bluuebunny/crossref_metadata_2025 huggingface dataset:
Synthesis of Novel Functionalized Symmetric Bi‐2(1H)‐pyrazinones.
Authors: De Borggraeve Wim M. , Appukkuttan Prasad , Azzam Rasha , Dehaen Wim , Compernolle Frans , Van der Eycken Erik , Hoornaert Georges
Published: July 2005

For Abstract see ChemInform Abstract in Full Text.
Synthesis of a Selenium‐Substituted Diselenide.
Authors: Cox Matthew , Wirth Thomas
Published: July 2005

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Selenium‐Containing π‐Conjugated Compounds for Electronic Molecular Materials.
Authors: Takimiya Kazuo , Otsubo Tetsuo
Published: July 2005

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Carbonylation of Lithium Enolates of Esters and Amides with Carbon Monoxide and Selenium
Authors: Kambe Nobuaki , Nishiyama Akira , Fujiwara Shin‐Ichi , Tsutomu Shin‐Ike , Sonoda Noboru
Published: July 2005

For Abstract see ChemInform Abstract in Full Text.
Concise Synthesis of (.+‐.)‐Smenochromene D (= Likonide B).
Authors: Olson Brooke S. , Trauner Dirk
Published: July 2005

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Cytotoxic Iridoid Aldehydes from Taiwanese Viburnum luzonicum.
Authors: Fukuyama Yoshiyasu , Minoshima Yuka , Kishimoto Yoshiko , Chen Ih‐Sheng , Takahashi Hironobu , Esumi Tomoyuki
Published: July 2005

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New Cassane‐Type Diterpenes of Caesalpinia crista from Myanmar.
Authors: Kalauni Surya Kant , Awale Suresh , Tezuka Yasuhiro , Banskota Arjun Hari , Linn Thein Zaw , Kadota Shigetoshi
Published: July 2005

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Enantioselective Annulation Using Nazarov Reagent: Synthesis of (+)‐Preoleanatetraene.
Authors: Barrero Alejandro F. , Arseniyadis Simeon , Quilez del Moral Jose F. , Mar Herrador M. , Rosellon Antonio
Published: July 2005

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Diastereoselective Synthesis of (S)‐ and (R)‐α‐Phenylserine by a Sulfinimine‐Mediated Strecker Reaction.
Authors: Avenoza Alberto , Busto Jesus H. , Corzana Francisco , Peregrina Jesus M. , Sucunza David , Zurbano Maria M.
Published: July 2005

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Efficient and Selective Deuteration of Phenylalanine Derivatives Catalyzed by Pd/C.
Authors: Maegawa Tomohiro , Akashi Akira , Esaki Hiroyoshi , Aoki Fumiyo , Sajiki Hironao , Hirota Kosaku
Published: July 2005

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Synthesis of Novel Anellated Pyranosides as Precursors of C‐Nucleoside Analogues Using Isopropyl 6‐O‐Acetyl‐3‐deoxy‐4‐S‐ethyl‐4‐thio‐α‐D‐threo‐hexopyranosid‐2‐ulos e.
Authors: Ruiz Reinaldo Molina , Martinez Iran Otero , Michalik Manfred , Reinke Helmut , Suarez Jose Quincoces , Peseke Klaus
Published: July 2005

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Synthesis and Stereoselective Glycosylation of D‐ and L‐glycero‐β‐D‐manno‐Heptopyranoses.
Authors: Crich David , Banerjee Abhisek
Published: July 2005

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An Aza‐Wittig/π‐Furan Cyclization Approach Toward the Homoerythrina Alkaloid (.+‐.)‐Selaginoidine.
Authors: Cassidy Michael P. , Oezdemir Ayse Daut , Padwa Albert
Published: July 2005

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Stereoselective Synthesis of the Octahydronaphthalene Unit of Integramycin (I) via an Intramolecular Diels—Alder Reaction.
Authors: Dineen Thomas A. , Roush William R.
Published: July 2005

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A New and Known Cytotoxic Aryltetralin‐Type Lignans from Stems of Bursera graveolens.
Authors: Nakanishi Tsutomu , et al. et al.
Published: July 2005

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A Short, Stereoselective, and Common Approach for the Synthesis of 4,5‐Disubstituted δ‐Lactones Simplactone B and Its C‐5 Analogue.
Authors: Kumar A. Ravi , Sudhakar N. , Rao B. Venkateswara , Raghunandan N. , Venkatesh A. , Sarangapani M.
Published: July 2005

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High Density Hydroxyl Anions in a Microporous Crystal: [Ca24Al28O64] 4+×4(OH‐).
Authors: Li Jiang , Huang Fan , Wang Lian , Yu Shu‐Qin , Torimoto Youshifumi , Sadakata Masayoshi , Li Quan Xin
Published: July 2005

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Apparent Two‐Dimensional Behavior of TiO2 Nanotubes Revealed by Light Absorption and Luminescence.
Authors: Bavykin Dmitry V. , Gordeev Sergey N. , Moskalenko Andriy V. , Lapkin Alexei A. , Walsh Frank C.
Published: July 2005

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Microwave Approach for the Synthesis of Rhabdophane‐Type Lanthanide Orthophosphate (Ln: La, Ce, Nd, Sm, Eu, Gd and Tb) Nanorods under Solvothermal Conditions.
Authors: Patra Chitta Ranjan , Alexandra Gabashvili , Patra Sujata , Jacob David Solomon , Gedanken Aharon , Landau Asher , Gofer Yossi
Published: July 2005

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Synthesis and Pharmacological Activity of 4‐(4′‐(Chlorophenyl)‐4‐hydroxypiperidine) Derivatives.
Authors: Saify Zafar Saeed , Mushtaq Nousheen , Khan Khalid Mohammed , Perveen Shahnaz , Shah Syed Tasadaque Ali , Abdel‐Jalil Raid Jamil , Fecker Miriam , Voelter Wolfgang
Published: July 2005

