (DHI), and adjuvant doxorubicin/cyclophosphamide based chemotherapy (ACCT). MammoSite brachytherapy (MSB) irradiates smaller volumes than IB, and lower dose homogeneity does not appear to affect toxicity. However, clinical experience suggests that other factors may also play an important role in cosmesis and toxicity with MSB. We reviewed our prospectively maintained data base of women who underwent accelerated partial breast irradiation (APBI) to assess this issue.

METHODS

Beginning in September 1995, 115 women were enrolled in a trial evaluating APBI as monotherapy after lumpectomy. The first 75 eligible patients received IB, and the most recent 28 eligible patients received MSB. All patients received 34 gray (Gy) in 10 twice‐daily fractions through high‐dose rate iridium‐192 brachytherapy; 19% of patients in the IB group and 0% of patients in the MSB group received ACCT.

RESULTS

At 1 year after treatment, MSB caused significantly less Grade 2–4 subcutaneous fibrosis (as graded by a radiation oncologist according to the Radiation Therapy Oncology Group/Eastern Cooperative Oncology Group system) compared with IB (10.7% vs. 32%; P = 0.04). However, when only ACCT‐naïve patients in the IB group were compared with patients in the MSB group, this finding became nonsignificant. Among the patients who received MSB, significantly smaller volumes were irradiated, and the DHI was lower.

CONCLUSIONS

Current studies suggest an improved toxicity profile with MSB compared with IB that is attributed to lower irradiated volumes with MSB. When only chemotherapy‐naïve patients were compared, however, toxicity and cosmesis were found to be similar between MSB and IB, suggesting a more complex interplay between irradiated volumes, DHI, and chemotherapy. The relation of ACCT to toxicity in this scenario is intriguing and warrants further investigation. Cancer 2004. © 2004 American Cancer Society.
Estimation of an optimal radiotherapy utilization rate for gynecologic carcinoma
Authors: Delaney Geoff , Jacob Susannah , Barton Michael
Published: July 2004

BACKGROUND

Radiotherapy usage rates exhibit wide variations both within and between countries. Current estimates of the proportion of cancer patients who should optimally receive radiotherapy are based either on expert opinion or on the measurement of actual usage rates rather than on the best available scientific evidence.

METHODS

With the goal of developing an evidence‐based benchmark for radiotherapy use in the treatment of malignancies of the cervix, vagina, vulva, and ovary (endometrial malignancies are covered in a separate article), the authors reviewed international evidence‐based treatment guidelines. Optimal radiotherapy usage trees were constructed, and proportions of patients with clinical indications for radiotherapy were obtained from epidemiologic data. These ideal usage rates were compared with actual radiotherapy utilization rates recorded in Australia and elsewhere.

RESULTS

According to the best available evidence, radiotherapy is indicated at least once for 58% of patients with cervical carcinoma, 4% of patients with ovarian carcinoma, 100% of patients with vaginal carcinoma, and 34% of patients with vulvar carcinoma. A review of the limited data available suggests that actual radiotherapy usage rates for patients with gynecologic malignancies are comparable to optimal usage rates.

CONCLUSIONS

Actual practice appears to approximate the authors' model of optimal radiotherapy use. This finding reflects the high level of agreement among treatment guidelines as well as the existence of high‐quality evidence related to the management of gynecologic malignancies, and it may also be indicative of the fact that a large proportion of patients are treated in specialist units. The management of gynecologic malignancies may serve as a good example in the development of management strategies for other types of cancer. Cancer 2004. © 2004 American Cancer Society.
Towards Cationic Gallium Derivatives: Metallacycles from the Reactions of Organogallium Compounds with Tetraorganodichalcogenoimidodiphosphinates and a New N‐(Diphenylthiophosphinyl)thioureato Ligand
Authors: Montiel‐Palma Virginia , Huitrón‐Rattinger Estefanía , Cortés‐Llamas Sara , Muñoz‐Hernández Miguel‐Ángel , García‐Montalvo Verónica , López‐Honorato Eddie , Silvestru Cristian
Published: July 2004

