bluuebunny/crossref_metadata_2025 huggingface dataset:
Development and validation of a liquid chromatography and ion spray tandem mass spectrometry method for the quantification of artesunate, artemether and their major metabolites dihydroartemisinin and dihydroartemisinin‐glucuronide in sheep plasma
Authors: Duthaler Urs , Keiser Jennifer , Huwyler Jörg
Published: January 2011

Recently, promising fasciocidal activities of artesunate and artemether were described in rats and sheep. Therefore, a liquid chromatography–tandem mass spectrometry (LC–MS/MS) method was developed to quantify artesunate, artemether and their metabolites dihydroartemisinin and dihydroartemisinin‐glucuronide in sheep plasma. Protein precipitation with methanol was used for sample workup. Reversed‐phase high‐performance liquid chromatography (HPLC) was performed using an Atlantis C18 analytical column with a mobile phase gradient system of ammonium formate and acetonitrile. The analytes were detected by MS/MS using selected reaction monitoring (SRM) with electrospray ionisation in the positive mode (transition m/z 267.4 → 163.0). The analytical range for dihydroartemisinin, dihydroartemisinin‐glucuronide and artesunate was 10–1000 ng/ml and for artemether 90–3000 ng/ml with a lower limit of quantification of 10 and 90 ng/ml, respectively. Inter‐ and intra‐day accuracy and precision deviations were < 10%. Consistent relative recoveries (60–80%) were observed over the investigated calibration range for all analytes. All analytes were stable in the autosampler for at least 30 h (6 °C) and after three freeze and thaw cycles. The validation results demonstrated that the LC–MS/MS method is precise, accurate and selective and can be used for the determination of the artemisinins in sheep plasma. The method was applied successfully to determine the pharmacokinetic parameters of artesunate and its metabolites in plasma of intramuscularly treated sheep. Copyright © 2011 John Wiley & Sons, Ltd.
Investigation of pituitary adenylate cyclase activating polypeptide in human gynecological and other biological fluids by using MALDI TOF mass spectrometry
Authors: Brubel R. , Reglodi D. , Jambor E. , Koppan M. , Varnagy A. , Biro Zs. , Kiss P. , Gaal V. , Matkovits A. , Farkas J. , Lubics A. , Bodis J. , Bay Cs. , Veszpremi B. , Tamas A. , Nemeth J. , Mark Laszlo
Published: January 2011

Pituitary adenylate cyclase activating polypeptide (PACAP) is a multifunctional and pleiotropic neuropeptide. PACAP has diverse effects in the endocrine system, among others, it plays important roles in oogenesis, implantation and development of the nervous system. However, it is not known whether PACAP is present in the fluids of the human reproductive organs. The aim of the present study was to determine, by means of mass spectrometry and radioimmunoassay, whether PACAP is present in human amniotic fluid, ovarian follicular fluid and cervico‐vaginal fluid. Samples were obtained from healthy adult volunteers. Our MALDI TOF and MALDI TOF/TOF spectrometry results show that PACAP38 is present in all of the follicular fluid samples, and PACAP‐like immonoreactivity was also measured by radioimmunoassay. However, we did not find the characteristic peak representing the unmodified 38 amino acid form of the peptide in normal cervico‐vaginal smear and amniotic fluid samples. Furthermore, we analyzed other body fluids for comparison, such as human nasal fluid, saliva and aqueous humor. PACAP was not found in these latter samples. In summary, the present study provides evidence for the presence of PACAP in human follicular fluid, suggesting a role in oocyte function, but determination of the exact physiological significance awaits further investigation. Copyright © 2011 John Wiley & Sons, Ltd.
Sieve‐based device for MALDI sample preparation. III. Its power for quantitative measurements
Authors: Molin Laura , Cristoni Simone , Seraglia Roberta , Traldi Pietro
Published: January 2011

The solid sample inhomogeneity is a weak point of traditional MALDI deposition techniques that reflects negatively on quantitative analysis. The recently developed sieve‐based device (SBD) sample deposition method, based on the electrospraying of matrix/analyte solutions through a grounded sieve, allows the homogeneous deposition of microcrystals with dimensions smaller than that of the laser spot. In each microcrystal the matrix/analyte molar ratio can be considered constant. Then, by irradiating different portions of the microcrystal distribution an identical response is obtained. This result suggests the employment of SBD in the development of quantitative procedures. For this aim, mixtures of different proteins of known molarity were analyzed, showing a good relationship between molarity and intensity ratios. This behaviour was also observed in the case of proteins with quite different ionic yields. The power of the developed method for quantitative evaluation was also tested by the measurement of the abundance of IGPP[Oxi]GPP[Oxi]GLMGPP (m/z1219) present in the collagen‐α‐5(IV) chain precursor, differently expressed in urines from healthy subjects and diabetic‐nephropathic patients, confirming its overexpression in the presence of nephropathy. The data obtained indicate that SBD is a particularly effective method for quantitative analysis also in biological fluids of interest. Copyright © 2011 John Wiley & Sons, Ltd.
Amino acid substitution in the core protein has no impact on relapse in hepatitis C genotype 1 patients treated with peginterferon and ribavirin
Authors: Inoue Yuko , Hiramatsu Naoki , Oze Tsugiko , Yakushijin Takayuki , Mochizuki Kiyoshi , Fukuda Kazuto , Mita Eiji , Haruna Yoshimichi , Inoue Atsuo , Imai Yasuharu , Hosui Atsushi , Miyagi Takuya , Yoshida Yuichi , Tatsumi Tomohide , Kiso Shinichi , Kanto Tatsuya , Kasahara Akinori , Takehara Tetsuo , Hayashi Norio
Published: January 2011