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Discovery of a Novel and Potent Human and Rat β3‐Adrenergic Receptor Agonist, [3‐[(2R)‐[ [(2R)‐(3‐Chlorophenyl)‐2‐hydroxyethyl]amino]propyl]‐1H‐indol‐7‐yloxy] acetic Acid.
Authors: Harada Hiroshi , Hirokawa Yoshimi , Suzuki Kenji , Hiyama Yoichi , Oue Mayumi , Kawashima Hitoshi , Kato Hiroshi , Yoshida Naoyuki , Furutani Yasuji , Kato Shiro
Published: July 2005

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Kondo Effect on Mesoscopic Scale
Authors: Ujsaghy O. , Zawadowski A.
Published: July 2005

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Azobenzene as Photoresponsive Conformational Switch in Cyclic Peptides
Authors: Renner C. , Kusebauch U. , Loeweneck M. , Milbradt A. G. , Moroder L.
Published: July 2005

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Developments in Catalytic Aromatic C—H Transformations — Promising Tools for Organic Synthesis
Authors: Goj Laurel A. , Gunnoe T. Brent
Published: July 2005

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Environmentally Friendly Suzuki Aryl—Aryl Cross‐Coupling Reaction
Authors: Bai L. , Wang J.‐X.
Published: July 2005

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Rhodium(I)‐Catalyzed [4 + 2] and [4 + 2 + 2 ] Carbocyclizations
Authors: Robinson John E.
Published: July 2005

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Rhodium(I)‐Catalyzed [2 + 2 + 1] and [4 + 1] Carbocyclization Reactions
Authors: Jeong Nakcheol
Published: July 2005

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Rhodium(I)‐Catalyzed Cycloisomerization and Cyclotrimerization Reactions
Authors: Fujiwara Masaki , Ojima Iwao
Published: July 2005

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Synergic Photosensitizers: A New Trend in Photodynamic Therapy
Authors: Lunardi Claure N. , Tedesco Antonio C.
Published: July 2005

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Rearrangement Processes of Oxonium and Ammonium Ylides Formed by Rhodium(II)‐Catalyzed Carbene Transfer
Authors: West Frederick F.
Published: July 2005

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Rhodium(II)‐Catalyzed Oxidative Amination
Authors: Espino Christine G. , Du Bois Justin
Published: July 2005

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Magnesium Oxide, Alkoxides, and Carboxylates
Authors: Shimizu M.
Published: July 2005

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Magnesium—Metal Reagents
Authors: Zhang J.‐H. , Keh C. C. K. , Li C.‐J.
Published: July 2005

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Diazo Compounds
Authors: Heydt H.
Published: July 2005

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Preparation of Kinase‐Biased Compounds in the Search for Lead Inhibitors of Kinase Targets
Authors: Langston Steven , et al. et al.
Published: July 2005

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A Decade of Advances in Three‐Component Reactions
Authors: Syamala M.
Published: July 2005

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Synthesis of Orally Active Small‐Molecule Gonadotropin‐Releasing Hormone Antagonists
Authors: Tucci Fabio C. , Chen Chen
Published: July 2005

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Rhodium(II)‐Stabilized Carbenoids Containing Both Donor and Acceptor Substituents
Authors: Davies Huw M. L. , Walji Abbas M.
Published: July 2005

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Rhodium(I)‐Catalyzed Allylic Substitution Reactions and Their Applications to Target Directed Synthesis
Authors: Leahy David K. , Evans P. Andrew
Published: July 2005

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Carbon—Carbon Bond‐Forming Reactions Starting from Rh—H or Rh—Si Species
Authors: Matsuda Isamu
Published: July 2005

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Stereoselective Rhodium(I)‐Catalyzed Hydroformylation and Silylformylation Reactions and Their Application to Organic Synthesis
Authors: Leighton James L.
Published: July 2005

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Gas‐Phase Radical Chemistry in the Troposphere
Authors: Monks Paul S.
Published: July 2005

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Rhodium‐Catalyzed Asymmetric Hydrogenation
Authors: Chi Yongxiang , Tang Wenjun , Zhang Xumu
Published: July 2005

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Biomimetic Assemblies of Carbon Nanostructures for Photochemical Energy Conversion
Authors: Guldi Dirk M.
Published: July 2005

For Abstract see ChemInform Abstract in Full Text.
Gene expression of the repulsive guidance molecules during development of the mouse intestine
Authors: Metzger Marco , Conrad Sabine , Alvarez‐Bolado Gonzalo , Skutella Thomas , Just Lothar
Published: July 2005

Repulsive guidance molecules (RGMs) are recently identified proteins implicated in neuronal differentiation, migration, and apoptosis. However, in non‐neural tissues a specific biological function of RGM is still unknown. In this study, we describe the expression patterns of the RGM members (a, b, and c) during embryonic and postnatal development of the small and large murine intestine. We demonstrated by RT‐PCR, in situ hybridization, Western blot, and immunocytochemistry that subtypes RGMa and RGMb but not RGMc were strongly expressed in enteric ganglia cells of the fetal and adult gut. In contrast to the enteric nervous system, RGMa and RGMb expression in the intestinal epithelium started during postnatal gut development. Interestingly, both subtypes were predominantly expressed in the proliferative crypt compartment of the gut epithelium and in paneth cells of small intestine. The development‐dependent expression in enteric ganglia and intestinal epithelial cells suggests that RGM may be involved in cell migration, differentiation, and apoptosis with similar cellular mechanisms as described in the central nervous system. Developmental Dynamics 234:169–175, 2005. © 2005 Wiley‐Liss, Inc.
Prevalence of 9p21 deletions in UK melanoma families
Authors: Mistry Sushila H. , Taylor Claire , Randerson‐Moor Juliette A. , Harland Mark , Turner Faye , Barrett Jennifer H. , Whitaker Linda , Jenkins Robert B. , Knowles Margaret A. , Bishop Julia A. Newton , Bishop D. Timothy
Published: July 2005