The organometallic complexes of general formulae [Me2Ga{η2‐E,E′‐[R2P(E)NP(E′)R′2]}] [R = R′ = Ph, E = E′ = O (1); R = R′ = Ph, E = E′ = S (2); R = R′ = Ph, E = E′ = Se (3); R = R′ = Ph, E = O, E′ = S (4); R = Me, R′ = Ph, E = S, E′ = O (5)] and [Me2Ga{η2‐S,S′‐[Ph2P(S)NC(S)(C9H10N)]}] (6) were obtained by facile methane elimination reactions from GaMe3 and the acidic ligands L1H [(XPPh2)2NH (X = O, S, Se), (OPPh2)(SPPh2)NH, and (OPMe2)(SPPh2)NH] and L2H [Ph2P(S)NHC(S)(C9H10N)] in toluene. Replacement of one phosphorus atom by a carbon atom in the ligand skeleton of L1H gave the new ligand L2H, which, upon reaction with GaMe3, gave compound 6, which shows no significant structural differences with respect to 1−5. Therefore, L2H does not induce partial planarity in the six‐membered ring, indicating the necessity for replacing both phosphorus atoms of the ligand by carbon atoms, as in the β‐diketonate‐type derivatives, in order to impose ring planarity. Thus, despite originating from a variety of ligands with differing donor atoms and substituents at the phosphorus atoms, all complexes show little structural differences. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
New Ways to a Series of Parent Representatives of the Eight‐, Nine‐, and Ten‐Vertex Monocarbaborane Family
Authors: Brellochs Bernd , Bačkovský Jaroslav , Štíbr Bohumil , Jelínek Tomáš , Holub Josef , Bakardjiev Mario , Hnyk Drahomír , Hofmann Mathias , Císarová Ivana , Wrackmeyer Berndt
Published: July 2004

The conditions for a reaction betweennido‐B10H14(1) and aqueous formaldehyde in an alkaline solution have been optimized, which resulted in the isolation of the [arachno‐6‐CB9H14]−anion (2) in 52% yield. Anion2was used as a convenient source for the synthesis of a series of basic monocarbaborane compounds. Thus, oxidation of2(Et4N+salt) with I2in CH2Cl2in the presence of Et3N at room temperature afforded the [nido‐6‐CB9H12]−anion (3). Acidification of anion2(generated in situ from borane1) with hydrochloric acid, in the presence of both FeCl3·6H2O and CH2O, led to the formation ofarachno‐4‐CB8H14(4), from whichnido‐1‐CB8H12(5) was obtained by dehydrogenation at 200 °C in quantitative yield. Compound4can easily be converted into the [closo‐1‐CB7H8]−anion (6) in 71% yield via reaction with Et3N and I2in THF at −78 °C. Anion6is also formed in 75% yield by treatment of carborane5with Et3N in refluxing toluene. The [closo‐1‐CB8H9]−anion (7) was isolated by iodination of carborane5in the presence of Et3N (yield 81%) or by the heating of the PPh4+salt of anion2at 220 °C (yield 95%). The structure of anion2was determined by an X‐ray diffraction study and the structures of compounds2,4,5, and7were geometry optimized at the RMP2(fc)/6‐31G* level. The NMR spectroscopic measurements are in some cases complemented by unpublished13C NMR spectroscopic data. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
A New 3‐D Open‐Framework Zinc Phosphate [C6H16N2]·[Zn2(HPO4)3] Synthesized by a Solvothermal Combinatorial Approach
Authors: Song Yu , Yu Jihong , Li Yi , Zhang Min , Xu Ruren
Published: July 2004

By application of a combinatorial approach, a new open‐framework zinc phosphate [C6H16N2]·[Zn2(HPO4)3] (denoted as phase D) has been obtained in the Zn(OAc)2·2H2O/H3PO4/N,N′‐dimethylpiperazine/ethylene glycol solvothermal reaction system at 180 °C. The factors influencing formation of the reaction products, such as crystallization temperature, solvent, as well as gel compositions, have been systematically studied. Phase D was characterized by X‐ray powder diffraction, ICP, and TG analyses, and its structure was determined by single‐crystal X‐ray diffraction analysis. It crystallizes in the monoclinic space group P21/n (no.14), with a = 8.6931(3) Å, b = 14.4394(5) Å, c = 12.6492(3) Å, β = 96.418(2)°, and Z = 4. It is composed of ZnO4, PO3(OH) and PO2(OH)(=O) tetrahedral units linked by Zn−O−P bonds, forming an open‐framework with intersecting eight‐membered ring channels along the [100], [010], and [101] directions. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Behaviour of N‐Pyridylbenzamides versus Benzanilides in the ortho‐Directed Lithiation of Masked Aromatic Carboxylic Acids
Authors: Jóźwiak Andrzej , Brzeziński Jacek Z. , Płotka Mieczysław W. , Szcześniak Aleksandra K. , Malinowski Zbigniew , Epsztajn Jan
Published: July 2004