Previous reports demonstrated that amino acid (aa) substitutions in the hepatitis C virus (HCV) core protein are predictors of non‐virological responses to pegylated interferon (Peg‐IFN) and ribavirin combination therapy. The aim of this study was to investigate the impact of core aa substitutions on viral kinetics during the treatment and relapse after the treatment. The 187 patients with HCV genotype 1 enrolled in this study were categorized into four groups according to core aa substitution patterns: double‐wild group (n = 92), Arg70/Leu91; 70‐mutant group (n = 42), Gln70/Leu91; 91‐mutant group (n = 31), Arg70/Met91; and double‐mutant group (n = 22), Gln70/Met91. The relationship between the core aa substitutions and the virological response was examined. Multivariate logistic regression analyses showed that substitution at aa 70 was significantly associated with a poor virological response during the first 12 weeks (decline of <1 log from baseline at week 4, <2 log at week 12), and substitution at aa 91 was significantly associated with detectable HCV RNA at week 24. With respect to relapse, only the ribavirin exposure (odds ratio (OR), 0.77; 95% confidence interval (CI), 0.60–0.98) and HCV RNA disappearance between weeks 13 and 24 (OR, 23.69; 95% CI, 5.44–103.08) were associated independently with relapse, with no correlation being found with the core aa substitutions and relapse. In conclusion, the results showed that core aa substitutions can be strong predictive factors at pretreatment of the non‐response, but not for relapse, for virological responders with HCV RNA disappearance during treatment. J. Med. Virol. 83:419–427, 2011. © 2011 Wiley‐Liss, Inc.
Influence of HCV genotype 1 subtypes on the virus response to PEG interferon alpha‐2a plus ribavirin therapy
Authors: Nicot F. , Alric L. , Barange K. , Métivier S. , Dramard J.M. , Combis J.M. , Castan B. , Meurisse J.J. , Payen J.L. , Garipuy D. , Desmorat H. , Peron J.M. , Thebault S. , Morin T. , Renou C. , Barel P. , Guerin B. , Imbert Y. , Sire S. , Sauné K. , Chatelut E. , Izopet J.
Published: January 2011

New factors that influence the viral response in HCV non‐genotype 2/3 patients must be identified in order to optimize anti‐HCV treatment. This multicenter prospective study evaluates the influence of HCV variability and pharmacological parameters on the virological response of these patients to pegylated interferon α2a (peg‐IFN‐α2a: 180 µg/week) and ribavirin (RBV; 800–1,200 mg/day) for 48 weeks. HCV subtypes were identified by sequencing the NS5B region. Serum RBV and peg‐IFN‐α2a concentrations were measured at weeks 4 and 12. The 115 patients (67 men; median age = 49, range 31–76) included 64 who had never been treated and 27 co‐infected with HIV. The mean baseline HCV RNA was 6.30 ± 0.06 log IU/ml and the HCV genotypes were: G1 (n = 93) with 1a (n = 37) and 1b (n = 50), G4 (n = 20) and G5 (n = 2). Most patients (79/108; 73%) had an early virological response. Independent predictors of an early virological response were interferon naive patients (OR = 2.98, 95% CI: 1.15–7.72) and RBV of >2,200 ng/ml at week 12 (OR = 3.41, 95% CI: 1.31–8.90). Forty of 104 patients (38%) had a sustained virological response. The only independent predictors of a sustained virological response were subtype 1b (OR = 6.82, 95% CI: 1.7–26.8), and HCV RNA <15 IU/ml at week 12 (OR = 25, 95% CI: 6.4–97.6). Thus a serum RBV concentration of >2,200 ng/ml was associated with an early virological response and patients infected with HCV subtype 1b had a better chance of a sustained virological response than did those infected with subtype 1a. J. Med. Virol. 83:437–444, 2011. © 2011 Wiley‐Liss, Inc.
Sequencing and phylogenetic analysis of the coding region of six common rotavirus strains: Evidence for intragenogroup reassortment among co‐circulating G1P[8] and G2P[4] strains from the United States
Authors: Bányai K. , Mijatovic‐Rustempasic S. , Hull J.J. , Esona M.D. , Freeman M.M. , Frace A.M. , Bowen M.D. , Gentsch J.R.
Published: January 2011

The segmented genome of rotaviruses provides an opportunity for rotavirus strains to generate a large genetic diversity through reassortment; however, this mechanism is considered to play little role in the generation of mosaic gene constellations between Wa‐like and DS‐1‐like strains in genes other than the neutralization antigens. A pilot study was undertaken to analyze these two epidemiologically important strains at the genomic level in order to (i) identify intergenogroup reassortment and (ii) to make available additional reference genome sequences of G1P[8] and G2P[4] for future genomics analyses. The full or nearly complete coding region of all 11 genes for 3 G1P[8] (LB2719, LB2758, and LB2771) and 3 G2P[4] (LB2744, LB2764, and LB2772) strains isolated from children hospitalized with severe diarrhea in Long Beach, California, where these strains were circulating at comparable rates during 2005–2006 are described in this study. Based on the full‐genome classification system, all G1P[8] strains had a conserved genomic constellation: G1‐P[8]‐I1‐R1‐C1‐M1‐A1‐N1‐T1‐E1‐E1‐H1 and were mostly identical to the few Wa‐like strains whose genome sequences have already been determined. Similarly, the genome sequences of the 3 G2P[4] strains were highly conserved: G2‐P[4]‐I2‐R2‐C2‐M2‐A2‐N2‐T2‐E2‐E2‐H2 and displayed an overall lesser genetic divergence with reference DS‐1‐like strains. While intergenogroup reassortment was not seen between the G1P[8] and G2P[4] strains studied here, evidence for intragenogroup reassortment events was identified. Similar studies in the post‐rotavirus genomic era will help uncover whether intergenogroup reassortment affecting the backbone genes could play a significant role in any potential vaccine breakthrough events by evading immunity of vaccinated children. J. Med. Virol. 83:532–539, 2011. © 2011 Wiley‐Liss, Inc.
Prevalence of human papillomavirus among women with normal cervical cytology in Kuwait
Authors: Al‐Awadhi Rana , Chehadeh Wassim , Kapila Kusum
Published: January 2011

