bluuebunny/crossref_metadata_2025 huggingface dataset:
DNA condensation by high‐affinity interaction with avidin
Authors: Morpurgo Margherita , Radu Aurelian , Bayer Edward A. , Wilchek Meir
Published: July 2004

Avidin, the basic biotin‐binding glycoprotein from chicken egg white, is known to interact with DNA, whereas streptavidin, its neutral non‐glycosylated bacterial analog, does not. In the present study we investigated the DNA‐binding properties of avidin. Its affinity for DNA in the presence and absence of biotin was compared with that of other positively charged molecules, namely the protein lysozyme, the cationic polymers polylysine and polyarginine and an avidin derivative with higher isoelectric point (pI ≈ 11) in which most of the lysine residues were converted to homoarginines. Gel‐shift assays, transmission electron microscopy and dynamic light scattering experiments demonstrated an unexpectedly strong interaction between avidin and DNA. The most pronounced gel‐shift retardation occurred with the avidin–biotin complex, followed by avidin alone and then guanidylated avidin. Furthermore, ultrastructural and light‐scattering studies showed that avidin assembles on the DNA molecule in an organized manner. The assembly leads to the formation of nanoparticles that are about 50–100 nm in size (DNA ≈ 5 kb) and have a rod‐like or toroidal shape. In these particles the DNA is highly condensed and one avidin is bound to each 18 ± 4 DNA base pairs. The complexes are very stable even at high dilution ([DNA] =10 pm) and are not disrupted in the presence of buffers or salt (up to 200 mm NaCl). The other positively charged molecules also condense DNA and form particles with a globular shape. However, in this case, these particles disassemble by dilution or in the presence of low salt concentration. The results indicate that the interaction of avidin with DNA may also occur under physiological conditions, further enhanced by the presence of biotin. This DNA‐binding property of avidin may thus shed light on a potentially new physiological role for the protein in its natural environment. Copyright © 2004 John Wiley & Sons, Ltd.
HIV therapy after treatment interruption in patients with multiple failure and more than 200 CD4+ T lymphocyte count
Authors: Jaafar Acil , Massip Patrice , Sandres‐Sauné Karine , Souyris Corinne , Pasquier Christophe , Aquilina Christian , Izopet Jacques
Published: July 2004

The aim of the study was to investigate the safety and efficacy of a salvage therapy initiated after interrupting treatment in patients with virological failure and more than 200 CD4+ T lymphocyte count. In this prospective study, 77 patients who received failing regimens had stopped completely all medication for 3 months before starting an optimised regimen consisting of 3–5 drugs. Patients were tested for HIV resistance before and after treatment interruption. Discontinuation of therapy for 3 months was associated with a median increase in HIV RNA of 1.1 log10, a median decrease in CD4+ T cell count of 136 × 106/L and five clinical events related to HIV, but no AIDS‐defining event. Eighty‐seven percent of patients showed a shift from a drug resistant genotype to a wild‐type genotype based on the major resistance mutations. Forty‐seven percent of patients with a genotype shift reached fewer than 200 HIV RNA copies/ml of plasma 6 and 12 months after treatment resumption whereas none of those without a genotype shift did so (P = 0.03). However, the genotypic shift was not associated with a sustained virological response by multivariate analysis. The use of a new therapeutic class of compound in the salvage regimen was the only predictor of the sustained virological response. Salvage therapy with 3–5 drugs after interrupting treatment for 3 months can be a safe and effective strategy provided the HIV disease is not too advanced. Randomised trials in this population are needed to assess the clinical benefit of this strategy. J. Med. Virol. 74:8–15, 2004. © 2004 Wiley‐Liss, Inc.
Drug resistance mutations and newly recognized treatment‐related substitutions in the HIV‐1 protease gene: Prevalence and associations with drug exposure and real or virtual phenotypic resistance to protease inhibitors in two clinical cohorts of antiretroviral experienced patients
Authors: Torti Carlo , Quiros‐Roldan Eugenia , Monno Laura , Patroni Andrea , Saracino Annalisa , Angarano Gioacchino , Tinelli Carmine , Lo Caputo Sergio , Tirelli Valeria , Mazzotta Francesco , Carosi Giampiero ,   for the GenPheRex and PhenGen Study Groups of the MASTER Cohort
Published: July 2004

This study aimed at identifying HIV‐1 protease amino acid changes associated with protease inhibitor (PI) exposure and susceptibility. New amino acid substitutions were correlated with the number of experienced PIs, reaching statistical significance only for those at positions 3, 44, and 74. The correspondence multivariate model demonstrated that ≥3 experienced PIs and substitutions or mutations at positions 3, 46, 54, 73, 74, and 84 were correlated with PI cross‐resistance, including resistance for lopinavir and amprenavir in this cohort of patients who were naive for these drugs. J. Med. Virol. 74:29–33, 2004. © 2004 Wiley‐Liss, Inc.
HBV and HCV prevalence and viraemia in HIV‐positive and HIV‐negative pregnant women in Abidjan, Côte d'Ivoire: The ANRS 1236 study
Authors: Rouet François , Chaix Marie‐Laure , Inwoley André , Msellati Philippe , Viho Ida , Combe Patrice , Leroy Valériane , Dabis François , Rouzioux Christine ,   for the ANRS 1236 DITRAME‐B&C Study Group
Published: July 2004