The reaction of N‐pyridylbenzamides 1−3 with n‐butyllithium or sec‐butyllithium has been examined. The perfect selectivity that has been observed until now in the lithiation of anilides, a reaction used for ortho‐functionalisation of masked aromatic carboxylic acids, has been broken; our results indicate that the pyridine ring at the position ortho to the directed metallation group is more susceptible to lithiation than the homoaromatic ring itself. This was proved in an intermolecular comparative study of benz‐, picolin‐ and isonicotinanilides 14−16, and N‐cumylbenzamide (17). (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Aspects of the Aromaticity of Anthranil
Authors: Matos M. Agostinha R. , Miranda Margarida S. , Morais Victor M. F. , Liebman Joel F.
Published: July 2004

The standard (p° = 0.1 MPa) molar enthalpy of formation of liquid anthranil was measured at T = 298.15 K by static bomb calorimetry and the standard molar enthalpy of vaporization at T = 298.15 K was obtained using Calvet microcalorimetry. These values were used to derive the standard molar enthalpy of formation of anthranil in the gaseous phase. Thermochemical and quantum chemical comparisons were made to interrelate anthranil and its isomers, 1,2‐benzisoxazole, benzoxazole and 2‐cyanophenol, and the monocyclic heterocycles, isoxazole and oxazole. Comparisons with benzofurazan and isobenzofuran were also made. Additionally nucleus‐independent chemical shifts were used as an aromaticity index. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
β‐Imino‐meso‐tetraphenylporphyrin Derivatives in Hetero‐Diels−Alder Reactions
Authors: Alonso Cristina M. A. , Neves Maria G. P. M. S. , Tomé Augusto C. , Silva Artur M. S. , Cavaleiro José A. S.
Published: July 2004

Two types of β‐imino‐meso‐tetraphenylporphyrin derivatives were used as heterodienes in Diels−Alder reactions with electron‐rich dienophiles. These reactions were catalysed by lanthanum triflate. Depending on the starting imine derivative, the products of these reactions are pyrido[2,3‐b]porphyrins or β‐tetrahydroquinoline‐substituted porphyrins. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Generation of a Small Library of Highly Electron‐Rich 2‐(Hetero)Aryl‐Substituted Phenethylamines by the Suzuki−Miyaura Reaction: A Short Synthesis of an Apogalanthamine Analogue
Authors: Appukkuttan Prasad , Orts Amparo Baiget , Chandran R. Prakash , Goeman Jan. L. , Van der Eycken Johan , Dehaen Wim , Van der Eycken Erik
Published: July 2004

The Suzuki−Miyaura reaction is presented as a versatile procedure for the synthesis of a small library of highly electron‐rich 2‐[4,5‐dimethoxy‐2‐(hetero)arylphenyl]ethylamines. Microwave‐irradiation accelerates the reaction tremendously and furnishes superior yields. The difficult oxidative addition of the catalyst to a highly electron‐rich and ortho‐substituted system could be performed easily, and the proto‐deboronation during cross‐coupling reactions involving the highly electron‐withdrawing (2‐formylphenyl)boronic acid could be minimized. Enhanced yields and complete compatibility with aqueous conditions were found. This strategy was developed en route towards the synthesis of an apogalanthamine analogue. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
On the Bis‐Heteronucleophilic Michael Addition to Divinyl Sulfone: A New Efficient Access to Macrocycles by Marie‐Laure Teyssot, Martine Fayolle, Christian Philouze and Claude Dupuy
Authors: Pathak Tanmaya
Published: July 2004

This is a comment on the paper by Dupuy et al. (© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004)
Different mechanisms of chromosome 16 loss of heterozygosity in well‐ versus poorly differentiated ductal breast cancer
Authors: Cleton‐Jansen Anne‐Marie , Buerger Horst , Haar Natalja ter , Philippo Katja , van de Vijver Marc J. , Boecker Werner , Smit Vincent T. H. B. M. , Cornelisse Cees J.
Published: July 2004