This study was undertaken to determine the prevalence and type specific distribution of human papillomavirus (HPV) in women with normal cervical cytology in Kuwait. The study is the first of its type in Kuwait and one of few in the Middle East. The age specific distribution of HPV types was determined in 3,011 ThinPrep samples taken from women seeking routine gynaecological care. ThinPrep samples were screened for HPV DNA by real‐time PCR. The type specific distribution of the viruses was determined by PCR‐based sequencing. The results showed that HPV DNA was detected in 71 women (2.4%), and 21 different HPV genotypes were detected, comprising eight high‐risk (HR) (16, 31, 33, 53, 56, 58, 66, and 73), seven low‐risk (LR) (6, 11, 54, 61, 70, 81, and 90), four intermediate‐risk (IR) (67, 82, 83, and 84) and HPV 102 and HPV 106. LR HPV types were found in 71.8% of infected samples, HR types in 32.3%, and IR types in 7%. With regard to age, 40.8% of all HPVs were found in women 30–39 years of age, 29.6% in women 40–49 years of age, 19.7% in women over 50 years and 9.9% in women less than 34 years old. The study shows that the prevalence of HPV infection in Kuwait is among the lowest in the world and suggests that HPV vaccine could prevent the development of HPV associated cervical cancer in 1.39% of young females living in Kuwait. However, more extensive population‐based studies should be undertaken before implementing HPV vaccination. J. Med. Virol. 83:453–460, 2011. © 2011 Wiley‐Liss, Inc.
Phylogenetic investigation of nosocomial transmission of hepatitis C virus in an oncology ward
Authors: Dencs Ágnes , Hettmann Andrea , Martyin Tibor , Jekkel Csilla , Bányai Tivadar , Takács Mária
Published: January 2011

Nosocomial hepatitis C virus (HCV) infections have been reported from different health‐care settings worldwide. Twenty patients, treated at the same oncology department, with no previous record of hepatitis C infection, tested positive for anti‐HCV antibodies between November 2007 and June 2008. Twelve of the newly infected patients were found to be HCV RNA positive. The common origin of the infections was assumed. To investigate the relatedness of the detected viral strains phylogenetic analyses were performed using sequences from the NS5B and E1/E2 genome regions. A patient carrying HCV for years was also involved in the study. She was treated at the same oncology department and was considered a possible infectious source. The previous HCV carrier harbored subtype 1b, while all other patients were infected with subtype 1a. Sequences from the 12 newly infected patients formed two groups. The viral sequences within the groups were very closely related. A greater evolutionary distance was observed between the two groups; however, their relatedness could be demonstrated by sequences from both regions with high statistical support. The results indicated that nosocomial transmission occurred. The phylogenetic analyses suggested that the viruses originated from a common source, possibly a patient carrying highly divergent variants. This presumed infectious source could not be identified in the course of this study. The genotype distribution of Hungarian control sequences included in the analysis confirmed this conclusion, since HCV genotype 1a was found to be relatively uncommon. J. Med. Virol. 83:428–436, 2011. © 2011 Wiley‐Liss, Inc.
Hantavirus infection during a stay in a mountain hut in Northern Slovakia
Authors: Zelena Hana , Zvolankova Vlasta , Zuchnicka Jana , Liszkova Katerina , Papa Anna
Published: January 2011

Hantaviruses in Europe cause human hemorrhagic fever with renal syndrome (HFRS) with various degree of severity. The most severe form is caused by the Dobrava/Belgrade virus (DOBV), associated with the rodent Apodemus flavicollis. During the last decade cases of infection caused by DOBV have been reported in Central Europe. The present study is a report on two Czech patients with severe HFRS who were infected during their stay in a mountain hut in Northern Slovakia. The two patients, combined with a third case observed in the same year in a nearby village in the Czech Republic, suggest that this region in Central Europe has to be considered as endemic for HFRS. J. Med. Virol. 83:496–500, 2011. © 2011 Wiley‐Liss, Inc.
Prevalence of norovirus GII‐4 antibodies in Finnish children
Authors: Nurminen K. , Blazevic V. , Huhti L. , Räsänen S. , Koho T. , Hytönen V.P. , Vesikari T.
Published: January 2011

