bluuebunny/crossref_metadata_2025 huggingface dataset:
Molecular epidemiology of hepatitis C genotype 6a from patients with chronic hepatitis C from Hong Kong
Authors: Li Cheryl S.Y. , Chan Paul K.S. , Tang Julian W.
Published: February 2009

There have been relatively few studies on HCV genotype 6 compared to hepatitis C virus (HCV) genotype 1. In Hong Kong, a city of about 7 million people, most patients infected with HCV are infected with either genotype 1b or 6a. It is known that HCV 6 is of intermediate responsiveness (i.e., between that of HCV genotype 1 and HCV genotypes 2/3) to standard combination interferon/ribavirin therapy. This study examines the molecular epidemiology of chronic HCV 6a infection in Hong Kong during 1999–2005, as well as characterizing some pre‐ and post‐treatment changes in the NS5B gene in a few patients that underwent combination treatment. Partial non‐structural protein sequences (NS5B, the viral RNA‐dependent RNA polymerase gene, positions 397–1,082) were cloned and sequenced in samples from 51 patients that were obtained and archived during 1999–2005. There were three patients with paired pre‐ and post‐treatment samples available for further analysis. Within this NS5B sequence, one significant amino acid mutation was found in each of these patients: Ser272Pro (end‐of‐treatment response but relapsed), Met312Thr (failed treatment after 3 months due to anemia), and Arg221Lys (end‐of‐treatment‐response but relapsed). This analysis of the pre‐ and post‐treatment NS5B sequences, suggests that whilst post‐treatment mutations were identifiable in individual patients, there was no overall characteristic mutation pattern associated with treatment that was common to all treated patients, identified in this small study. Larger studies are required to confirm these findings. J. Med. Virol. 81:628–633, 2009 © 2009 Wiley‐Liss, Inc.
Humoral immunity in natural infection by tick‐borne encephalitis virus
Authors: Venturi Giulietta , Martelli Paola , Mazzolini Elena , Fiorentini Cristiano , Benedetti Eleonora , Todone Daniele , Villalta Danilo , Fortuna Claudia , Marchi Antonella , Minelli Giada , Ciufolini Maria Grazia
Published: February 2009

Tick‐borne encephalitis (TBE) virus is one of the most important flaviviruses associated with neurological disease in Europe. Cross‐reactive antibodies elicited by different flaviviruses can make difficult the interpretation of ELISA and hemagglutination‐inhibition (HI) tests for the diagnosis of TBE. Neutralization tests, which are more specific, are not in common use because they are difficult to perform and standardize. A plaque reduction neutralization test (PRNT), optimized previously in vaccinated children, was evaluated in sera from acute cases of TBE, collected for diagnostic purposes, and from healthy human population and wild ruminants, collected for serosurvey purposes. The PRNT results were compared with the results of ELISA and HI tests. In acute TBE disease, most sera were positive for IgM antibodies by ELISA and with high HI antibody titers; neutralizing antibodies were detected in 71.4% of patients, at a very low titer (1:10 NT50) in almost all cases. Seroprevalences of 8% and 6.5% for anti‐TBE ELISA antibodies were found in healthy subjects and wild ruminants, respectively. Among anti‐TBE positive healthy subjects, a very low 1:10 NT50 titer was detected in 17.4% of cases, while NT80 titers ranging from 1:10 to 1:80 were detected in 65.2% of cases. Among wild ruminants, 90.9% of ELISA and HI positive samples showed a positive, ≥1:10 NT80 titer. In conclusion, neutralization assays can be useful for the diagnosis and serosurveys of TBE. J. Med. Virol. 81:665–671, 2009 © 2009 Wiley‐Liss, Inc.
Genetic variability and phylogeny of the high‐risk HPV‐31, ‐33, ‐35, ‐52, and ‐58 in central Brazil
Authors: Raiol Tainá , Wyant Patrícia Soares , de Amorim Regina Maria Santos , Cerqueira Daniela Marreco , Milanezi Natália von Gal , Brígido Marcelo de Macedo , Sichero Laura , Martins Cláudia Renata Fernandes
Published: February 2009

More than 100 HPV types have been described, 13 of which are classified as high‐risk due to their association with the development of cervical cancer. The intratype genomic diversity of HPV‐16 and ‐18 has been studied extensively, while little data have been generated for other less common high‐risk types. The present study explores the nucleotide variability and phylogeny of the high‐risk HPV‐31, ‐33, ‐35, ‐52, and ‐58, in samples from Central Brazil. For this purpose, the LCR and the E6 and L1 genes were sequenced. Several variants of these HPV types were detected, some of which have been detected in other parts of the world. Furthermore, new variants of all types examined were characterized in a total of 13 new variants. Based on the E6 and L1 sequences, variants were described comprising conservative and non‐conservative amino acid changes. For phylogenetic tree construction, samples characterized in this study were compared to others described and submitted to GenBank previously. The phylogenetic analysis of HPV‐31, ‐33, ‐35, and ‐58 isolates did not reveal ethnic or geographical clustering as observed previously for HPV‐16 and ‐18. HPV‐35 analysis showed a dichotomic branching characteristic of viral subtypes. Interestingly, four clusters relative to the analysis of HPV‐52 isolates were identified, two of which could be classified as Asian and European branches. The genomic characterization of HPV variants is crucial for understanding the intrinsic geographical relatedness and biological differences of these viruses and contributes further to studies on their infectivity and pathogenicity. J. Med. Virol. 81:685–692, 2009 © 2009 Wiley‐Liss, Inc.
High prevalence of non‐B HIV‐1 subtypes in seamen and their sexual partners in Croatia
Authors: Ramirez‐Piedad Martha Kathleen , Lepej Snježana Židovec , Yerly Sabine , Begovac Josip
Published: February 2009