Although the CDKN2A gene has been shown to be the major genetic determinant governing high‐penetrance susceptibility to melanoma, there remains a significant proportion of melanoma pedigrees in which germline mutations of CDKN2A have not been identified. We have therefore studied the prevalence of germline 9p deletions encompassing the CDKN2 locus in melanoma pedigrees, using multiplex ligation‐dependent probe amplification. Germline deletions were found in 3 of 93 UK pedigrees, with no previously identified CDKN2A mutations. A hemizygous deletion of CDKN2A exon 1β previously reported by this group was confirmed in one family and identified in a second. Microsatellite analysis determined that these two families were ancestrally related. In the third family, a novel p16 hemizygous deletion involving CDKN2A exons 1α, 2, and 3 was detected. An additional 9p21 deletion reported previously in a USA melanoma‐neural system tumor family was shown to involve CDKN2A exon 1β, and not p16. The CDKN2A exon 1β deletions provide further evidence that this tumor suppressor gene is important in melanoma‐neural system tumor susceptibility, but do not exclude the possibility of a novel gene or regulatory element also being deleted in this region. Deletions at 9p21 are rare and explain only a small proportion of melanoma susceptibility. This study is the first to comprehensively exclude deletions in melanoma‐prone families with no previously identified CDKN2A mutations. © 2005 Wiley‐Liss, Inc.
Coffee consumption and breast cancer risk among BRCA1 and BRCA2 mutation carriers
Authors: Nkondjock André , Ghadirian Parviz , Kotsopoulos Joanne , Lubinski Jan , Lynch Henry , Kim‐Sing Charmaine , Horsman Douglas , Rosen Barry , Isaacs Claudine , Weber Barbara , Foulkes William , Ainsworth Peter , Tung Nadine , Eisen Andrea , Friedman Eitan , Eng Charis , Sun Ping , Narod Steven A.
Published: July 2005

Although there are several plausible biologic mechanisms whereby coffee consumption might influence the risk of breast cancer, epidemiologic evidence is limited. We assessed the association between coffee consumption and breast cancer risk among high‐risk women who carry BRCA mutations. We performed a matched case‐control analysis on 1,690 women with a BRCA1 or BRCA2 mutation from 40 centers in 4 countries. Average lifetime coffee consumption was estimated via a self‐administered questionnaire. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using conditional logistic regression. After adjustment for potential confounders, the ORs for breast cancer in BRCA carriers who habitually drank 0, 1–3, 4–5 and 6 or more cups of coffee were 1.00, 0.90 (95% CI 0.72–1.12), 0.75 (95% CI 0.47–1.19) and 0.31 (95% CI 0.13–0.71; p‐trend = 0.02). The effect was limited to the consumption of caffeinated coffee. These results suggest that among women with BRCA gene mutation, coffee consumption is unlikely to be harmful and that high levels of consumption may in fact be related to reduced breast cancer risk. © 2005 Wiley‐Liss, Inc.
An inverse association between ovarian cysts and breast cancer in the breast cancer family registry
Authors: Knight Julia A. , John Esther M. , Milne Roger L. , Dite Gillian S. , Balbuena Ron , Shi Ellen J.Q. , Giles Graham G. , Ziogas Argyrios , Andrulis Irene L. , Whittemore Alice S. , Hopper John L. ,   for the Breast Cancer Family Registry
Published: July 2005

Ovarian cysts of several types are common in women of reproductive age. Their etiology is not well understood but is likely related to perturbations in the hypothalamic‐pituitary‐gonadal axis. The relationship of ovarian cysts to breast cancer risk is not known, although a negative association with polycystic ovarian syndrome has been reported. Incident, invasive female breast cancer cases, population‐based controls and unaffected sisters of cases were studied from 3 countries participating in the Breast Cancer Family Registry: Melbourne and Sydney, Australia; the San Francisco Bay Area, USA; and Ontario, Canada. Using the same questionnaire, information was collected on self‐reported history of ovarian cysts and other risk factors. Analyses were based on 3,049 cases, 2,344 population controls and 1,934 sister controls from all sites combined. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using both unconditional and conditional logistic regression using an offset term to account for sampling fractions at 2 of the sites. A significantly reduced risk of breast cancer was observed for women reporting a history of ovarian cysts (OR = 0.70, 95% CI 0.59–0.82, among all cases and all controls). This risk estimate was similar regardless of control group used, within all 3 sites and in both premenopausal and postmenopausal women (ORs ranging from 0.68–0.75, all 95% CI excluded 1.00). A self‐reported history of ovarian cysts was strongly and consistently associated with a reduced risk of breast cancer. Further study of ovarian cysts may increase our understanding of hormonal and other mechanisms of breast cancer etiology. © 2005 Wiley‐Liss, Inc.
Biases in human papillomavirus genotype prevalence assessment associatedwith commonly used consensus primers
Authors: Chan Paul K. S. , Cheung Tak‐Hong , Tam Ann O. Y. , Lo Keith W. K. , Yim So‐Fan , Yu May M. Y. , To Ka‐Fai , Wong Yick‐Fu , Cheung Jo L. K. , Chan Denise P. C. , Hui Mamie , Ip Margaret
Published: July 2005

Consensus primers targeting human papillomaviruses (HPVs) have biases in sensitivity toward certain HPV types. We applied 3 primer sets (GP5+/6+, MY09/11, PGMY09/11) in parallel on 120 Chinese cervical cancer specimens. GP5+/6+ exhibited a poor sensitivity for HPV52, for which the prevalence among squamous cell cervical cancer was underestimated from 14.6% to 0%. The fact that HPV52 should rank second in prevalence among squamous cell cervical carcinoma in Hong Kong could be missed if GP5+/6+, a worldwide commonly used primer set, was selected for HPV detection. Biases in HPV type‐specific sensitivity may result in misprioritization of vaccine candidates. © 2005 Wiley‐Liss, Inc.
Testicular germ cell tumors exhibit evidence of hormone dependence
Authors: Douglas Meaghan L. , Richardson Michelle M. , Nicol David L.
Published: July 2005