Noroviruses (NoVs) are the second most common cause of viral gastroenteritis after rotavirus in children. NoV genotype GII‐4 has emerged as the major type not only in outbreaks of NoV gastroenteritis but also endemic gastroenteritis among infants and young children worldwide. Using baculovirus‐insect cell system virus‐like particles (VLPs) of NoV genotype GII‐4 and an uncommon genotype GII‐12 were produced. These VLPs were used in enzyme‐linked immunosorbent assays (ELISA) for detection of NoV‐specific immunoglobulin G (IgG) and IgA antibodies in 492 serum specimens from Finnish children 0–14 years of age collected between 2006 and 2008. NoV IgG antibody prevalence was 47.3% in the age group 7–23 months and increased up to 91.2% after the age of 5 years. Avidity of NoV IgG antibodies was low in the primary infections while high avidity antibodies were detected in the recurrent infections of the older children. In GII‐4 infections, the homologous antibody response to GII‐4 VLPs was stronger than to GII‐12 VLPs but cross‐reactivity between GII‐4 and GII‐12 was observed. Binding of GII‐4 VLPs to a putative carbohydrate antigen receptor H‐type 3 could be blocked by sera from children not infected with NoV during a waterborne outbreak of acute gastroenteritis. Therefore, protection against NoV infection correlated with strong blocking activity. J. Med. Virol. 83:525–531, 2011. © 2011 Wiley‐Liss, Inc.
Correlation of polymerase replication fidelity with genetic evolution of influenza A/Fujian/411/02(H3N2) viruses
Authors: Wong Karen Ka Yin , Bull Rowena A. , Rockman Steve , Scott Gillian , Stelzer‐Braid Sacha , Rawlinson William
Published: January 2011

Influenza virus evolves continuously through mutations presumed to result from evolutionary pressure driving viral replication. This study examined the relationship between the genetic evolution and replication fidelity of influenza viruses. Analysis of influenza sequences from National Centre for Biotechnology Information (NCBI) database revealed a gradual decrease in the rate of genetic evolution of A/Fujian/411/02(H3N2)‐like variants after the emergence and predominance of the A/H3N2 Fujian strain in 2002. This decrease may be related to an increase in replication fidelity, which was investigated by assessing mutation frequencies of reassortant viruses carrying the PB1 segment of Fujian variants isolated between 2003 and 2005 in a sequencing‐based plaque assay. The data revealed a threefold decrease in substitution per site of the reassortant viruses carrying the Fujian PB1 segments isolated in 2004–2005 compared with those circulating in 2003. The decrease in mutation frequency paralleled a decrease in genetic evolution of the Fujian variants from the NCBI database. This correlation implicates changes in the polymerase replication fidelity as contributing to altered genetic evolution of influenza viruses. J. Med. Virol. 83:510–516, 2011. © 2011 Wiley‐Liss, Inc.
Analytical and clinical performances of a restriction fragment mass polymorphism assay for detection and genotyping of a wide spectrum of human papillomaviruses
Authors: Lee Hyo‐Pyo , Kim Soo‐Ok , Hwang Tae Sook , Bae Jae‐Man , Kim Soo Nyung , Kim Jae Won , Hwang Sun Young , Lee Han Sung , Shin Soo‐Kyung , Cho Woojae , Hong Sun Pyo
Published: January 2011

Matrix‐assisted laser desorption ionization time‐of‐flight (MALDI‐TOF) mass spectrometry‐based restriction fragment mass polymorphism (RFMP) assay was adapted to human papillomavirus (HPV) genotyping. The analytical sensitivity and the clinical utility were evaluated by testing defined HPV genome equivalents and a total of 426 specimens composed of normal cytology, atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion, high grade squamous intraepithelial lesion and invasive squamous cell carcinoma. The RFMP assay was able to detect 38.4–114.6 genomic equivalents of a wide variety of HPV types. The RFMP assay detected 34 different HPV genotypes in cervical samples of which 8% were found to be multiple‐type infections. The high‐risk HPV positivity rate according to the histological diagnosis was 7.9% (8/101), 31.7% (38/120), 50% (55/110), 86% (37/43), 96.2% (50/52) in normal, atypical squamous cells of undetermined significance, low grade squamous intraepithelial lesion, high grade squamous intraepithelial lesion and squamous cell carcinoma subgroups, respectively. Diagnostic sensitivities/specificities for the cervical lesions of squamous cell carcinoma and high grade squamous intraepithelial lesion or worse histology were found to be 96.2%/92.1% and 91.6%/92.1%, respectively. The sensitivity, accuracy, wide range of genotype identification and high‐throughput capacity with cost‐effectiveness of the test consumables make the RFMP assay suitable for mass screening and monitoring of HPV‐associated cervical cancer. J. Med. Virol. 83:471–482, 2011. © 2011 Wiley‐Liss, Inc.
Prevaccination genomic diversity of human papillomavirus genotype 11: A study on 63 clinical isolates and 10 full‐length genome sequences
Authors: Maver Polona J. , Kocjan Boštjan J. , Seme Katja , Potočnik Marko , Gale Nina , Poljak Mario
Published: January 2011

Prevaccination genomic diversity of human papillomavirus genotype 11 (HPV 11) was established by sequencing 40% of the genome of 63 clinical isolates obtained from an ethnogeographically closed Caucasian cohort, and full‐length genome sequencing of the ten most divergent isolates. In the study, which included the largest number of isolates to date, by analyzing pooled L1, LCR, E6, E5a, and E5b sequences (3,217 bp) of an individual isolate, a total of 23 genomic variants were identified, of which three (5 isolates) and twenty (58 isolates) corresponded to prototypic and non‐prototypic variant groups, respectively. Several novel, potentially important mutations are described. Full‐length genome sequences of ten isolates revealed more than 99% similarity to the HPV 11 prototype isolate. The minimum genomic distance between the full‐length sequences of genomic variants and the prototype was 3 point mutations and 2 inserts and the maximum distance 31 point mutations, one insertion and one deletion. Within the ethnogeographically closed cohort investigated in this study, HPV 11 was shown to be less polymorphic in comparison to the majority of HPV genotypes studied to date. J. Med. Virol. 83:461–470, 2011. © 2011 Wiley‐Liss, Inc.
Algorithm based on CMV kinetics DNA viral load for preemptive therapy initiation after hematopoietic cell transplantation
Authors: Halfon Philippe , Berger Pierre , Khiri Hacene , Martineau Agnès , Pénaranda Guillaume , Merlin Michèle , Faucher Catherine
Published: January 2011