A retrospective survey estimating the prevalence of hepatitis viruses B (HBV) and C (HCV) was conducted on samples taken in 1,002 African pregnant women (501 diagnosed as HIV‐1 positive and 501 HIV‐1 negative) participating in a clinical trial program conducted in Abidjan, Côte d'Ivoire (West Africa). Hepatitis B markers studied were HBs antigen (HBsAg), and if positive, HBe antigen/anti‐HBe antibodies and HBV DNA. Two third generation (G3) HCV enzyme immunoassays (EIAs) were used for primary HCV screening. All anti‐HCV antibody‐positive sera were assessed further with supplementary assays (one another G3 EIA, RIBA 3.0, and HCV RNA). HCV genotypes were also determined. HBsAg was found in a similar proportion among HIV‐positive (45/499, 9.0%, 95% confidence interval [95% CI], 6.6–11.9) and HIV‐negative (40/498, 8.0%, 95% CI, 5.8–10.8) women (P = 0.58). The diagnosis of chronic hepatitis B, based on HBV DNA positive results, was more frequent in HIV‐positive women (26.7%), compared to HIV‐negative women (9.4%) (P = 0.06). In the case of hepatitis C infection, after supplementary testing allowing the elimination of frequent false‐positive screening results, a prevalence rate of about 1% was found, both in HIV‐positive (6/501, 1.2%, 95% CI, 0.44–2.59) and HIV‐negative (4/501, 0.8%, 95% CI, 0.22–2.03) women (P = 0.53). Of the 10 samples confirmed positive and assessed for HCV RNA, eight (80%) were viraemic and belonged to HCV genotypes 1 or 2. The relative high frequency of HIV/HBV coinfection in Côte d'Ivoire emphasises the need for monitoring the risk of hepatotoxicity by antiretroviral therapy in such patients. We propose an accurate and cost‐efficient algorithm for HCV diagnosis in Africa. J. Med. Virol. 74:34–40, 2004. © 2004 Wiley‐Liss, Inc.
Genetic variability of hepatitis C virus in chronically infected patients with viral breakthrough during interferon–ribavirin therapy
Authors: Vuillermoz I. , Khattab E. , Sablon E. , Ottevaere I. , Durantel D. , Vieux C. , Trepo C. , Zoulim Fabien
Published: July 2004

Little is known about hepatitis C virus (HCV) breakthrough during antiviral therapy, although it would help in understanding HCV resistance to current antiviral treatments. To analyse the implication of virological factors and the vigour of humoral immune responses in this phenomenon, we studied nine chronic hepatitis C patients with a viral breakthrough during IFN/ribavirin combination therapy, as well as five responders and five non‐responders. The IRES and regions coding for the capsid protein, the PePHD domain of envelope glycoprotein E2 and the NS5A and 5B proteins were amplified by RT‐PCR before treatment, before and during breakthrough, and after treatment. The major variant sequence was obtained by direct sequencing. The heterogeneity of quasispecies was studied by SSCP in all patients and sequencing after cloning in seven genotype 1b‐infected patients. Humoral responses against HCV epitopes were also analysed. The major sequences of IRES, PePHD, and NS5B remained stable during treatment, regardless of the treatment response. However, the capsid protein and the regions flanking PePHD showed sequence variations in breakthrough patients, although no specific mutation was identified. The variable V3 region of NS5A, but not the PKR‐binding domain and the ISDR, seemed to be associated with differences in response to treatment. The analysis of HCV quasispecies revealed no characteristic pattern during treatment in breakthrough patients, whose HCV genome profiles looked most similar to that of non‐responders. The humoral response was similar between groups. In conclusion, viral breakthrough does not seem to be due to selection of resistant strains with signature mutations. J. Med. Virol. 74:41–53, 2004. © 2004 Wiley‐Liss, Inc.
Mutations of the interferon sensitivity‐determining region (ISDR) correlate with the complexity of hypervariable region (HVR)‐1 in the Japanese variant of hepatitis C virus (HCV) type 1b
Authors: Nakano Isao , Fukuda Yoshihide , Katano Yoshiaki , Toyoda Hidenori , Hayashi Kazuhiko , Kumada Takashi , Nakano Satoshi
Published: July 2004

Hepatitis C virus (HCV) genotype 1b comprises mainly two subtypes in Japan, each named for its geographic prevalence (Japan‐specific, J type; worldwide, W type). Because the newly identified subtypes have not been fully characterized, the present study directed this issue from virological viewpoints such as hypervariable region (HVR)‐1 as well as interferon (IFN) sensitivity‐determining region (ISDR). Fifty chronic hepatitis patients with HCV 1b (31 men and 19 women; mean age 50.5 years) were enrolled, and J/W type was determined according to envelope 1 (E1) sequence as described previously (23 J type and 27 W type). Correlations between age, number of HVR‐1 clones, HVR‐1 diversity, and ISDR mutations were analyzed in J and W type patients independently. In addition, the sequences of the three HCV regions obtained for the determination of the above genetic factors were studied phylogenetically. The number of HVR‐1 clones was significantly higher for J type in comparison with W type (P = 0.044). In the J type‐infected patients, the ISDR mutation number was correlated inversely with HVR‐1 clone number (P = 0.0001, r = −0.734) and HVR‐1 diversity (P = 0.0001, r = −0.722). However, this correlation was not observed in the W type patients. W type patients showed a significant correlation between age and HVR‐1 clone number (P = 0.015, r = 0.462). Phylogenetic study revealed that the nonstructural (NS) 5A sequence, which is obtained for ISDR type determination, can distinguish between J and W types. The inverse correlation in J type patients between ISDR mutations and HVR‐1 complexity may explain the usefulness of the ISDR for prediction of IFN response only in Japanese patients. This suggests that the ISDR is not directly related to IFN responsiveness, but the degree of HVR‐1 complexity may be more important. J. Med. Virol. 74:54–61, 2004. © 2004 Wiley‐Liss, Inc.
Hepatitis C virus among self declared non‐injecting sexual partners of injecting drug users
Authors: Roy K.M. , Goldberg D.J. , Hutchinson S. , Cameron S.O. , Wilson K. , MacDonald L.
Published: July 2004