The aim of this investigation was to test the hypothesis that testicular germ cell tumors (TGCTs) are hormone‐dependent cancers. Human TGCT cells were implanted in the left testis of male severe combined immunodeficient mice receiving either no treatment or hormone manipulation treatment [blockade of gonadotropin‐releasing hormone secretion and/or signaling using leuprolide or leuprolide plus exogenous testosterone]. Real‐time RT‐PCR analysis was used to determine the expression profiles of hormone pathway‐associated genes. Tumor burden was significantly smaller in mice receiving both leuprolide and testosterone. Real‐time RT‐PCR analysis of follicle‐stimulating hormone (FSH) receptor, luteinizing hormone (LH) receptor and P450 aromatase revealed changes in expression in normal testis tissue related to presence of xenograft tumors and manipulation of hormone levels but a complete absence of expression of these genes in tumor cells themselves. This was confirmed in human specimens of TGCT. Reduced TGCT growth in vivo was associated with significant downregulation of LH receptor and P450 aromatase expression in normal testes. In conclusion, manipulation of hormone levels influenced the growth of TGCT in vivo, while the presence of xenografted tumors influenced the expression of hormone‐related genes in otherwise untreated animals. Human TGCTs, both in the animal model and in clinical specimens, appear not to express receptors for FSH or LH. Similarly, expression of the P450 aromatase gene is absent in TGCTs. Impaired estrogen synthesis and/or signaling may be at least partly responsible for inhibition of TGCT growth in the animal model. © 2005 Wiley‐Liss, Inc.
Association between serum pepsinogens and polymorphismof PTPN11 encoding SHP‐2 among Helicobacter pylori seropositive Japanese
Authors: Goto Yasuyuki , Ando Takafumi , Yamamoto Kazuhito , Tamakoshi Akiko , El‐Omar Emad , Goto Hidemi , Hamajima Nobuyuki
Published: July 2005

Helicobacter pylori (H. pylori) plays a crucial role in the development of gastric atrophy and cancer, and cagA‐positive strains, which are universal in Japan, increase the risk of these outcomes substantially. The CagA protein is injected from attached H. pylori into gastric epithelial cells and undergoes Src‐dependent tyrosine phosphorylation and activation of the eukaryotic phosphatase SHP‐2. The CagA/SHP‐2 interactions elicit cellular changes that increase the risk of carcinogenesis. We investigated the association of a frequent single nucleotide polymorphism (SNP; JST057927; G‐to‐A) in the PTPN11 gene that encodes SHP‐2 with gastric atrophy and gastric cancer in Japan. Gastric atrophy was assessed by measuring serum pepsinogen I and II levels. The subjects comprised 454 healthy controls (126 males; mean age, 58.4) and 202 gastric cancer cases (134 males and 68 females; mean age, 66.7). All gastric cancer cases and 250 (55%) controls were H. pylori seropositive; 179 (89%) of the gastric cancer cases had gastric atrophy compared to 137 (55%) of the H. pylori seropositive controls (p < 0.001). Among HP seropositive controls compared to the common PTPN11 G/G genotype, the odds ratio of atrophy was nonsignificantly reduced with the G/A genotype (0.70; 95% CI = 0.39–1.25) and significantly reduced with the A/A genotype (0.09; 95% CI = 0.01–0.72). Lower risk for gastric atrophy had a gene‐dose association with the A allele (p = 0.01, trend test). There was a clear deficiency of the A/A genotype in those with atrophy compared to those without (1 subject in the gastric atrophy group vs. 8 in the group without). Cancer cases differed from controls in frequencies of PTPN11 G/A genotype only because of a higher prevalence of atrophy among the cancer cases. The G/A SNP in the PTPN11 gene appears to be a risk factor for gastric atrophy in subjects infected with cagA‐positive H. pylori. This may explain why only a proportion of CagA‐positive individuals develop gastric atrophy and gastric cancer, even though infection with cagA strains is universal in Asian countries such as Japan. The functional consequences of the G/A polymorphism remain to be elucidated. © 2005 Wiley‐Liss, Inc.
Application research of plasma‐enhanced electrochemical surface ceramic‐coating technology on titanium implants
Authors: Nan Kaihui , Wang Yingjun , Chen Xiaofeng , Ning Chengyun , Wang lingyu , Zhao Naru
Published: July 2005

Plasma electrochemical surface ceramic coating (PECC) is an enhanced electrochemistry technique accomplished by means of plasma arc discharge in an electrolyte medium obtain coating structures on valve‐metal surfaces. Oxide films of titanium obtained by PECC in the electrolyte of sodium phosphate were investigated. The film was composed mainly of TiO2 in the form of anatase and rutile and enriched with Na and P elements on their surfaces. Their apatite‐inducing ability was also evaluated in a simulated body fluid (SBF). When immersed in SBF for over 30 days, an oriented hydroxy‐carbonate‐apatite was induced on the surface of the films, which suggests the PECC‐treated titanium has the promising positive biological response. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2005
Initial adhesion of glass‐fiber–reinforced composite to the surface of porcine calvarial bone
Authors: Tuusa S. M. R. , Lassila L. V. J. , Matinlinna J. P. , Peltola M. J. , Vallittu P. K.
Published: July 2005