Preemptive therapy in hematopoietic cell transplantation is initiated by a diagnostic technique at first detection of cytomegalovirus (CMV). The aim of this study was to use the viral dynamics of CMV DNA viral to start preemptive therapy, and to prospectively compare the CMV viral load kinetics to pp65 antigenemia. Two hundred sixty‐three blood samples were collected prospectively from 93 patients. All clinical decisions regarding use of preemptive therapy were based on CMV antigenemia. Based on the positivity of the antigen assay and clinical CMV outcome in allotransplant patients, an optimal threshold of 3.05 log 10 (1,130 copies/ml) was found to discriminate patients who required preemptive therapy and those who did not (sensitivity, 71%; specificity, 65%). A DNAemia level increase of 2.24 log 10 (174 copies/ml) per day was the optimal threshold to discriminate between patients who required preemptive therapy and those who did not (sensitivity, 93%; specificity, 43%). Sensitivity of PCR assay was 92.4% compared with 39% for the antigen assay (P < 0.001). A standardized real‐time PCR assay is more appropriate than the antigen assay for detecting CMV. It allowed earlier diagnosis of active CMV infection and monitoring of the response to anti‐CMV treatment. J. Med. Virol. 83:490–495, 2011. © 2011 Wiley‐Liss, Inc.
Characterization of HIV‐1 from patients with virological failure to a boosted protease inhibitor regimen
Authors: Lillemark Marie Rathcke , Gerstoft Jan , Obel Niels , Kronborg Gitte , Pedersen Court , Jørgensen Louise Bruun , Madsen Tina Vasehus , Katzenstein Terese Lea
Published: January 2011

The use of highly active antiretroviral treatment (HAART) regimens with unboosted protease inhibitors (PIs) has resulted in a high level of virological failure primarily due to the development of resistant virus. Current boosted PI regimens combine successfully low‐dose ritonavir (r) with a second PI. The aim of the study was to estimate the proportion of patients, in a population based setting, who develop virological failure on a PI/r regimen. Through The Danish HIV Cohort Study 1,007 patients who received PI/r based treatment between 1995 and 2008 were identified. Twenty‐three (2.3%) experienced virological failure, of whom 19 (83%) started PI/r treatment before 2001. Patients from Copenhagen (n = 19) were selected to study the development of protease (PR) and gag cleavage site (CS) mutations during PI/r treatment and PI plasma levels at the time of virological failure. Three patients (16%) developed major PI resistance mutations. Mutations in the p7/p1 and p1/p6 gag CS only developed in patients with major or minor mutations in PR. Drug concentrations were low or undetectable in 10 out of the 19 patients. In total PR resistance mutations and low drug levels could account for 12 (63%) of the failure cases. In conclusion, virological failure to PI/r is a low and decreasing problem primarily caused by low plasma drug levels and to a lesser extent major PR mutations. Gag CS mutations did not contribute significantly to resistance development and virological failure. J. Med. Virol. 83:377–383, 2011. © 2011 Wiley‐Liss, Inc.
Coxsackievirus B3‐associated aseptic meningitis: An emerging infection in Hong Kong
Authors: Wong Ann H. , Lau C.S. , Cheng Peter K.C. , Ng Anita Y.Y. , Lim Wilina W.L.
Published: January 2011

Enterovirus (EV) infection is a common disease of childhood and associated not uncommonly with aseptic meningitis. In the summer of 2008, laboratory surveillance has detected increased number of coxsackievirus B3 (CVB3) associated aseptic meningitis in Hong Kong, constituting 11.6% of those infected. This study analyzed the epidemiology, circulating pattern, and clinical presentations of CVB3 in Hong Kong over the last 10 years with reference to the circulation of EV in the locality. Enteroviruses (EV) were isolated from respiratory, cerebrospinal fluid (CSF), stool, and vesicular samples using human rhabdomyosarcoma, human laryngeal carcinoma (HEp2‐C), human lung fibroblast (MRC‐5), and African green monkey kidney (Vero) cell lines. Virus isolates were identified and characterized by indirect immunofluorescence (IF) using monoclonal antibodies (mAB), neutralization test as well as partial VP1 sequencing. Different from previous years, IF test result showed that majority of the isolates from 2008 were untypeable by the mAB suggesting antigenic change. Sequence analysis revealed that these isolates were clustered with recent isolate from Fuyang, China. Review of data from 1999 to 2008 showed increased activity of CVB3 in the years 2005 and 2008, and isolates in these 2 years displayed an amino acid change from threonine to alanine at codon 277 of the VP1 gene, which may be associated with central nervous system (CNS) disease. J. Med. Virol. 83:483–489, 2011. © 2011 Wiley‐Liss, Inc.
Epidemiological and phylogenic study of human metapneumovirus infections during three consecutive outbreaks in Normandy, France
Authors: Legrand Loïc , Vabret Astrid , Dina Julia , Petitjean‐Lecherbonnier Joëlle , Stéphanie Gouarin , Cuvillon Delphine , Tripey Valérie , Brouard Jacques , Freymuth François
Published: January 2011