While much is known about hepatitis C virus (HCV) among injecting drug users (IDUs), there is scant information about the risk of HCV infection to non‐injecting sexual partners of injecting drug users; it is possible that such individuals may have a greater risk of acquiring HCV than any other group barring injecting drug users. This study examines the prevalence of HCV among a population of non‐injecting sexual partners of injecting drug users. Unlinked anonymous testing for anti‐HCV of residual sera stored following the named HIV testing of specimens originally from persons who had indicated to their attending clinicians that they were non‐injecting sexual partners of injecting drug users. The prevalence of anti‐HCV among the sexual partners was 4.1% (25/611) overall, 6.4% (13/202) among heterosexual male and 3.0% (12/397) among the heterosexual female partners. None of the homosexual/bisexual partners were HCV antibody positive (0/12). Although we cannot be sure how non‐injecting partners of injecting drug users acquire their HCV infection, having a relationship with someone who injects drugs may place an individual at appreciable risk of being infected; such individuals should consider being tested for HCV. J. Med. Virol. 74:62–66, 2004. © 2004 Wiley‐Liss, Inc.
One‐year survey of astrovirus infection in children with gastroenteritis in a large hospital in Hungary: Occurrence and genetic analysis of astroviruses
Authors: Jakab Ferenc , Meleg Edina , Bányai Krisztián , Melegh Béla , Tímár László , Péterfai János , Szűcs György
Published: July 2004

Human astroviruses (HAstV) are the causative agents of viral gastroenteritis mainly in children worldwide. This study investigated the epidemiology and genotype diversity of HAstVs detected in children admitted to hospital with gastroenteritis in Hungary. Stool samples were collected from children with diarrhea at the Municipal “Szent László” Hospital, Budapest, Hungary, between January 2002 and December 2002. Of 2,758 samples, 607 were negative for both rotaviruses and bacterial pathogens and were tested for astroviruses using a reverse transcriptase‐polymerase chain reaction (RT‐PCR) targeting the open reading frame (ORF2), capsid region. Astrovirus was detected in 10 samples (1.6%) by RT‐PCR. Astrovirus infection was more frequent among children 49 to 60 months of age. Genotyping of positive samples was performed by type‐specific RT‐PCR and confirmed by sequence analysis. Phylogenetic analysis was performed using a 203 nucleotide consensus length of the 3′‐end of the capsid gene. Type‐specific RT‐PCR and sequence analysis detected genotypes 1 (50%), 4 (30%), 3 (10%), and 8 (10%) among the children admitted to hospital. Genotype 1 was the predominant genotype, but genotypes 3, 4, and 8 were also present indicating the importance of emerging genotype 8 infections. Two distinct genotype 4 variants were observed during this study. Sequence analysis confirmed type‐specific RT‐PCR results in the capsid region. This is the first comprehensive report on the occurrence of HAstV infections in Central/Eastern Europe. J. Med. Virol. 74:71–77, 2004. © 2004 Wiley‐Liss, Inc.
Cytomegalovirus antiviral resistance associated with treatment induced UL97 (protein kinase) and UL54 (DNA polymerase) mutations
Authors: Scott G.M. , Isaacs M.A. , Zeng F. , Kesson A.M. , Rawlinson W.D.
Published: July 2004

HCMV‐related illness due to infections with antiviral resistant virus was verified by phenotypic and genotypic assays in 17% (8/47) of high‐risk immunocompromised Australian patients. Selective PCR‐sequencing of UL97 (protein kinase; PK) and UL54 (DNA polymerase; DNApol) regions important for antiviral sensitivity, identified the majority (6/8) of resistant strains through detection of mutations known to confer antiviral resistance. Additionally, eight UL54 (DNApol) mutations (N408K, T691S, A692V, S695T, L737M, A834P, V955I, and A972V) of unknown phenotype were identified in six specimens from patients with clinical evidence of antiviral resistant infections. One isolate was resistant to ganciclovir (GCV) and another resistant to PFA on phenotypic testing where mutations in UL97 (PK) or UL54 (DNApol) were not detected, suggesting a loss of correlation between phenotype and genotype. Selective PCR‐sequencing of UL97 (PK) and UL54 (DNApol) provided rapid and comprehensive results, but missed some resistance detected by phenotypic assays. A combination of phenotypic and genotypic assays is recommended for complete analysis of CMV antiviral resistance, as well as further definition of the clinical relationship between novel UL54 (DNApol) mutations and antiviral resistance. J. Med. Virol. 74:85–93, 2004. © 2004 Wiley‐Liss, Inc.
Detection of high‐risk human papillomavirus E6 and E7 oncogene transcripts in cervical scrapes by nested RT‐polymerase chain reaction
Authors: Sotlar Karl , Stubner Annette , Diemer David , Menton Susanne , Menton Michael , Dietz Klaus , Wallwiener Diethelm , Kandolf Reinhard , Bültmann Burkhard
Published: July 2004