Loss of heterozygosity (LOH) at the long arm of chromosome 16 is a frequent genetic alteration in breast cancer. It can occur by physical loss of part of or the entire chromosomal arm, resulting in a decrease in copy number or loss followed by mitotic recombination. Comparative genomic hybridization (CGH) demonstrated that well‐differentiated breast tumors showed significantly more physical loss of 16q than did poorly differentiated ones and that this difference was already discernable in the preinvasive stage. However, polymorphic markers detected no difference in the frequency of 16q LOH between invasive tumors of different histological grade. Here, by combining data on LOH (n = 52), fluorescence in situ hybridization (n = 18) with chromosome 16–specific probes, and CGH (n = 34), we show a preference in well‐differentiated grade I tumors for physical loss of chromosome arm 16q, whereas in poorly differentiated grade III tumors LOH is accompanied by mitotic recombination. This clarifies the discrepancies observed between CGH and LOH for 16q in breast cancer. These different somatic genetic mechanisms may reflect the presence of multiple tumor suppressor genes that are the target of LOH at chromosome arm 16q. © 2004 Wiley‐Liss, Inc.
Co‐localization of differentially expressed genes and shared susceptibility loci in human autoimmunity
Authors: Aune Thomas M. , Parker Joel S. , Maas Kevin , Liu Zheng , Olsen Nancy J. , Moore Jason H.
Published: July 2004

Autoimmune diseases arise from complex interactions between environmental and genetic factors. Genetic linkage scans show that different autoimmune diseases share overlapping susceptibility loci. Lymphocytes from individuals with different autoimmune diseases, as well as unaffected first‐degree relatives, also share a common gene expression profile. We sought to determine if genes within this autoimmune expression profile were nonrandomly distributed in the genome and if their distribution overlapped with shared disease susceptibility loci. We found that differentially expressed genes were distributed in a nonrandom fashion in chromosomal domains within the genome. Furthermore, positions of these domains were not statistically different from a number of shared autoimmune disease susceptibility loci. To our knowledge, this is the first study showing concordance between gene expression and genetic linkage results in common complex multifactorial human diseases. © 2004 Wiley‐Liss, Inc.
Classifying disease chromosomes arising from multiple founders, with application to fine‐scale haplotype mapping
Authors: Yu K. , Martin R.B. , Whittemore A.S.
Published: July 2004

The availability of high‐density haplotype data has motivated several fine‐scale linkage disequilibrium mapping methods for locating disease‐causing mutations. These methods identify loci around which haplotypes of case chromosomes exhibit greater similarity than do those of control chromosomes. A difficulty arising in such mapping is the possibility that case chromosomes have inherited disease‐causing mutations from different ancestral chromosomes (founder heterogeneity). Such heterogeneity dilutes measures of case haplotype similarity. This dilution can be mitigated by separating case chromosomes into subsets according to their putative mutation origin, and searching for an area with excessive haplotype similarity within each subset. We propose a nonparametric method for identifying subsets of case chromosomes likely to share a common ancestral progenitor. By simulation studies and application to published data, we show that the method accurately identifies relatively large subsets of chromosomes that share a common founder. We also show that the method allows more precise estimates of the disease mutation loci than obtained by other fine‐scale mapping methods. © 2004 Wiley‐Liss, Inc.
Assessing genomewide statistical significance in linkage studies
Authors: Lin D.Y. , Zou Fei
Published: July 2004

Assessment of genomewide statistical significance in multipoint linkage analysis is a thorny problem. The existing analytical solutions rely on strong assumptions (i.e., infinitely dense or equally spaced genetic markers that are fully informative and completely observed, and a single type of relative pair) which are rarely satisfied in real human studies, while simulation‐based methods are computationally intensive and may not be applicable to complex data structures and sophisticated genetic models. Here, we propose a conceptually simple and numerically efficient Monte Carlo procedure for determining genomewide significance levels that is applicable to all linkage studies. The pedigree structure is completely general; the marker data are totally arbitrary in respect to number, spacing, informativeness, and missingness; the trait can be qualitative, quantitative, or multivariate; the alternative hypothesis can be two‐sided or one‐sided; and the statistic can be parametric or nonparametric. The usefulness of the proposed approach is demonstrated through extensive simulation studies and an application to the nuclear family data from the Tenth Genetic Analysis Workshop. © 2004 Wiley‐Liss, Inc.
Optimization and dynamics of protein–protein complexes using B‐splines
Authors: Gillilan Richard E. , Lilien Ryan H.
Published: July 2004