The aim of this preliminary study was to compare the initial bond strength of the glass‐fiber–reinforced composite veil to the surface of the porcine calvarial compact bone using different adhesives. Fiber‐reinforced composite (FRC) made of E‐glass fiber veil with the BisGMA‐PMMA resin system was used in the study. For the shear bond strength test, porcine calvarial bone cubes were mounted into resin matrix. FRC‐veil discs were bonded to compact bone with different types of adhesives: (A) BisGMA‐HEMA based (3M‐ESPE Scotchbond Multi‐Purpose Adhesive), (B) 4‐META/UDMA/BisGMA based (Unifil Bond Bonding Agent) and MDP based (Clearfil Se Bond adhesive), (C) UDMA/BisGMA/PMMA‐based experimental adhesive, and (D) silane‐based (APS, ICS, MPS) experimental adhesives. The surface of the bone was mechanically roughened and was either used as such, treated with dental primers (Unifil Bond Self‐etching Primer, Clearfil Se Bond Primer), or treated with an experimental silane mixture (APS, ICS, MPS), or with a mixture of the experimental silane liquid and Clearfil Se Bond Primer. The 3M‐ESPE Scotchbond Multi‐Purpose Adhesive and UDMA/BisGMA/PMMA experimental adhesive gave poor results in the shear bond test (0.58 and 0.40 MPa, respectively). Unifil Bond Bonding Agent and Clearfil Se Bond adhesive with respective primers markedly improved the shear bond strength; with Unifil the result was 3.40 MPa, and with Clearfil it was 6.19 MPa. When the bone surface was primed with a mixture of Clearfil Se Bond Primer and Clearfil Porcelain Bond Activator, the Clearfil Se Bond adhesive‐impregnated FRC veil gave the best adhesion to the bone surface in this test: 9.50 MPa. The addition of bioactive glass granules between the veil and the bone lowered the shear bond strength in the test system described above to 6.72 MPa. The test systems with the silane mixture were also promising. In the SEM study, it was found that the mechanical treatment reveals the pores of the bone surface. Chemical treatments of the bone surface improved the adhesion of the FRC veil to the bone. The results showed that the adhesion of the FRC to the surface of the bone can be significantly improved with mechanical roughening and with special chemical treatments of the bone surface. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2005
In vitro evaluation of caries inhibition promoted by self‐etching adhesive systems containing antibacterial agents
Authors: Lobo Maristela M. , Gonçalves Reginaldo B. , Pimenta Luiz André F. , Bedran‐Russo Ana Karina B. , Pereira Patrícia N. R.
Published: July 2005

This study evaluated the cariostatic effect of antibacterial self‐etching adhesive systems, by means of an in vitro bacterial caries model. Seventy‐five prepared bovine slabs were randomly divided into groups (n = 15): (1) unbonded composite, no carious challenge (UNB‐NC); (2) unbonded composite, carious challenge (UNB‐C); (3) Clearfil SE Bond, no antibacterial agent (CSE); (4) Protect Bond, containing MDPB and fluoride (PB); and (5) Reactmer Bond, fluoride‐releasing (RB). All preparations were restored with Filtek™ Z‐250. Groups (2)–(5) were submitted to a medium containing Streptococcus mutans (ATCC ‐ 25175) for 5 days, and Group (1) was kept in a noninoculated medium. Insoluble polysaccharides present in tooth biofilms were quantified, Knoop hardness (KHN) was measured on the enamel adjacent to restorations, and standard 35‐mm polarized light photomicrographs were taken as illustrations. Polysaccharide and Knoop hardness results were analyzed with the use of ANOVA, with a split‐split–plot statistical design for KHN. Except for Group (1), all groups showed similar caries formation. Biofilm over PB restorations showed the smallest amounts of polysaccharides (14.37 μg/mg), and CSE showed the highest amounts (20.87 μg/mg). All self‐etching systems tested were unable to inhibit secondary caries in a bacterial model simulating a high caries challenge, even though there was reduced glucan synthesis provided by the adhesive system containing MDPB and fluoride. © 2005 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2005
Serum albumin enhances polyethylenimine‐mediated gene delivery to human respiratory epithelial cells
Authors: Carrabino Salvatore , Di Gioia Sante , Copreni Elena , Conese Massimo
Published: July 2005

Background

The interaction of polyethylenimine (PEI) polyplexes with proteins in cystic fibrosis (CF) airway secretions poses a significant hurdle to this nonviral delivery system. The aim of this study was to evaluate whether albumin may increase the efficiency of PEI complexes in mediating gene transfer into respiratory epithelial cells in the presence of CF mucus.

Methods

PEI (25 kDa) was complexed to DNA in the presence of human serum albumin (HSA) and used to transfect confluent A549 and 9HTEo‐ cells. Alternatively, albumin was added to preformed PEI‐DNA complexes. The cytotoxicity of complexes was analysed by the LDH (lactate dehydrogenase) assay. CF CFT1‐C2 cells were allowed to polarise and were transfected either with luciferase‐ or CFTR‐expressing plasmids. To evaluate the effect of CF respiratory secretions on transfection efficiency, confluent cells were transfected in the presence of sputum obtained from two CF patients.

Results

The ternary PEI‐HSA complexes increased luciferase expression in confluent cultures in a dose‐dependent fashion up to 100 times as compared to PEI‐DNA. The number of GFP‐expressing cells, as evaluated by epifluorescence, was augmented several‐fold. When HSA was added to preformed PEI‐DNA complexes, a further 5–10‐fold increase in gene expression was observed. No significant cytotoxicity was observed with either PEI or PEI‐HSA polyplexes. The ternary complexes determined detectable CFTR gene transfer and expression at the apical membrane in polarised CFT1‐C2 cells, as evaluated by confocal microscopy. CF sputum inhibited PEI‐mediated gene transfer by 7–186‐fold. Although luciferase expression mediated by PEI‐HSA was still inhibited by CF sputum, these levels were 18–83.8‐fold higher than with PEI.