The oncogenic potential of the high‐risk human papillomavirus (HPV) genotypes (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) depends on the expression of the two viral oncogenes E6 and E7. Thus, the detection of HPV E6/E7 oncogene transcripts could serve as a factor in the evaluation of a risk of development of cervical intraepithelial neoplasia (CIN) and its progression to cervical cancer. A nested RT‐PCR assay for the detection of E6/E7 oncogene transcripts of all known high‐risk HPV genotypes was established. In the study described, 779 high‐risk HPV‐DNA‐positive cervical scrapes exhibiting all grades of CIN, including non‐dysplastic cervical mucosa (CIN 0), were examined. Spliced E6/E7 oncogene transcripts of all the high‐risk HPVs were detected in numerous samples, with an overall detection rate of 47%. In 227 cases with agreement between the cytologic and histologic findings, the prevalence increased with lesion severity: CIN 0, 18%; CIN I, 58%; CIN II, 77%; CIN III, 84%. Multiple transcriptionally active high‐risk HPVs were detected in 12% (33/279) of patients with multiple high‐risk HPV infections. This work sets the stage for a prospective follow‐up study currently being undertaken to evaluate the prognostic relevance of the detection of high‐risk HPV E6/E7 oncogene transcripts for the persistence of a high risk HPV infection, and the possible evolution and further development of a CIN. Future applications of the assay described may include the monitoring of women in studies investigating antiviral treatment or vaccination. J. Med. Virol. 74:107–116, 2004. © 2004 Wiley‐Liss, Inc.
Analysis of human papillomavirus type‐16 variants in Italian women with cervical intraepithelial neoplasia and cervical cancer
Authors: Tornesello Maria Lina , Duraturo Maria Luisa , Salatiello Immacolata , Buonaguro Luigi , Losito Simona , Botti Gerardo , Stellato Giovanni , Greggi Stefano , Piccoli Roberto , Pilotti Silvana , Stefanon Bernardina , De Palo Giuseppe , Franceschi Silvia , Buonaguro Franco M.
Published: July 2004

Human papillomavirus type 16 (HPV‐16) classes (E, AA, As, Af1, Af2) and their variants have different geographic distribution and different degrees of association with cervical lesions. This study was designed to examine HPV‐16 variants among Italian women and their prevalence in case patients (affected by invasive cervical carcinoma or cervical intraepithelial neoplasia grade 2–3 and cervical intraepithelial neoplasia grade 1), versus control subjects with normal cervical epithelium (controls). A total of 90 HPV‐16 positive cervical samples from women of Italian Caucasian descent have been tested, including 36 invasive cervical carcinomas, 21 with cervical intraepithelial neoplasias grade 2–3, 17 with cervical intraepithelial neoplasia grade 1 and 16 controls. HPV‐16 was detected with an E6/E7 gene‐specific polymerase chain reaction, and variant HPV‐16 classes and subclasses were identified by direct nucleotide sequencing of the region coding for the E6 and the E7 oncoproteins, the MY09/11‐amplified highly conserved L1 region, and the long control region (LCR). Among the 90 HPV‐16 samples, nine viral variants have been identified belonging to the European (Ep‐T350 and E‐G350) and non‐European (AA and Af‐1) branches. The E‐G350 is the prevalent variant in all analyzed different disease stages being present in 55.5% of ICC, 52.4% of cervical intraepithelial neoplasias 2–3, 47.1% of cervical intraepithelial neoplasia grade 1, and 50.0% of control samples. The non‐European variants AA and Af1, rarely detected in control samples, represent 33.3% of all HPV‐16 infections in invasive cervical carcinoma (with a peak of 19.4% and 13.9%, respectively), showing a statistically significant increase in frequency in more advanced lesions (χ2 trend = 7.2; P < 0.05). The prevalence of HPV‐16 Ep‐T350, however, is higher in controls (43.7%) and in of cervical intraepithelial neoplasia grade 1 (41.2%) than in cervical intraepithelial neoplasia grade 2–3 (28.6%) and in invasive cervical carcinoma (11.1%) cases strongly suggesting lack of progression for pre‐neoplastic lesions associated with such variant. The increased frequency of non‐European variants in invasive lesions suggests that they are more oncogenic than European variants. This could have implications for future diagnostic and therapeutic strategies. J. Med. Virol. 74:117–126, 2004. © 2004 Wiley‐Liss, Inc.
Virus diversity and an outbreak of group C rotavirus among infants and children with diarrhea in Maizuru city, Japan during 2002–2003
Authors: Phan Tung Gia , Nishimura Shuichi , Okame Michio , Nguyen Tuan Anh , Khamrin Pattara , Okitsu Shoko , Maneekarn Niwat , Ushijima Hiroshi
Published: July 2004

A total of 236 fecal specimens collected from infants and children with gastroenteritis in Maizuru city, Japan from July 2002 to June 2003, were tested for the presence of rotaviruses, noroviruses, sapoviruses, astroviruses, and adenoviruses by RT‐PCR, PAGE, RPHA, and latex agglutination methods. Among diarrheal viruses detected, group A rotavirus was the most prevalent (32.2%; 76 of 236) followed by norovirus GII (21.2%; 50 of 236), group C rotavirus (10.2%; 24 of 236), adenovirus (3.8%; 9 of 236), sapovirus (2.5%; 6 of 236), astrovirus (1.3%; 3 of 236), and norovirus GI (0.8%; 2 of 236), respectively. It is noteworthy that group C rotavirus infection was apparently confined only within the period of 5 months (December 2002 through April 2003). This pattern of infection implied that the outbreak of group C rotavirus in these patients, which was the first outbreak of gastroenteritis attributed to group C rotavirus in Maizuru city. Moreover, about half (12 of 24) of group C rotavirus infected cases were confined to infants and young children less than 3 years old. Another interesting feature of the study was the demonstration of the mixed infections with group C rotavirus and group A rotavirus, as well as group C rotavirus and norovirus GII in 20.8% (5 of 24) and 8.3% (2 of 24), respectively. This is the first report of gastroenteritis associated with the mixed infections with group C rotavirus and other viral enteropathogens such as norovirus. The results indicate that group C rotavirus could infect not only older children and adults but also infants and young children under 3 years old. J. Med. Virol. 74:173–179, 2004. © 2004 Wiley‐Liss, Inc.
High‐resolution magic angle spinning and 1H magnetic resonance spectroscopy reveal significantly altered neuronal metabolite profiles in CLN1 but not in CLN3
Authors: Sitter Beathe , Autti Taina , Tyynelä Jaana , Sonnewald Ursula , Bathen Tone F. , Puranen Johanna , Santavuori Pirkko , Haltia Matti J. , Paetau Anders , Polvikoski Tuomo , Gribbestad Ingrid S. , Häkkinen Anna‐Maija
Published: July 2004