A moving‐grid approach for optimization and dynamics of protein–protein complexes is introduced, which utilizes cubic B‐spline interpolation for rapid energy and force evaluation. The method allows for the efficient use of full electrostatic potentials joined smoothly to multipoles at long distance so that multiprotein simulation is possible. Using a recently published benchmark of 58 protein complexes, we examine the performance and quality of the grid approximation, refining cocrystallized complexes to within 0.68 Å RMSD of interface atoms, close to the optimum 0.63 Å produced by the underlying MMFF94 force field. We quantify the theoretical statistical advantage of using minimization in a stochastic search in the case of two rigid bodies, and contrast it with the underlying cost of conjugate gradient minimization using B‐splines. The volumes of conjugate gradient minimization basins of attraction in cocrystallized systems are generally orders of magnitude larger than well volumes based on energy thresholds needed to discriminate native from nonnative states; nonetheless, computational cost is significant. Molecular dynamics using B‐splines is doubly efficient due to the combined advantages of rapid force evaluation and large simulation step sizes. Large basins localized around the native state and other possible binding sites are identifiable during simulations of protein–protein motion. In addition to providing increased modeling detail, B‐splines offer new algorithmic possibilities that should be valuable in refining docking candidates and studying global complex behavior. © 2004 Wiley Periodicals, Inc. J Comput Chem 25: 1630–1646, 2004
Doyen–Wilson theorem for nested Steiner triple systems
Authors: Wang Jinhua , Shen Hao
Published: July 2004

The obvious necessary conditions for the existence of a nested Steiner triple system of order v containing a nested subsystem of order w are v ≥ 3w + 4 and v ≡ w ≡ 1 (mod 6). We show that these conditions are also sufficient. © 2004 Wiley Periodicals, Inc.
Persistent infection of SARS coronavirus in colonic cells in vitro
Authors: Chan Paul K.S. , To Ka‐Fai , Lo Anthony W.I. , Cheung Jo L.K. , Chu Ida , Au Florence W.L. , Tong Joanna H.M. , Tam John S. , Sung Joseph J.Y. , Ng Ho‐Keung
Published: July 2004

Severe acute respiratory syndrome coronavirus (SARS‐CoV) can produce gastrointestinal symptoms. The intestinal tract is the only extrapulmonary site where viable viruses have been detected. This study examined seven established human intestinal cell lines, DLD‐1, HCT‐116, HT‐29, LoVo, LS‐180, SW‐480 and SW‐620, for their permissiveness to SARS‐CoV infection. The results showed that only LoVo cells were permissive to SARS‐CoV infection as evident by positive findings from indirect immunofluorescence staining for intracellular viral antigens, in situ hybridization for intracellular viral RNA, and electron microscopy for intracellular viral particles. In contrast to Vero cells, SARS‐CoV did not produce cytopathic effects on LoVo cells. However, LoVo cells were found to be highly permissive for productive infection with a high viral titre (>3 × 107 viral copies/ml) produced in culture supernatant following a few days of incubation. SARS‐CoV established a stable persistent chronic infection that could be maintained after multiple passages. Being a cell line of human origin, LoVo cells could be a useful in vitro model for studying the biology and persistent infection of SARS‐CoV. Our results on the expression of angiotensin‐converting enzyme 2 (ACE2), a recently identified cellular receptor for SARS‐CoV, in these cell lines indicated that it might not be the sole determinant for cells to be susceptible to SARS‐CoV infection. J. Med. Virol. 74:1–7, 2004. © 2004 Wiley‐Liss, Inc.
Resistance profiles observed in virological failures after 24 weeks of amprenavir/ritonavir containing regimen in protease inhibitor experienced patients
Authors: Marcelin Anne‐Geneviève , Affolabi Dissou , Lamotte Claire , Mohand Hocine Ait , Delaugerre Constance , Wirden Marc , Voujon Delphine , Bossi Philippe , Ktorza Nadine , Bricaire François , Costagliola Dominique , Katlama Christine , Peytavin Gilles , Calvez Vincent ,   the Genophar Study Group
Published: July 2004