Conclusions

Our results demonstrate that albumin increases PEI gene transfer efficiency in confluent and polarised respiratory epithelial cells and can allow CFTR gene expression in the appropriate cellular compartment. PEI‐HSA complexes display a higher efficiency than PEI also in the presence of CF sputum, indicating that albumin‐containing polyplexes may help overcome barriers imposed by CF airway secretions. Copyright © 2005 John Wiley & Sons, Ltd.
Impact of drug resistance genotypes on CD4+ counts and plasma viremia in heavily antiretroviral‐experienced HIV‐infected patients
Authors: Rodes Berta , García Federico , Gutierrez Carolina , Martinez‐Picado Javier , Aguilera Antonio , Saumoy Maria , Vallejo Alex , Domingo Pere , Dalmau David , Ribas Maria Angels , Blanco José Luis , Pedreira José , Perez‐Elias Maria Jesús , Leal Manuel , de Mendoza Carmen , Soriano Vincent
Published: July 2005

The number of HIV‐infected individuals with prior multiple treatment failures is increasing as time passes by. The success of antiretroviral therapy in these patients is often compromised by the selection of drug‐resistant viruses. Despite initial concerns, a rebound in AIDS cases among heavily treatment‐experienced patients failing virologically their antiretroviral therapy has not occurred yet. In a multicenter study conducted in Spain, HIV‐infected patients were assessed with prior failure to antiretrovirals from the three main drug families who presented during the last semester of the year 2003 with plasma HIV‐RNA values above 1,000 copies/ml, despite good treatment adherence. The relationships between CD4+ T cell counts, viral loads and drug‐resistant genotypes were examined. A total of 273 patients were identified in 12 centers (78% male, median age: 41 years). The mean viral load was 50,438 copies/ml and the mean CD4+ count was 328 cells/μl. Only 19.5% had less than 200 CD4+ T cells/μl. Most patients (95%) were receiving nucleoside reverse transcriptase inhibitors (NRTI) in their last antiretroviral regimen, while 63% were treated with protease inhibitors (PI) and 27% on non‐nucleoside reverse transcriptase inhibitors (NNRTI). Overall, 97.4% had at least one drug resistance mutation (87.2% for NRTI, 68.5% for NNRTI, and 92.7% for PI). Using the virtual phenotype, resistance to three or more drugs within each class was recognized in 45.8% for NRTI, 40.7% for NNRTI, and 44.7% for PI. Moreover, cross‐resistance to compounds from two or three drug families was recognized in 41% and 19.4% of patients, respectively. Nearly half of the patients had plasma HIV‐RNA below 10,000 copies/ml and they showed significantly higher CD4 + counts than those with greater viremia (408 versus 259 cells/μl; P < 0.001). Patients with higher plasma viremia had significantly more drug resistance mutations than those with lower viremia. No favorable effect on viral load could be recognized for individual drug resistance mutations known to reduce viral fitness in vitro (i.e., rtM184V, rtL74V, rtK65R, proD30N, or proI50L). In summary, a large proportion of treatment‐experienced patients failing their current antiretroviral regimen carry viruses with broad cross‐resistant genotypes. Nearly half of the patients with these multi‐drug resistant viruses had < 10,000 HIV‐RNA copies/ml and 80% have more than 200 CD4 + T cells/μl. Thus, maintaining treatment HIV‐infected individuals failing virologically and harboring drug‐resistant viruses might ameliorate immunological deterioration until new drugs became available. J. Med. Virol. 77:23–28, 2005. © 2005 Wiley‐Liss, Inc.
Autoantibodies against human epithelial cells and endothelial cells after severe acute respiratory syndrome (SARS)‐associated coronavirus infection
Authors: Yang Yao‐Hsu , Huang Yu‐Hui , Chuang Ya‐Hui , Peng Chung‐Min , Wang Li‐Chieh , Lin Yu‐Tsan , Chiang Bor‐Luen
Published: July 2005

The severe acute respiratory syndrome (SARS) is caused by infection with the SARS‐associated coronavirus (SARS‐CoV) and characterized by severe pulmonary inflammation and fibrosis. In this study, the development of autoantibodies against human epithelial cells and endothelial cells in patients with SARS at different time periods (the first week: phase I, 1 month after the disease onset: phase II/phase III) were investigated. Antibodies in sera of patients and healthy controls against: (1) A549 human pulmonary epithelial cell‐line, (2) human umbilical venous endothelial cells (HUVEC), (3) primary human pulmonary endothelial cells (HPEC) were detected by cell‐based ELISA and indirect immunofluorescence staining. The results revealed that serum levels of IgG anti‐A549 cells antibodies, IgG anti‐HUVEC antibodies, and IgM anti‐HPEC antibodies were significantly higher in SARS patients at phase II/phase III than those in healthy controls. Sera from SARS patients at phase II/phase III could mediate complement dependent cytotoxicity against some A549 cells and HPEC. It is concluded that some autoantibodies against human epithelial cells and endothelial cells would be developed after SARS‐CoV infection and this phenomenon may indicate post‐infectious cellular injury and also induce SARS‐induced immunopathology. J. Med. Virol. 77:1–7, 2005. © 2005 Wiley‐Liss, Inc.
Genetic analysis of human immunodeficiency virus type 1 nef in portugal: Subtyping, identification of mosaic genes, and amino acid sequence variability
Authors: Parreira Ricardo , Pádua Elizabeth , Piedade João , Venenno Teresa , Paixão Maria Teresa , Esteves Aida
Published: July 2005

Extending our previous genetic characterization of human immunodeficiency virus type 1 (HIV‐1) strains circulating in Portugal, we here report the first phylogenetic and putative amino acid sequence variability analyses of nef accessory gene. Viral sequences (n = 53) were amplified by nested PCR from proviral DNA purified from peripheral blood mononuclear cells of HIV‐1 infected individuals (n = 49). Phylogenetic inference analysis demonstrated a distribution of the viral sequences between subtypes A (sub‐subtype A1), B, D, F (sub‐subtype F1), G, H, and J, with subtypes G and B accounting altogether for more than half of the genotypes found. A significant number of the proviral DNA sequences analyzed (18.4%) were shown to correspond to intragenic nef recombinants, with the majority having the typical CRF02_AG nef structure. In addition, three novel intragenic recombinant structures were found (B/G/B, CRF02_AG/H, and D/G). From phylogenetic analysis, it was concluded that part of the non‐recombinant nef genes might have actually been amplified from mosaic viruses: CRF06_cpx, CRF14_BG, and a new envA/nefJ recombinant. While comparing all the putative Nef sequences, significant amino acid sequence variability was observed. However, most of the described nef functional motifs were relatively well conserved in the majority of the sequences analyzed and numerous amino acid changes fell outside these regions. The results presented unambiguously endorse the high level of complexity of HIV‐1 epidemics in Portugal. J. Med. Virol. 77:8–16, 2005. © 2005 Wiley‐Liss, Inc.
Distinct geographic distributions of hepatitis B virus genotypes in patients with acute infection in Japan
Authors: Yotsuyanagi Hiroshi , Okuse Chiaki , Yasuda Kiyomi , Orito Etsuro , Nishiguchi Shuhei , Toyoda Joji , Tomita Eiichi , Hino Keisuke , Okita Kiwamu , Murashima Shiro , Sata Michio , Hoshino Hiromi , Miyakawa Yuzo , Iino Shiro
Published: July 2005