The neuronal ceroid lipofuscinoses (NCLs) are among the most severe inherited progressive neurodegenerative disorders of children. The purpose of this study was to compare the in vivo 1.5‐T 1H magnetic resonance (MR) and ex vivo 14.3‐T high‐resolution (HR) magic angle spinning (MAS) 1H MR brain spectra of patients with infantile (CLN1) and juvenile (CLN3) types of NCL, to obtain detailed information about the alterations in the neuronal metabolite profiles in these diseases and to test the suitability of the ex vivo HR MAS 1H MRS technique in analysis of autopsy brain tissue. Ex vivo spectra from CLN1 autopsy brain tissue (n = 9) significantly differed from those of the control (n = 9) and CLN3 (n = 5) groups, although no differences were found between the CLN3 and the control groups. Principal component analysis of ex vivo data showed that decreased levels of N‐acetylaspartate (NAA), γ‐aminobutyric acid (GABA), glutamine, and glutamate as well as increased levels of inositols characterized the CLN1 spectra. Also, the intensity ratio of lipid methylene/methyl protons was decreased in spectra of CLN1 brain tissue compared with CLN3 and control brain tissue. In concordance with the ex vivo data, the in vivo spectra of late‐stage patients with CLN1 (n = 3) revealed a dramatic decrease of NAA and a proportional increase of myo‐inositol and lipids compared with control subjects. Again, the spectra of patients with CLN3 (n = 13) did not differ from those of controls (n = 15). In conclusion, the ex vivo and in vivo spectroscopic findings were in good agreement within all analyzed groups and revealed significant alterations in metabolite profiles in CLN1 brain tissue but not in CLN3 compared with controls. Furthermore, HR MAS 1H MR spectra facilitated refined detection of neuronal metabolites, including GABA, and composition of lipids in the autopsy brain tissue of NCL patients. © 2004 Wiley‐Liss, Inc.
Proteomic analysis of phosphotyrosyl proteins in the rat brain: Effect of butorphanol dependence
Authors: Kim Seong‐Youl , Chudapongse Nuannoi , Lee Sang‐Min , Levin Michael C. , Oh Jae‐Taek , Park Hae‐Joon , Ho Ing K.
Published: July 2004

Butorphanol (17‐cyclobutylmethyl‐3,14‐dihydroxymorphinan) tartrate (Stadol) is a mixed agonist‐antagonist opioid analgesic agent that is about five to seven times as potent as morphine in analgesic effects. The chronic use of butorphanol produces physical dependence in humans and animals. Phosphorylation plays a very important role in developing butorphanol dependence; however, global phosphorylation events induced by chronic butorphanol administration have not been reported. The aim of this study is to determine the alteration of tyrosine phosphorylation of brain frontal cortical proteins in butorphanol‐dependent rats using a proteomic approach. Dependence was produced by continuous intracerebroventricular (i.c.v.) infusion of butorphanol (26 nmol/μl/hr) for 72 hr via osmotic minipump in rats. Similar patterns of protein expression were detected by two‐dimensional electrophoresis (2‐DE) in brain frontal cortex of butorphanol‐dependent and saline‐treated control rats. All 65 phosphotyrosyl (p‐Tyr) protein spots detected in pH 3–10 phosphotyrosine 2‐DE of control rat brains were detected in butorphanol‐dependent rat brains. The densities of most p‐Tyr protein spots were increased in butorphanol‐dependent rat brains compared to saline‐treated control samples. Eighteen additional p‐Tyr protein spots were detected in pH 3–10 2‐DE images of butorphanol‐dependent rat brains. Immobilized pH strips with three different narrow pH ranges were examined to improve the resolution of p‐Tyr proteins in 2‐DE gels. Fifty‐three p‐Tyr protein spots were identified as known proteins involved in cell cytoskeleton, cell metabolism, and cell signaling. This proteomic approach can provide useful information for understanding the complex mechanism of butorphanol dependence in vivo. © 2004 Wiley‐Liss, Inc.
Differential roles of fibroblast growth factor‐2 during development and maintenance of auditory sensory epithelia
Authors: Carnicero Estela , Zelarayan Laura Cecilia , Rüttiger Lukas , Knipper Marlies , Alvarez Yolanda , Alonso Maria Teresa , Schimmang Thomas
Published: July 2004

Fibroblast growth factor‐2 (FGF2) has been postulated to be a key regulator involved in the proliferation, differentiation, and regeneration of sensory hair cells. Here we have addressed the potential functions of FGF2 during the formation and regeneration of the auditory epithelium in chicken and mice. By using viral gene transfer, based on herpes simplex type 1 virus (HSV‐1), we show that ectopically applied FGF2 drastically increases the number of cells expressing early hair cell markers during embryonic development in avians. Intriguingly, FGF2 does not stimulate cell division during this process. These data suggest that FGF2 plays a role during differentiation of sensory hair cells in avians. To address the potential functions of FGF2 during murine inner ear development, we analyzed FGF2 mouse mutants. Mice lacking FGF2 showed normal formation of the inner ear, and no abnormalities were observed at the adult stage. Moreover, FGF2 mouse mutants showed similar hearing thresholds compared with those observed in control mice before and after noise damage. Therefore, endogenous FGF2 appears not to be essential for the development or functional maintenance of the auditory organ in mammals. In light of these results, the differential roles of FGF2 in the vertebrate inner ear are discussed with respect to its previously postulated functions. © 2004 Wiley‐Liss, Inc.
γ‐aminobutyric acidergic interneuron vulnerability to aging in canine prefrontal cortex
Authors: Pugliese M. , Carrasco J.L. , Geloso M.C. , Mascort J. , Michetti F. , Mahy N.
Published: July 2004