Amprenavir (APV) is an HIV protease inhibitor (PI) used for the treatment of either naive or PI‐experienced HIV‐infected patients. Several genotypic resistance pathways in protease gene have been described to be associated to unboosted APV failure (I50V, V32I + I47V, I54L/M, or less commonly I84V, which may be accompanied by one ore more accessory mutations such as L10F, L33F, M46I/L). The aims of this study were to investigate the efficacy up to week 24 of an APV plus ritonavir containing regimen in PI experienced patients and to determine the genotypic resistance profiles emerging in patients failing to this therapy. Forty‐nine, PI experienced but APV naïve patients were treated with APV (600 mg bid) plus ritonavir (100 mg bid). By intent‐to‐treat analysis, the median decrease in viral load (VL) was −1.32 log10 (min +0.6; max −2.8) and −1.46 log10 (min +0.5; max −2.8) 12 and 24 weeks after initiating APV plus ritonavir regimen, respectively. Twelve patients harboured a VL >200 copies/ml at week 24. Among these patients, the selection of mutations previously described with the use of APV as first PI (V32I, L33F, M46I/L, I50V, 54M/L, and I84V) was observed. However, in some cases, mutations classically described after the use of other PIs (V82F and L90M) were selected but always with APV‐specific mutations. There was no relation between the resistance pathways selected with either APV or ritonavir plasma minimal concentration, but higher APV plasma minimal concentration were associated with a lower rate of resistance mutations selection. J. Med. Virol. 74:16–20, 2004. © 2004 Wiley‐Liss, Inc.
Evolution of genotypic and phenotypic resistance to Enfuvirtide in HIV‐infected patients experiencing prolonged virologic failure
Authors: Poveda Eva , Rodés Berta , Labernardière Jean‐Louis , Benito José Miguel , Toro Carlos , González‐Lahoz Juan , Faudon Jean‐Louis , Clavel François , Schapiro Jonathan , Soriano Vincent
Published: July 2004

Four heavily antiretroviral‐experienced HIV‐infected patients had significant plasma HIV‐RNA reductions (>1 log) after beginning an Enfuvirtide (ENF)‐based rescue regimen. However, all had viral rebound shortly thereafter, sustaining high levels of plasma viremia over 80 weeks. These patients developed rapidly genotypic and phenotypic resistance to ENF. Mutations within the HR1 env region were selected (N43D in three and G36V/D in one), resulting in high‐level phenotypic resistance to ENF. Interestingly, two patients had a sustained CD4+ T‐cell increase and two maintained stable CD4+ T‐cell counts despite virologic failure under ENF. The possible mechanisms involved in this response were examined. Changes in virus tropism from R5 to R5/X4 were observed in two patients, in parallel with increases in ENF phenotypic resistance. Low levels of T‐cell activation, T‐cell turnover, and cytotoxic T lymphocyte (CTL) activity were found in all four patients. An overall increase in the proportion of viruses released from cells of the macrophage lineage was observed. In summary, single mutations at the HR1 env region result in significant loss of susceptibility to ENF. Despite virologic failure, these patients may maintain elevated CD4+ counts through a reduction in their overall immune activation. J. Med. Virol. 74:21–28, 2004. © 2004 Wiley‐Liss, Inc.
Consumption of uncooked deer meat as a risk factor for hepatitis E virus infection: An age‐ and sex‐matched case‐control study
Authors: Tei Shuchin , Kitajima Naoto , Ohara Shunji , Inoue Yoshifumi , Miki Masaharu , Yamatani Toshiyuki , Yamabe Hiroshi , Mishiro Shunji , Kinoshita Yoshikazu
Published: July 2004

It was reported previously food‐borne transmission of hepatitis E virus (HEV) to humans from deer meat. The present study attempted to clarify whether eating uncooked deer meat is a major epidemiological risk factor for HEV infection in Kasai, a city in western Japan. In total, 45 volunteer subjects with experience of eating raw deer meat were enrolled. An equivalent number of people from the same area who had never eaten raw deer meat served as controls. The subjects and controls had comparable age and sex distributions. Serum anti‐HEV IgG and anti‐hepatitis A virus (HAV) IgG levels were measured in all 90 volunteers. There was no significant difference in age, overseas travel history, or rate of anti‐HAV antibody positivity between the subjects and controls. Eight (17.7%) of the subjects but only one (2.2%) of the controls had measurable serum anti‐HEV IgG levels (P = 0.014). Anti‐HAV prevalence did not differ between the anti‐HEV‐positive and negative groups. The results suggest that eating uncooked deer meat is an epidemiological risk factor for HEV infection in the studied area. In countries such as Japan where deer meat is sometimes eaten raw, attention must be paid to this route of HEV infection. J. Med. Virol. 74:67–70, 2004. © 2004 Wiley‐Liss, Inc.
Assessment of CMV load in solid organ transplant recipients by pp65 antigenemia and real‐time quantitative DNA PCR assay: Correlation with pp67 RNA detection
Authors: Mengoli Carlo , Cusinato Riccardo , Biasolo Maria A. , Cesaro Simone , Parolin Cristina , Palù Giorgio
Published: July 2004