Genotypes of hepatitis B virus (HBV) were determined in 145 patients with acute hepatitis B from various districts in Japan to establish their geographic distribution and evaluating the influence on the clinical illness and outcome. Genotypes were A in 27 (19%) patients, B in 8 (5%), C in 109 (75%) and mixed with B and C in the remaining one (1%). Genotype A was more frequent in metropolitan than the other areas (21/69 (30%) vs. 6/76 (8%), P < 0.001). On phylogenetic analysis, seven of the nine (78%) HBV/A isolates selected at random clustered with those from Europe and the United States, while the remaining two with those of subgroup A' prevalent in Asia and Africa. Maximum ALT levels were lower (2069 ± 1075 vs. 2889 ± 1867 IU/L, P = 0.03) and baseline HBV DNA titers were higher (5.90 ± 1.45 vs. 5.13 ± 1.36 log genome equivalents (LGE)/ml, P = 0.002) in patients infected with genotype A than C. Hepatitis B surface antigen persisted longer in patients infected with genotype A than C (1.95 ± 1.09 vs. 1.28 ± 1.42 months, P = 0.02). HBV infection became chronic in one (4%) patient with genotype A and one (1%) with genotype C infection. Fulminant hepatic failure developed in none of the patients with genotype A, one (13%) with genotype B and five (5%) with genotype C. The point mutation in the precore region (A1896) or the double mutations in the basic core promoter (BCP) region (T1762/A1764) were detected in none of the patients with genotype A, two (25%) with genotype B and 27 (26%) with genotype C. In conclusion, genotype A is frequent in patients with acute hepatitis B in metropolitan areas of Japan, probably reflecting particular transmission routes, and associated with longer and milder clinical course than genotype C. J. Med. Virol. 77:39–46, 2005. © 2005 Wiley‐Liss, Inc.
Full genome analysis of hepatitis B virus genotype E strains from South‐Western Africa and madagascar reveals low genetic variability
Authors: Kramvis Anna , Restorp Karin , Norder Heléne , Botha Johan F. , Magnius Lars O. , Kew Michael C.
Published: July 2005

The eight genotypes of hepatitis B virus (HBV), A–H, have a characteristic geographical distribution. The high prevalence of genotype E within a wide expanse of Africa makes it important to characterize this genotype. Although the preS/S region of a large number of genotype E isolates has been sequenced, to date only eight complete genotype E genomes have been published. The aim of this study, therefore, was to sequence and characterize the complete genome of genotype E isolates obtained from geographically distinct regions in Africa, including Namibia, Angola, and Madagascar. The nine newly sequenced genotype E isolates were closely related with an intra‐group nucleotide divergence of 1.2%, reflecting their close geographical origin. All genotype E strains have the same characteristics, an in‐frame deletion of three nucleotides (one amino acid) in the 5′ pre‐S1, a signature pattern of amino acids in the pre‐S1 region and the serological subtype ayw4. In addition, we have identified another unique attribute of genotype E strains, the introduction of another start codon Met83 in the pre‐S1 region that may result in the translation of an elongated middle hepatitis B surface protein (MHBs). J. Med. Virol. 77:47–52, 2005. © 2005 Wiley‐Liss, Inc.
Oral fluid as a medium for the detection of hepatitis B surface antigen
Authors: Hutse Veronik , Verhaegen Els , De Cock Liesbet , Quoilin Sophie , Vandenberghe Hans , Horsmans Yves , Michielsen Peter , Damme Pierre Van , Vlierberghe Hans Van , Claeys Françoise , Vranckx Robert , Oyen Herman Van
Published: July 2005

Currently viral antigens and antibodies are detected by traditional serological tests. However, the introduction of oral fluid as an alternative medium would allow other alternatives. The collection of oral fluid is, in comparison with venepuncture, less invasive, less painful, less expensive (i.e., no trained personal required), and safe (prevention of needle stick injuries). Also large numbers of samples can be collected easily for epidemiological purposes. Forty‐three HBsAg positive and seventy‐three HBsAg negative paired serum/oral fluid samples were tested. They were collected from patients attending university hospitals. The oral fluid samples were collected using the Oracol collection device and they were subjected to an IgG quantification assay to ensure their quality and quantity. The detection of HBsAg in oral fluid was carried out using a modified ETI‐MAK‐4 ELISA. The validation of this oral fluid test gave a sensitivity and specificity of 90.7% and 100%, respectively. The modified ETI‐MAK‐4 ELISA is a suitable test for oral fluid samples collected by the Oracol collection device for epidemiological purposes. J. Med. Virol. 77:53–56, 2005. © 2005 Wiley‐Liss, Inc.
Pegylated interferon and ribavirin therapy for chronic hepatitis C virus genotype 4 infection
Authors: Legrand‐Abravanel F. , Nicot F. , Boulestin A. , Sandres‐Sauné K. , Vinel J.P. , Alric L. , Izopet Jacques
Published: July 2005