The aged dog is considered a promising model for examining molecular and cellular processes involved in a variety of human neurological disorders. By using the canine counterpart of senile dementia of the Alzheimer's type (ccSDAT), we investigated the specific vulnerability of the γ‐aminobutyric acid (GABA) cortical subset of interneurons, characterized by their calcium‐binding protein content, to neuronal death. Dogs representing a large variety of breeds were classified into three groups: young control, aged control, and ccSDAT. In all dogs, the general distribution and cell typology of parvalbumin‐, calretinin‐, and calbindin‐positive neurons were found to be similar to those in the human. As in Alzheimer's disease patients, neurons displaying parvalbumin or calretinin immunoreactivity were resistant and the calbindin‐positive ones depleted. Together with aging, amyloid deposition in its early phase (stage II) participates in this specific neuronal death, but with a lower potency. In conclusion, our data provide evidence that preservation of GABAergic cortical interneurons has to be focused on the early stage of β‐amyloid deposition. We also demonstrate the usefulness of dogs of all breeds for investigating the early phases of human brain aging and Alzheimer's disease. © 2004 Wiley‐Liss, Inc.
Acute and persistent protein synthesis inhibition following cerebral reperfusion
Authors: DeGracia Donald J.
Published: July 2004

Lack of recovery from protein synthesis inhibition (PSI) closely correlates with neuronal death following brain ischemia and reperfusion. It has therefore been suggested that understanding the mechanisms of PSI will shed light on the mechanisms of selective neuronal death following ischemia and reperfusion. It is now known that the PKR‐like ER kinase (PERK)‐mediated phosphorylation of eukaryotic initiation factor 2α (eIF2α) causes translation inhibition at initial reperfusion. Activation of PERK, in turn, indicates endoplasmic reticulum stress and activation of the unfolded protein response. However, phosphorylation of eIF2α is a transient event and can account for PSI only in the initial hours of reperfusion. Although a number of other regulators of protein synthesis, such as eIF4F, 4EBP‐1, eEF‐2, and S6 kinase, have been assessed following cerebral ischemia and reperfusion, the causes of prolonged PSI have yet to be fully elucidated. The purpose of the present article is to bring together the evidence indicating that, at minimum, postischemic PSI should be conceptualized as consisting of two components: an acute, transient component mediated by unfolded protein response‐induced eIF2α phosphorylation and a longer term component that correlates with neuronal death. Ischemic tolerance appears to separate the acute and persistent components of reperfusion‐induced translation inhibition. Specific models of the relationship among acute PSI, persistent PSI, and neuronal death are presented to clarify issues that have emerged from ongoing work in this area. © 2004 Wiley‐Liss, Inc.
Effect of lithium administration on collagen and breaking pressure of the rat thoracic descending aorta
Authors: Tzaphlidou Margaret , Berillis Panagiotis
Published: July 2004

Lithium is widely used in medicine as an antidepressive drug. Despite abundant literature on the subject, questions on the effects of lithium on collagen, which is one of the major components of tissues and organs, are far from being answered. We have examined the effects of lithium chloride (1.5 mEq Li/kg of body weight) on rat thoracic descending aorta: 1) on collagen fibrils by electron microscopy and image analysis and 2) on breaking pressure. Animals were sacrificed 1 day, 2, and 12 months after the end of a 30 consecutive days experimental period. In all cases investigated, structural abnormalities in collagen fibrils, at the ultrastructural level, were found. These abnormalities consist of decreased mean diameter and high variability in width as well as marked disorganization in the packing of the fibrils. The observed alterations seem to be permanent. Rupture of the rat thoracic aorta following pressure is significantly influenced by lithium. J. Trace Elem. Exp. Med. 17:151–160, 2004. © 2004 Wiley‐Liss, Inc.
Zinc deficiency and oxidative stress in brain: Magnetic resonance investigations in weanling rats
Authors: Towner Rheal A. , Appleby Christopher , Levy Mark , Bray Tammy M.
Published: July 2004

In humans, zinc deficiency is characterized by a broad spectrum of neurological clinical syndromes. It is known that vesicular zinc‐enriched areas of the brain, such as the hippocampus, are responsive to zinc deprivation, which may result in learning impairment. Recent findings show that zinc deficiency may cause alterations in neurochemical activity. In this study we used contrast‐enhanced magnetic resonance imaging (MRI) to monitor disruptions to the blood–brain barrier (BBB) and image‐guided MR spectroscopy to follow alterations in brain metabolites as a result of zinc‐deficiency and/or hyperoxia‐induced oxidative stress. Gadolinium‐diethylaminetriaminopentaacetic acid, an extracellular T1 relaxation contrast agent, increases tissue water signal in the brain if the BBB is damaged. A significant increase in postcontrast T1‐weighted MR image intensity was observed in the brain of zinc‐deficient or hyperoxia‐exposed rats, as well as zinc‐deficient rats exposed only to hyperoxia when compared with zinc‐adequate rats. From single‐voxel image‐guided MR spectroscopy results, significant decreases in the ratio of N‐acetyl aspartate, a neuronal‐specific compound, to total choline levels were found when comparing controls (zinc‐adequate or zinc pair‐fed) with zinc‐deficiency or hyperoxia groups alone, and when zinc‐deficiency was combined with hyperoxia. This study demonstrates the sensitivity of MR techniques in the ability to monitor the effect of zinc deficiency combined with oxidative stress on BBB permeability as well as detect alterations in brain metabolites. This will further aid in our understanding of the possible cellular and molecular mechanisms involved in zinc deficiency pathology associated with the brain. J. Trace Elem. Exp. Med. 17:161–174, 2004. © 2004 Wiley‐Liss, Inc.
Well‐posedness and asymptotic analysis for a Penrose–Fife type phase field system
Authors: Rossi Riccarda
Published: July 2004