After bone marrow (BM) or solid‐organ (SO) transplantation viremic Cytomegalovirus (CMV) infection is observed frequently. Quantitative assay of CMV in blood helps the management of this clinical condition. In the present report, 83 samples from 39 solid organ recipients, three CMV assays were compared simultaneously for the first time: the Nuclisens CMV pp67 assay (nucleic acid sequence‐based amplification, NASBA), an “in‐house” quantitative real‐time PCR assay (TaqMan) for CMV DNA, and pp65 antigenemia. The relation between CMV DNA and pp65 antigenemia, the quantitative assays, was evaluated on a larger group including 251 blood samples from 118 solid organ recipients. Real‐time PCR provided the best results; ≥130 CMV DNA copies/2 × 105 peripheral blood leukocytes (PBLs) predicted ≥1 pp65 antigen positive (Ag+) cell/2 × 105 PBLs. By taking pp65 antigenemia as the “gold standard,” the sensitivity of CMV DNA quantitation and of the pp67 RNA assay were 0.95 and 0.20, respectively, while the corresponding specificity values were 0.50 and 0.93. When real‐time PCR was considered as the “gold standard,” the sensitivity and specificity of the pp65 antigenemia were 0.65 and 0.91, respectively. Among the three tests examined, the sensitivity of the pp67 RNA assay was the lowest. On the other hand, the pp67 RNA assay was highly specific and effective in pinpointing high viremia patients. The present report, by providing predictive values for all three diagnostic profiles, DNA load, antigenemia, and pp67RNA, is a contribution for validation of real‐time PCR as a new standard for quantitative assessment of CMV viremia in clinical settings. J. Med. Virol. 74:78–84, 2004. © 2004 Wiley‐Liss, Inc.
Molecular markers of clonality and identity in epstein–barr virus‐associated B‐cell lymphoproliferative disease
Authors: Walling Dennis M. , Andritsos Leslie A. , Etienne Wiguins , Payne Deborah A. , Aronson Judith F. , Flaitz Catherine M. , Nichols C. Mark
Published: July 2004

Epstein–Barr virus (EBV)‐associated B‐cell lymphoproliferative disease may be polyclonal, oligoclonal, or monoclonal. The degree of tumor clonality reflects the disease pathogenesis and may have implications for disease diagnosis, prognosis, and treatment. In this study, specimens of EBV‐associated B‐cell lymphoproliferative disease obtained from immunocompromised hosts were analyzed for molecular markers of cellular and virologic clonality and virologic identity. Each tumor specimen was assessed for immunoglobulin gene JH region rearrangement, the structure of the EBV genome termini, and the EBV genotype(s) present using a new EBV genotyping assay based upon LMP‐1 gene sequence variation. The results of the JH rearrangement and EBV termini assays were generally concordant in their assessment of tumor specimen clonality, and both assays contributed to establishing clonal identity between different tumor specimens. The EBV genotyping assay did not significantly contribute to the assessment of tumor clonality but did established clear virologic identity between different tumor specimens obtained from the same individual. In one individual, these three assays together characterized a multi‐focal, monoclonal tumor that may have arisen through clonal selection after sequential infections with two different EBV genotypes. In summary, the JH rearrangement and EBV termini assays each provided different but complementary information on tumor clonality, while the EBV genotyping assay proved most useful for establishing virologic identity among tumors. Utilization of these three assays together may provide new insight into the pathogenesis of EBV‐associated B‐cell lymphoproliferative disease. J. Med. Virol. 74:94–101, 2004. © 2004 Wiley‐Liss, Inc.
Prevalence and distribution of HSV‐1, VZV, and HHV‐6 in human cranial nerve nuclei III, IV, VI, VII, and XII
Authors: Theil Diethilde , Horn Anja KE , Derfuss Tobias , Strupp Michael , Arbusow Viktor , Brandt Thomas
Published: July 2004