Human metapneumovirus (hMPV) is responsible for respiratory tract disease, particularly in the young and elderly population. An epidemiological and phylogenic study was performed on children admitted to hospital with an acute lower respiratory tract infection (LRI). Data were obtained and analyzed over three consecutive winters, from 2002–2003 to 2004–2005. Each year during the winter period, from November to March, 2,415 nasal swabs were tested by a direct immunofluorescence assay (DFA) for influenza viruses A and B, respiratory syncytial virus, parainfluenza viruses, and adenoviruses. Rhinoviruses, enteroviruses, and coronaviruses OC43 and 229E were detected by RT‐PCR. A RT‐PCR designed for the M gene was performed on negative samples for hMPV detection and phylogenic analyses. For the three consecutive winters, hMPV represented 10%, 22.6%, and 8.8% of virus‐negative samples, respectively. In most cases, clinical symptoms indicated a LRI with a final diagnosis of bronchiolitis. During the winter of 2003–2004, all viral clusters (A1, A2, B1, and B2) that circulated in France shifted progressively from the A group to the B group. This study determined the prevalence of hMPV in Normandy, its clinical impact and permitted the analysis of the molecular evolution during the successive outbreaks. J. Med. Virol. 83:517–524, 2011. © 2011 Wiley‐Liss, Inc.
Effects of Piriformospora indica and Sebacina vermifera on growth and yield of essential oil in fennel (Foeniculum vulgare) under greenhouse conditions
Authors: Dolatabadi Hossein Kari , Goltapeh Ebrahim Mohammadi , Jaimand Kamkar , Rohani Neda , Varma Ajit
Published: January 2011

Fennel (Foeniculum vulgare) is a very important plant in the family of Apiaceae. Effects of inoculation of two endophytic fungi (Piriformospora indica and Sebacina vermifera) in growth, yield and composition of the essential oil of fennel (F. vulgare) were evaluated in pot cultures. Dry fruits were ground with an electric grinder and oil was extracted by hydrodistillation, and their composition was determined by GC/MS. In pot experiment, the maximum dry weight of the green tissue and root and plant height were obtained with P. indica, and maximum number of umbels per plant and dry weight of 1000 fruits were produced with S. vermifera. The P. indica and S. vermifera inoculation significantly increased oil yield as compared to non‐inoculated control plants. GC and GC/MS studies revealed that the level of anethole was increased with P. indica and S. vermifera. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Isolation of nine Phytophthora capsici pectin methylesterase genes which are differentially expressed in various plant species
Authors: Li Peiqian , Feng Baozhen , Wang Hemei , Tooley Paul W. , Zhang Xiuguo
Published: January 2011

Phytophthora capsici causes damage on many plants species, and secretes various pectin methylesterases during all stages of infection. We identified nine Pme genes (Pcpme 1–9) from a genomic library of highly virulent P. capsici strain SD33 and further analyzed the expression pattern of nine genes on three hosts: pepper, tomato, and cucumber using qRT‐PCR during all stages of infection. All nine genes were found to be differentially expressed in three host species in the course of P. capsici interaction. The expression levels of the respective genes increased from 1 to 7 dpi in pepper, while most genes presented a decreasing trend of expression from 1 to 5 dpi in tomato fruits. However, in both fruits peaks were reached at 7 dpi. In cucumber fruits, each gene showed minor expression levels from 1 to 3 dpi, exhibited definite peaks at 5 dpi, and then decreased from 5 to 7 dpi. Thus, evidence from our studies of Pcpme gene expression in different plants at a rang of time points suggests that the late stages of infection may represent the most critical time for P. capsici to successfully express or/and secret PMEs. (© 2011 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)
Highly Efficient Drug Delivery Nanosystem via L‐Phenylalanine Triggering Based on Supramolecular Polymer Micelles
Authors: Dong Haiqing , Li Yongyong , Wen Huiyun , Xu Meng , Liu Lijian , Li Zhuoquan , Guo Fangfang , Shi Donglu
Published: January 2011

An intelligent drug delivery nanosystem has been developed based on biodegradable supramolecular polymer micelles (SMPMs). The drug release can be triggered from SMPMs responsively by a bioactive agent, L‐phenylalanine in a controlled fashion. The SMPMs are constructed from ethylcellulose‐graft‐poly(ε‐caprolactone) (EC‐g‐PCL) and α‐cyclodextrin (α‐CD) derivate via host–guest and hydrophobic interactions. It has been found that these SMPMs have disassembled rapidly in response to an additional L‐phenylalanine, due to great affinity discrepancy to α‐CD between L‐phenylalanine and PCL. Experiments have been carried out on trigger‐controlled in vitro drug release of the SMPMs loaded with a model porphyrin based photosensitizer THPP. The result shows that the SMPMs released over 85% THPP in 6 h, which is two orders magnitudes faster than that of control. Also investigated is the photodynamic therapy (PDT) of THPP‐loaded SMPMs with and without L‐phenylalanine on MCF‐7 carcinoma cell line. An effective trigger‐concentration dependent lethal effect has been found showing promise in clinical photodynamic therapy.

magnified image
Towards Chiral Microporous Soluble Polymers—Binaphthalene‐Based Polyimides
Authors: Ritter Nicola , Senkovska Irena , Kaskel Stefan , Weber Jens
Published: January 2011

Recently, intrinsically microporous polymers have received increased interest in research. Contrary to currently commercially used microporous materials, e.g., zeolithes or polymer networks, they can be dissolved, processed and further functionalised. Microporous poly(imide)s are especially interesting, as they are chemically and thermally resistant to decomposition up to high temperatures. In this paper, we report for the first time on a chiral poly(imide) which is intrinsically microporous and soluble in common solvents. After analysing the structure of the polymer, its pore system is characterised via its ability to adsorb nitrogen, argon, hydrogen and carbon dioxide. Furthermore, the chiral polymer is compared with its racemic counterpart, illustrating the special role of an organised superstructure.