In this paper, an asymptotic analysis of the (non‐conserved) Penrose–Fife phase field system for two vanishing time relaxation parameters ε and δ is developed, in analogy with the similar analyses for the phase field model proposed by G. Caginalp (Arch. Rational Mech. Anal. 1986; 92:205–245), which were carried out by Rossi and Stoth (Adv. Math. Sci. Appl. 2003; 13:249–271; Quart. Appl. Math. 1995; 53:695–700).

Although formally the singular limits for ε ↓ 0 and for ε and δ ↓ 0 are, respectively, the viscous Cahn–Hilliard equation and the Cahn–Hilliard equation, it turns out that the Penrose–Fife system is indeed a bad approximation for these equations. Therefore, we consider an alternative approximating phase field system, which could be viewed as a generalization of the classical Penrose–Fife phase field system, featuring a double non‐linearity given by two maximal monotone graphs. A well‐posedness result is proved for such a system, and it is shown that the solutions converge to the unique solution of the viscous Cahn–Hilliard equation as ε ↓ 0, and of the Cahn–Hilliard equation as ε ↓ 0 and δ ↓ 0. Copyright © 2004 John Wiley & Sons, Ltd.
Diffusion in poro‐plastic media
Authors: Showalter R. E. , Stefanelli U.
Published: July 2004

A model is developed for the flow of a slightly compressible fluid through a saturated inelastic porous medium. The initial‐boundary‐value problem is a system that consists of the diffusion equation for the fluid coupled to the momentum equation for the porous solid together with a constitutive law which includes a possibly hysteretic relation of elasto‐visco‐plastic type. The variational form of this problem in Hilbert space is a non‐linear evolution equation for which the existence and uniqueness of a global strong solution is proved by means of monotonicity methods. Various degenerate situations are permitted, such as incompressible fluid, negligible porosity, or a quasi‐static momentum equation. The essential sufficient conditions for the well‐posedness of the system consist of an ellipticity condition on the term for diffusion of fluid and either a viscous or a hardening assumption in the constitutive relation for the porous solid. Copyright © 2004 John Wiley & Sons, Ltd.
Time‐dependent algorithms for viscoelastic flow: Finite element/volume schemes
Authors: Webster M. F. , Tamaddon‐Jahromi H. R. , Aboubacar M.
Published: July 2004

Hybrid finite volume/element methods are investigated within the context of transient viscoelastic flows. A finite volume algorithm is proposed for the hyperbolic constitutive equation, of Oldroyd‐form, whereas the continuity/momentum balance is accommodated through a Taylor‐Galerkin finite element method. Various finite volume combinations are considered to derive accurate and stable implementations. Consistency of formulation is key, embracing fluctuation distribution and median‐dual‐cell constructs, within a cell‐vertex discretisation on triangles. In addition, we investigate the effect of treating the time‐term in a finite element fashion, using mass‐matrix iteration instead of the standard finite volume mass‐lumping approach. We devise an accurate transient scheme that captures the analytical solution at short and long time, both in core flow and near shear boundaries. In this respect, some difficulties are highlighted. A new method emerges, with the Low Diffusion B (LDB, with or without mass‐matrix iteration) as the optimal choice. We progress to a complex flow application and demonstrate some provocative features due to the influence of true transient boundary conditions on evolutionary flow‐structure in a 4:1 start‐up rounded‐corner contraction problem. © 2004 Wiley Periodicals, Inc. Numer Methods Partial Differential Eq, 2005
Amifostine for children with medulloblastoma treated with cisplatin‐based chemotherapy
Authors: Fisher Michael J. , Lange Beverly J. , Needle Michael N. , Janss Anna J. , Shu Hui‐Kuo G. , Adamson Peter C. , Phillips Peter C.
Published: July 2004

In adult patients, amifostine appears to ameliorate cisplatin‐related nephrotoxicity and ototoxicity. We assessed the safety and efficacy of amifostine in 11 children with newly diagnosed medulloblastoma/primitive neuroectodermal tumor treated with radiotherapy and vincristine, lomustine, and cisplatin. Amifostine was administered immediately prior to and 4 hr into the cisplatin infusion. Amifostine caused assymptomatic hypotension and hypocalcemia in 18 and 82% of patients, respectively. Despite amifostine use, 78% of patients developed significant ototoxicity. Although relatively well tolerated, amifostine does not appear to have a major impact on ameliorating the risk of developing significant nephro‐ and ototoxicity in children with medulloblastoma. Larger studies will help clarify these findings. Pediatr Blood Cancer 2004;43:780–784. © 2004 Wiley‐Liss, Inc.
A survey of resources and activities in the MISPHO family of institutions in Latin America: A comparison of two eras
Authors: Sala Alessandra , Barr Ronald D. , Masera Giuseppe
Published: July 2004

Background

In developing countries, implementation of effective pediatric cancer programs can reduce the overall rate of childhood mortality. To promote this objective, Monzas International School of Pediatric Hematology‐Oncology (MISPHO)—for 14 countries with limited resources in Latin America—was launched in 1996.