The etiology of idiopathic cranial nerve palsies often remains unresolved. It has been hypothesised that viral reactivation of herpesviruses in the corresponding nuclei in the brainstem is the cause. We investigated the distribution of herpes simplex virus type 1 (HSV‐1) and varicella zoster virus (VZV) in nuclei that are associated with peripheral sensory ganglia [oculomotor (nIII), facial (nVII) nuclei] and in nuclei that are not associated with peripheral sensory ganglia [trochlear (nIV), abducens (nVI), and hypoglossal (nXII) nuclei] of five human brainstems. Samples of the cranial nerve nuclei and adjacent control tissue were taken from histological sections after precise identification of every single nucleus and control tissue. DNA and RNA amplification methods were used to determine the prevalence and distribution of HSV‐1 and VZV. The distribution of human herpes virus type 6 (HHV‐6) was also determined and served as a control, since HHV‐6 infection has never been associated with idiopathic cranial nerve palsies. HSV‐1 was distributed at random in all cranial nerve nuclei and control tissue, whereas VZV DNA was not detected in any of the samples examined. Surprisingly, HHV‐6 was present in almost all samples where HSV‐1 was also present, however, the latency associated transcript (LAT) of HSV‐1 was not found in any of the samples positive for HSV‐1 DNA. The absence of LAT in the samples positive for HSV‐1 and the distribution of HSV‐1 and HHV‐6 do not support the hypothesis that idiopathic cranial nerve palsies result from viral reactivation in the brainstem nuclei. J. Med. Virol. 74:102–106, 2004. © 2004 Wiley‐Liss, Inc.
New point of care test is highly specific but less sensitive for influenza virus A and B in children and adults
Authors: Rawlinson William D. , Waliuzzaman Zubair M. , Fennell Michael , Appleman James R. , Shimasaki Craig D. , Carter Ian W.
Published: July 2004

The importance of rapid diagnosis of influenza has increased with the availability of neuraminidase inhibitors, which need to be commenced within 48 hr of symptom onset. Furthermore, the recent development of influenza‐like clinical syndromes with novel aetiologies (severe acute respiratory syndrome, SARS) has increased the need for rapid and accurate near‐patient diagnosis. A new, modified point of care (POC) diagnostic test (ZstatFlu) was assessed on 469 nasopharyngeal aspirates (NPAs) and 260 nose/throat swabs (TS) taken from children and adults. The test was specific (77–98%) for all specimen types for influenza virus A and B, depending upon incubation conditions. However, it was less sensitive, detecting 65–77% of specimens confirmed as positive on culture, direct immunofluorescence or PCR testing. A positive test is useful, for both directing initiation of therapy in the clinician's office, and making a positive diagnosis of influenza in patients with influenza‐like clinical syndromes. J. Med. Virol. 74:127–131, 2004. © 2004 Wiley‐Liss, Inc.
Genetic diversity of influenza B virus: The frequent reassortment and cocirculation of the genetically distinct reassortant viruses in a community
Authors: Matsuzaki Yoko , Sugawara Kanetsu , Takashita Emi , Muraki Yasushi , Hongo Seiji , Katsushima Noriko , Mizuta Katsumi , Nishimura Hidekazu
Published: July 2004

To characterize the genetic diversity of influenza B viruses isolated during one influenza season, the antigenic and genetic relationships among 20 strains of influenza B virus isolated in February and March 2001 at one pediatric clinic in Yamagata City, Japan, were investigated. The HA gene and seven other gene segments were phylogenetically divided into three distinct sublineages (Harbin/7/94‐, Tokyo/6/98‐, and Shiga/T30/98‐related lineage) of the Yamagata/16/88‐like lineage. The NS genes of the viruses belonging to the Harbin/7/94‐related lineage have additional three nucleotides at positions 439–447, and were phylogenetically distinguishable from those of the currently circulating Yamagata/16/88‐ and Victoria/2/87‐like lineages, but were closely related to that of the Yamagata/16/88‐like lineage isolated before 1994. Moreover, four strains of influenza B virus isolated in the same community between 2002 and 2003 were further examined. Phylogenetic analysis revealed that a virus of Victoria/2/87‐like lineage isolated in 2003 had acquired the NA, NS, M, and PA gene segments from a Shiga/T30/98‐like virus, and two strains of Harbin/7/94‐related lineage had acquired the various gene segments from Shiga/T30/98‐like virus through a reassortment event. These results indicate that genetically distinct multiple viruses can combine to cause an influenza B epidemic in a community and that the frequent reassortment among these viruses plays a role in generating the genetic diversity of influenza B viruses. J. Med. Virol. 74:132–140, 2004. © 2004 Wiley‐Liss, Inc.