magnified image
Targeting PKC‐β II by Peptides and Peptidomimetics Derived from RACK 1: An In Silico Approach
Authors: Jain Kapil , Sobhia M. Elizabeth
Published: January 2011

Diabetes causes contractile myocardial dysfunction through accelerated atherosclerosis and hypertension, termed Diabetic Cardiomyopathy. Experimental results reveals liaison between cardiovascular disease and diabetic complications. Protein kinase C (PKC), a heterogeneous family of phospholipid dependent kinases was found to be specifically involved in diabetic complications, of which PKC‐β II isoform played a significant role in induction of this fatal disease. Members of PKC family share high degree of similarity in both structure and functions, which has given rise to specificity related issues. In the present study, we have designed peptides and peptidomimetics based on RACK 1 (Receptor for Activated C Kinases) region, as this protein increases the substrate phosphorylation and stabilizes the activated form of PKC‐β II. RACK 1 being specific for PKC‐β II could resolve the specificity issue and also peptides and peptidomimetics, being conformationally restrained structures offers potential advantages of being used as drug molecules over organic molecules. This study has provided useful insights that may contribute to the development of molecules which may be useful in the treatment of diabetic complications.
Individual and combined effects of subclinical doses of deoxynivalenol and fumonisins in piglets
Authors: Grenier Bertrand , Loureiro‐Bracarense Ana‐Paula , Lucioli Joelma , Pacheco Graziela Drociunas , Cossalter Anne‐Marie , Moll Wulf‐Dieter , Schatzmayr Gerd , Oswald Isabelle P.
Published: January 2011

Scope:Deoxynivalenol (DON) and fumonisins (FB) are the most frequently encountered mycotoxins produced byFusariumspecies and most commonly co‐occur in animal diets. These mycotoxins were studied for their toxicity in piglets on several parameters including plasma biochemistry, organ histopathology and immune response.

Methods and results:Twenty‐four 5‐wk‐old animals were randomly assigned to four different groups, receiving separate diets for 5 wk, a control diet, a diet contaminated with either DON (3 mg/kg) or FB (6 mg/kg) or both toxins. At days 4 and 16 of the trial, the animals were subcutaneously immunized with ovalbumin to assess their specific immune response. The different diets did not affect animal performance and had minimal effect on hematological and biochemical blood parameters. By contrast, DON and FB induced histopathological lesions in the liver, the lungs and the kidneys of exposed animals. The liver was significantly more affected when the two mycotoxins were present simultaneously. The contaminated diets also altered the specific immune response upon vaccination as measured by reduced anti‐ovalbumin IgG level in the plasma and reduced lymphocyte proliferation upon antigenic stimulation. Because cytokines play a key role in immunity, the expression levels of IL‐8, IL‐1β, IL‐6 and macrophage inflammatory protein‐1β were measured by RT‐PCR at the end of the experiment. The expression of these four cytokines was significantly decreased in the spleen of piglets exposed to multi‐contaminated diet.

Conclusion:Taken together, our data indicate that ingestion of multi‐contaminated diet induces greater histopathological lesions and higher immune suppression than ingestion of mono‐contaminated diets.
RAFT Inverse Miniemulsion Polymerization of Acrylic Acid and Sodium Acrylate
Authors: Ouyang Liu , Wang Lianshi , Schork F. Joseph
Published: January 2011

The synthesis of well‐controlled poly(sodium acrylate) by RAFT inverse miniemulsion polymerization is presented. The effects of RAFT agent concentration, pH of the dispersed phase, types of initiator, and initiator concentration on the polymerization were studied. The partial loss of control at pH above 7 was likely due to the hydrolysis of the RAFT agent. Particle nucleation was investigated by GPC. It was found that the molecular weight distribution was unimodal when using the lipophilic initiator AIBN. Chain extension was carried out, and the morphology of the final latex (after chain extension) was analyzed by SEM. The reason for the wide particle size distribution observed is discussed.

magnified image
Algebraic multilevel iteration method for lowest order Raviart–Thomas space and applications
Authors: Kraus J. K. , Tomar S. K.
Published: January 2011

An optimal order algebraic multilevel iterative method for solving system of linear algebraic equations arising from the finite element discretization of certain boundary value problems, that have their weak formulation in the space H(div), is presented. The algorithm is developed for the discrete problem obtained by using the lowest‐order Raviart–Thomas space. The method is theoretically analyzed and supporting numerical examples are presented. Furthermore, as a particular application, the algorithm is used for the solution of the discrete minimization problem which arises in the functional‐type a posteriori error estimates for the discontinuous Galerkin approximation of elliptic problems. Copyright © 2011 John Wiley & Sons, Ltd.
Non‐reflecting boundary conditions for acoustic propagation in ducts with acoustic treatment and mean flow
Authors: Redon E. , Dhia A.‐S. Bonnet‐Ben , Mercier J.‐F. , Sari S. Poernomo
Published: January 2011