Hepatitis C Virus (HCV) is classified into six genotypes. Genotype 4 is now spreading in Europe, especially among drug users, who are often infected with both HCV and the human immunodeficiency virus (HIV). Previous studies have shown that HCV‐4 responds poorly to interferon. Pegylated interferon (peg‐IFN) associated with ribavirin is now the most effective treatment for eradicating the virus. We have now studied the response of HCV‐4 to peg‐IFN and ribavirin and investigated the influence of HIV infection on anti‐HCV therapy. Twenty‐eight patients infected with HCV‐4 were given peg‐IFN plus ribavirin for 48 weeks. Patients infected with HCV alone tended to have a better initial response (66%) than patients infected with both HCV and HIV (30%, P = 0.06) and eradication was better (50%) than in doubly infected patients (15%, P = 0.06). After controlling for major factors influencing virus response, the virus response 12 weeks after the beginning of treatment in patients infected with HCV‐4 (50%) was similar to that of patients infected with genotype 1 (53%) and lower than that of patients infected with genotypes 2 or 3 (82%, P < 0.05). The response 24 weeks after the end of therapy in patients infected with HCV‐4 (32%) was similar to that of patients infected with HCV‐1 (28%) and lower than that of patients with HCV‐2 or HCV‐3 (62% P < 0.05). These results indicate that HCV‐4 patients should be considered to be difficult‐to‐treat. J. Med. Virol. 77:66–69, 2005. © 2005 Wiley‐Liss, Inc.
Incidence of HCV infection in French hemodialysis units: A prospective study
Authors: Izopet Jacques , Sandres‐Sauné Karine , Kamar Nassim , Salama Gilles , Dubois Martine , Pasquier Christophe , Rostaing Lionel
Published: July 2005

A large prospective study was carried out from 1997 to 2000 in 25 French hemodialysis units including 1,323 patients to determine the incidence of hepatitis C virus (HCV) infection. Monthly testing of alanine aminotransferase (ALT) activity, and assessment of HCV RNA and anti‐HCV antibodies if the ALT activity was elevated, identified 14 new infections in 7 different units, giving an incidence of 0.4% new HCV infections per year. Molecular analyses and epidemiological data indicated that five patients became infected with HCV outside the unit where they were dialyzed, while the nine remaining patients acquired HCV from infected patients on dialysis during the same shift at the same unit. HCV was cleared in six of the seven (85.7%) patients with acute hepatitis C who were given standard doses of alpha‐interferon (α‐IFN). The persistence of nosocomial transmission of HCV in hemodialysis units emphasizes the need to implement infection control practices. Identifying new infections is crucial because α‐IFN treatment results in long term clearance of HCV RNA in a large proportion of patients. J. Med. Virol. 77:70–76, 2005. © 2005 Wiley‐Liss, Inc.
Antigen‐capture enzyme‐linked immunosorbent assay for the diagnosis of crimean‐congo hemorrhagic fever using a novel monoclonal antibody
Authors: Saijo Masayuki , Tang Qing , Shimayi Bawudong , Han Lei , Zhang Yuzhen , Asiguma Muer , Tianshu Dong , Maeda Akihiko , Kurane Ichiro , Morikawa Shigeru
Published: July 2005

An antigen‐capture enzyme‐linked immunosorbent assay (ELISA) was developed for the diagnosis of Crimean‐Congo hemorrhagic fever (CCHF) using a novel monoclonal antibody, 1B7, to the recombinant nucleoprotein (rNP) of CCHF virus (CCHFV) Chinese strain 8402. This ELISA detected at least 2 ng/100 μl of CCHFV rNP of 8402 and of the Nigeria strain Ibr 10200, and also detected authentic nucleoproteins (NP) of Chinese strains. Although the sensitivity of the ELISA was lower than that of nested reverse transcription polymerase chain reaction (RT‐PCR), it was able to detect nucleoproteins in acute sera of CCHF patients. The presence of anti‐CCHFV IgG decreased the sensitivity of the ELISA, possibly due to competition with 1B7, and this would tend to limit application of the ELISA. However, the method may be useful for the diagnosis of CCHF patients in the acute stage of illness, especially in laboratories not equipped with RT‐PCR testing capabilities. J. Med. Virol. 77:83–88, 2005. © 2005 Wiley‐Liss, Inc.
Nitric oxide radical suppresses replication of wild‐type dengue 2 viruses in vitro
Authors: Charnsilpa Weerawan , Takhampunya Ratree , Endy Timothy P. , Mammen Mammen P. , Libraty Daniel H. , Ubol Sukathida
Published: July 2005

Nitric oxide is well accepted as one of the defenses for inhibiting viral dissemination. Macrophages and cells in the macrophage lineage are professional nitric oxide producers which sub‐serve as target for dengue virus. The interaction between nitric oxide and dengue virus in such target cell is unknown. In this report, the impact of nitric oxide on infectious dengue virus serotype 2 production and RNA replication was investigated in vitro. Primary isolates of dengue virus serotype 2 from dengue patients were replicated in mouse neuroblastoma cells in the presence of an exogenous nitric oxide donor, s‐nitroso‐N‐acethylpennicillamine, SNAP, at the concentration of 50 or 75 or 100 μM. Nitric oxide inhibited viral replication in a dose and a multiplicity of infection dependent manner. Nitric oxide from 50 and 75 μM SNAP delayed and suppressed replication of dengue virus isolates while higher concentration of nitric oxide, 100 μM SNAP, completely inhibited production of infectious particles up to 36 hr study. Twenty‐four out of forty tested isolates, 60%, were susceptible to 50 μM SNAP inhibitory effect. The mechanism of inhibition was investigated at the level of RNA synthesis and was found that RNA production was suppressed which correlated to production of the infectious particles. Down‐regulation of the RNA synthesis resulted in reduction of protein synthesis which was detected by lower level of NS1 protein synthesis using immunoblotting. In conclusion, nitric oxide from exogenous nitric oxide donor down regulated replication of dengue virus serotype 2 isolates from dengue patients. The suppression was clearly shown at the level of viral RNA and protein synthesis resulting in reduction of viral progenies production. This phenomenon implies that nitric oxide may serve as a defense which diminishes viral load in patients. J. Med. Virol. 77:89–95, 2005. © 2005 Wiley‐Liss, Inc.