Methods

A survey was conducted to evaluate the possible impact of MISPHO on the following areas: research, educational, and clinical activities, diagnostic services, transfusion medicine, supportive care, ancillary staffing complement, and parents' associations—before and after 1996.

Results

No institution lost resources or activities after 1996 and there were overall gains, in particular in educational activities, diagnostic capabilities, creation of single and multiple institutional treatment protocols.

Conclusions

Although notable improvements have occurred, there are still important needs that have to be satisfied. © 2004 Wiley‐Liss, Inc.
Contact time and interfacial fracture energy of tacky polymers
Authors: Chau Kin W. , Swei Gwo S.
Published: July 2004

The spontaneous adhesion of tacky polymers to rigid materials as a function of contact time at constant temperature and applied pressure is considered. Polymer‐substrate attachment, assumed to initiate as microscopic nuclei that form at random locations on the substrate and expand to cover their surroundings, is studied from a probabilistic perspective. Calculation yields a description of the natural law governing the increase in the interfacial fracture energy of tacky polymers on rigid substrates with contact time. The description assigns to each polymer–substrate pair three parameters that characterize the pair's bonding behavior and serve as a signature for the pair. The proposed description of the underlying natural law is verified with published experimental data. Its main features and implications are examined. The findings pertain to pressure sensitive adhesion and tack, and are used to interpret experimental results. The scheme not only accounts for the reported results of 14 polymer–substrate pairs but also depicts other distinct bonding behaviors yet to be observed. © 2004 Wiley Periodicals, Inc. J Polym Sci Part B: Polym Phys 42: 3013–3025, 2004
Short communication: French version of the Occupational Stress Indicator (OSI): preliminary assessment of reliability and validity
Authors: Steiler Dominique , Cooper Cary L.
Published: July 2004

This article offers a preliminary assessment of the psychometric quality of the French version of the Occupational Stress Indicator (OSI). The OSI is a diagnostic tool that assesses seven different aspects of occupational stress by means of one independent variable (sources of pressure), three moderating factors (Type A behaviour, locus of control and coping strategies), and three dependent variables (job satisfaction, mental health and physical health).

 The purpose of the present study is to test the French translation of the OSI and investigate the reliability of all seven scales, as well as the convergent and discriminant validity of six of them, excluding job satisfaction.

 Cronbach's α revealed that six of the seven OSI scales were statistically very reliable. In accordance with previous studies, the seventh scale, locus of control, proved to be less reliable. Significant and acceptable levels of construct validity were also found for the Type A, locus of control, mental health, physical health scales and for one of the coping strategy subscales: social support.

 These encouraging results should lead to further, broader and more complete assessments of the psychometric quality of the French version of the OSI. Copyright © 2004 John Wiley & Sons, Ltd.
Short communication: Reliability testing and applied use of a measure of neutrophil activation in stress research
Authors: Ellard David R. , Barlow Julie H. , Mian Rubina
Published: July 2004

The purpose of this research was to test the reliability of a measure of neutrophil activation and to apply the measure in stress research. Two studies were carried out with university undergraduates. The first was a test‐retest reliability study of the nitro‐blue tetrazolium (NBT) test, a measure of neutrophil activity. The second was an applied study of its use in stress research. In the first study, finger‐stick blood samples were taken at three points in time (baseline, +1 hour, and at 1 month) from 30 participants. Physiological arousal was monitored (heart rate and blood pressure) before and after samples were taken. Neutrophil activity is determined by their capacity to reduce NBT and is presented as a percentage of NBT positive cells. Highly significant correlations were found between all time points (p = 0.0001) and there were no significant differences between the means. The NBT test was found to be reliable and was used in an examination of the association between neutrophil activity and perceived stress (PSS). Health related issues were also explored. A highly significant positive association was found between NBT and PSS (r = 0.840, p = 0.01). No associations are found between PSS and health. The results confirm and extend findings that stressors, even naturalistic stressors, can activate neutrophils. Copyright © 2004 John Wiley & Sons, Ltd.
ISO/IEC 15504—Evolution to an international standard
Authors: Rout Terence P.
Published: July 2004

This article describes the work currently being undertaken to help ISO/IEC TR 15504 progress to the status of a full International Standard, and outlines the changes in design that are to be incorporated in the revision. It describes the inputs for the design decisions that were taken; identifies the fundamental changes in the architecture of the Standard; and briefly describes the current status of the development of the Standard. Copyright © 2004 John Wiley & Sons, Ltd.
Human peroxiredoxin 5 is a peroxynitrite reductase
Authors: Dubuisson Marlène , Vander Stricht Delphine , Clippe André , Etienne Florence , Nauser Thomas , Kissner Reinhard , Koppenol Willem H , Rees Jean-François , Knoops Bernard
Published: July 2004

Peroxiredoxins are an ubiquitous family of peroxidases widely distributed among prokaryotes and eukaryotes. Peroxiredoxin 5, which is the last discovered mammalian member, was previously shown to reduce peroxides with the use of reducing equivalents derived from thioredoxin. We report here that human peroxiredoxin 5 is also a peroxynitrite reductase. Analysis of peroxiredoxin 5 mutants, in which each of the cysteine residues was mutated, suggests that the nucleophilic attack on the O–O bond of peroxynitrite is performed by the N‐terminal peroxidatic Cys47. Moreover, with the use of pulse radiolysis, we show that human peroxiredoxin 5 reduces peroxynitrite with an unequalled high rate constant of (7 ± 3) × 107 M−1
s−1.