We consider a time‐harmonic acoustic scattering problem in a 2D infinite waveguide with walls covered with an absorbing material, in the presence of a mean flow assumed uniform far from the source. To make this problem suitable for a finite element analysis, the infinite domain is truncated. This paper concerns the derivation of a non‐reflecting boundary condition on the artificial boundary by means of a Dirichlet‐to‐Neumann (DtN) map based on a modal decomposition. Compared with the hard‐walled guide case, several difficulties are raised by the presence of both the liner and the mean flow. In particular, acoustic modes are no longer orthogonal and behave asymptotically like the modes of a soft‐walled guide. However, an accurate approximation of the DtN map can be derived using some bi‐orthogonality relations, valid asymptotically for high‐order modes. Numerical validations show the efficiency of the method. The influence of the liner with or without mean flow is illustrated. Copyright © 2011 John Wiley & Sons, Ltd.
Development of a quadratic finite element formulation based on the XFEM and NURBS
Authors: Haasemann G. , Kästner M. , Prüger S. , Ulbricht V.
Published: January 2011

The FE‐simulation of inhomogeneous structures, such as composite materials, biological tissues or foams, requires the generation of respective finite element meshes. With increasing complexity of the inner architecture of such structures, this becomes a time‐consuming and laborious task. Additionally, the risk of forming bad‐shaped elements that may lead to ill‐conditioned numerical problems grows significantly. A solution to this problem provides the extended finite element method (XFEM). Thereby, the interface between different materials is represented by a local enrichment of the displacement approximation. As a consequence of this, the element boundary need not be aligned to the interface.

In order to improve the accuracy of the interface approximation, the development of a plane element based on the XFEM and quadratic shape functions will be presented. This element allows for the description of curved material interfaces. The computation of the element stiffness matrix requires a numerical integration process that accounts for discontinuous fields. Regarding a linear element formulation, this can be achieved by an adapted triangulation of the element domain. However, in the case of a curved interface this solution is not applicable. Hence, non‐uniform rational B‐Spline (NURBS) surfaces are used to evaluate the integrals numerically.

Finally, the results of different examples will show the general properties such as the accuracy of the numerical integration procedure and the convergence behavior of this element formulation. Copyright © 2011 John Wiley & Sons, Ltd.
Enriched contact finite elements for stable peeling computations
Authors: Sauer Roger A.
Published: January 2011

During peeling of a soft elastic strip from a substrate, strong adhesional forces act locally inside the peeling zone. It is shown here that when a standard contact finite element (FE) formulation is used to compute the peeling process, a large mesh refinement is required since the numerical solution procedure becomes unstable otherwise. To improve this situation, several different efficient enrichment strategies are presented that provide stable solution algorithms for comparably coarse meshes. The enrichment is based on the introduction of additional unknowns inside the contact elements discretizing the slave surface. These are chosen in order to improve the approximation of the peeling forces, while keeping the overall number of degrees of freedom low. If needed, these additional unknowns can be condensed out locally. The enrichment formulation is developed for both 2D and 3D nonlinear FE formulations. The new enrichment technique is applied to the peeling computation of a gecko spatula. The proposed enriched contact element formulations are also investigated in sliding computations. Copyright © 2011 John Wiley & Sons, Ltd.
Taphonomic modifications produced by modern brown bears (Ursus arctos)
Authors: Saladié P. , Huguet R. , Díez C. , Rodríguez‐Hidalgo A. , Carbonell E.
Published: January 2011

Actualism has been a fundamental tool in taphonomy. The knowledge of accumulation patterning of modern faunal allows us to interpret the activity of different actors in the archaeological record and to reconstruct the behaviour of preterit animals and humans in which we are interested. However, until now, there are few works that include bone modifications made by bears amongst those made by carnivores.

Most data about bone modifications made by bears have been obtained from the archaeo‐palaeontological record. In most of these assemblages, the presence of bears is related to their period of hibernation. Therefore, in these contexts, the changes documented on recovered bear bones are associated only with cannibalism.

In this paper, we present an actualistic study about modifications on bones made by modern brown bears. These animals can cause damage similar to those produced by other large carnivores. Generally, bear activity leaves slight damage, mainly on large‐sized animal bones. However, on bones of small‐sized animals and those of greater fragility, the bears can produce abundant damage. Though not usual, bears can break long bones and consume the complete epiphysis. This study suggests that bears have the potential to be agents of bone modification in fossil assemblages. Consequently, they should be considered as a possible agent of modification of faunal remains in the fossil record. Copyright © 2011 John Wiley & Sons, Ltd.
Following the white rabbit: A case of a small game procurement site in the upper palaeolithic (Sala de las Chimeneas, Maltravieso Cave, Spain)
Authors: Rodríguez‐Hidalgo A. J. , Saladié P. , Canals A.
Published: January 2011

The role of small game in Upper Palaeolithic economies has been examined by several authors over the past few decades. Among the small fauna, rabbits seem to have been a widely exploited resource in certain geographic areas, making them one of the main sources for subsistence in hunter–gatherer groups of southwestern Europe. Examining aspects such as anatomic profiles, demographics and taphonomic modifications (including bone breakage) allows us to approach determining the origins of the accumulation of assemblages and establishing the role of hominids and other processes in their formation.

The faunal remains of level A of the Sala de las Chimeneas (Maltravieso Cave, Extremadura, Spain) show a clear predominance of rabbits. Results point to a complex taphonomic history. The assemblage of rabbit remains recovered at the site is of anthropic, exogenous and intrusive origin.

The results of anthropogenic modifications of the bones reveal a different pattern than what is usually found in assemblages of the same age. The differences that set this assemblage apart include the absence of burned bones and the scarcity of anthropic breakage of humeri, femora and tibiae. These features, in conjunction with the characteristics of the archaeological context, have allowed us to rule out the possibility that this location was used as a base camp or bivouac. Along with these data, the presence of Pleistocene age warrens in the deposit supports the hypothesis that the space was used by hominids as a resource procurement site. Copyright © 2011 John Wiley & Sons, Ltd.