Travel influences the spread of non‐B subtypes of HIV in Europe. The aim of this study was to assess whether the spread of non‐B subtypes occurred in Croatia, a country with a low level HIV infection. Phylogenetic analysis of the reverse transcriptase and protease genes was undertaken in 145 Croatian patients with the following risks for HIV‐infection: men who have sex with men (n = 64), heterosexual contact (n = 54), intravenous drug use (n = 21), bisexual contact (n = 4) and intravenous drug use plus heterosexual exposure (n = 2). The sample included 25 seamen and 13 steady female sexual partners of seamen. Overall, 38 (26%) of HIV‐patients were infected with non‐B subtypes. Non‐B subtype infections were found only in patients with a possible heterosexual exposure. Non‐B infections were most frequent in seamen and their regular female partners (in 27 of 38 seamen and their partners, 71%). The relative distribution of non‐B subtypes was as follows: A, 8%; C, 26%; F, 3%; CRF01_AE, 8%; CRF02_AG, 34%; CRF05_DF, 5%; CRF06_cpx, 3%; CRF10_CD 8%; and subtype U, 5%. There were altogether 19 heterosexual couples, phylogenetic analysis confirmed that the paired samples were related in all cases. Non‐B subtypes were found only in individuals with a heterosexual risk, mainly in seamen and their partners. J. Med. Virol. 81:573–577, 2009 © 2009 Wiley‐Liss, Inc.
Seroepidemiology of hepatitis A among Greek children indicates that the virus is still prevalent: Implications for universal vaccination
Authors: Kyrka A. , Tragiannidis A. , Cassimos D. , Pantelaki K. , Tzoufi M. , Mavrokosta M. , Pedeli X. , Athanassiadou F. , Hatzimichael A. , Konstantopoulos A. , Kafetzis D. , Papaevangelou V.
Published: February 2009

A national cross‐sectional seroprevalence survey was conducted in order to evaluate the current seroepidemiology of hepatitis A among 1,383 children, aged 0–14 years, residing in Greece. Stratification of the study population was conducted according to age and area of residence. Sera from study participants were tested for the presence of anti‐HAV IgG antibodies. Immigrant children, as well as children residing in rural areas, had lower immunization rates. Among unvaccinated children, the seroprevalence rate of anti‐HAV was 17.1%. Nationality was shown to have a marginally significant effect since non‐immunized immigrant children had a higher seroprevalence rate (22.4% vs. 15.9%, OR = 1.52, P = 0.064). Significant differences between geographic areas for both vaccination coverage and natural immunity were observed. The study findings indicate that hepatitis A is prevalent in Greece and therefore universal infant hepatitis A immunization should be implemented. J. Med. Virol. 81:582–587, 2009 © 2009 Wiley‐Liss, Inc.
Detection of HPV types and neutralizing antibodies in Gansu province, China
Authors: Wu Xueling , Zhang Chuntao , Feng Shuxian , Liu Chunyu , Li Yanqin , Yang Yongxiu , Gao Jun , Li Hongfang , Meng Shufang , Li Liping , Zhang Yunzhong , Hu Xuemei , Wu Xiaolu , Lin Lin , Li Xun , Wang Youchun
Published: February 2009

A total of 82 samples from patients with cervical cancer (Group 1) and 50 samples from patients with other genital diseases (Group 2) were collected in Gansu, China. All 132 samples were tested for HPV DNA with a typing kit that can detect 21 types of HPV, and also tested for neutralizing antibodies against HPV‐16, ‐18, ‐58, ‐45, ‐6, and ‐11 using pseudovirus‐based neutralization assays. The results revealed that 28% (23/82) of sera in Group 1 were positive for type‐specific neutralizing antibodies with a titer range of 160–640, of which 23.2% (19/82), 2.4% (2/82), 2.4% (2/82), 1.2% (1/82), and 1.2% (1/82) were against HPV‐16, ‐58, ‐6, ‐18, and ‐45, respectively. Only one serum (2%) in Group 2 was positive for neutralizing antibodies, which were against HPV‐6 with a titer of 2,560. Overall, 85.4% (70/82) of samples in Group 1 were HPV DNA‐positive, compared with 28% (14/50) of samples in Group 2. The seven most common types detected in Group 1 were HPV‐16 (80%), HPV‐52 (7.1%), HPV‐66 and HPV‐11 (5.7% each), and HPV‐58, HPV‐18, and HPV‐33 (4.3% each), while the four most common types in Group 2 were HPV‐16 (12%), HPV‐52 and HPV‐11 (6% each), and HPV‐68 (4%). The concordance between HPV DNA and corresponding neutralizing antibodies was 32.9% (27/82) with a significant difference (P < 0.005). More specifically, the concordance was 42.7% (35/82) for HPV‐16 in Group 1. The full‐length sequences of six HPV types (HPV‐16, ‐58, ‐33, ‐59, ‐11, and ‐68) were determined and showed 99% identities with their reported genomes. J. Med. Virol. 81:693–702, 2009 © 2009 Wiley‐Liss, Inc.
Randomized trial of high‐dose interferon‐α‐2b combined with ribavirin in patients with chronic hepatitis C: Correlation between amino acid substitutions in the core/NS5A region and virological response to interferon therapy
Authors: Mori Nami , Imamura Michio , Kawakami Yoshiiku , Saneto Hiromi , Kawaoka Tomokazu , Takaki Shintaro , Aikata Hiroshi , Takahashi Shoichi , Chayama Kazuaki
Published: February 2009

The aim of this study was to compare the efficacy of high‐dose interferon (IFN)‐α‐2b with standard dose of IFN‐α‐2b in combination with ribavirin (RBV) for patients with chronic hepatitis C virus (HCV) infection, and to investigate the predictive factors associated with virological response. Two hundred Japanese patients with high HCV viral load (>100 KIU/ml) were randomized to 6 or 10 mega units (MU) of 24‐week IFN‐α‐2b with RBV. Predictive factors were investigated; including pretreatment amino acid (aa) sequences of the core region and the IFN‐sensitive determining region (ISDR). The sustained virological response rate was not different in the two groups (24% vs. 30%) but the incidence of depression was significantly higher in the 10 MU group than 6 MU group (7% vs. 0%, P = 0.02). Younger age (<60) and HCV genotype (2a/b) were significant predictors of sustained virological response. In patients infected with genotype 1b, substitutions of core aa 70 and/or 91 were predictive for non‐virological response (P < 0.001), and substitutions in the ISDR was observed frequently in virological responders. Early viral kinetics study showed that serum HCV core antigen decreased more slowly in both patients with aa 70 and/or 91 substitutions in the core and with absence of substitutions in the ISDR. In conclusion, the use of a higher dose of IFN‐α‐2b in combination with RBV did not improve virological response but resulted in higher incidence of depression. Amino acid substitutions in the core and ISDR are predictive of virological response to the therapy in patients with genotype 1b and high viral load. J. Med. Virol. 81:640–649, 2009 © 2009 Wiley‐Liss, Inc.
Phylogenetic analysis of novel G12 rotaviruses in the United States: A molecular search for the origin of a new strain
Authors: Freeman M.M. , Kerin T. , Hull J. , Teel E. , Esona M. , Parashar U. , Glass R.I. , Gentsch J.R.
Published: February 2009

Rotavirus serotype G12 was initially identified in the Philippines in 1987 and was not described again until it reemerged more than 13 years later. G12 strains were first detected in the United States in 2002 and have recently assumed a worldwide distribution. The high similarity between the sequence of the major outer capsid VP7 gene of human G12 strains and the single porcine G12 isolate raised the prospect that human strains may have arisen through reassortment with porcine strains or, alternatively, that the porcine strain originally came from humans. We sequenced portions of the remaining 10 segments of two human G12 strains (G12P[8] and G12P[6]) and a currently circulating common strain (G1P[8]) identified during the 2005–2006 surveillance season and compared the sequences with those of strains available through GenBank. By comparison, the three strains were all Wa‐like and not porcine‐like. A newly outlined classification system proposed genotypes for each gene segment based on nucleotide similarity. Using this approach, gene segments VP1–3, VP6 and NSP1–5 grouped within the same genotype, indicating that the three strains analyzed were closely related. These results suggest that the novel G12P[8] strain could have been formed by the solitary introduction of a VP7 gene into a globally common rotavirus strain, G1P[8]. Classifying rotavirus strains based only on VP7 (G) and VP4 (P) genotype potentially underestimates diversity and sequence analysis of the other segments is required to assess the complete genetic relationships between strains. J. Med. Virol. 81:736–746, 2009 © 2009 Wiley‐Liss, Inc.
Hepatitis B e antigen‐negative mutations in the precore and core promoter regions in Korean patients
Authors: Choi Jong Won , Ahn Sang Hoon , Park Jun Yong , Chang Hye Young , Kim Ja Kyung , Baatarkhuu Oidov , Kim Do Young , Han Kwang‐Hyub , Chon Chae Yoon
Published: February 2009

Most patients with hepatitis B e antigen (HBeAg)‐negative chronic hepatitis B have variants of the hepatitis B virus (HBV) that include mutations in the precore or core promoter regions of the HBV genome. The aim of this study was to investigate the patterns of precore and core promoter mutations and their relationship to HBeAg expression in Korean patients. Four hundred seventy‐five Korean patients with chronic HBV infection between February 1995 and December 2003 were enrolled in this study. There were 236 HBeAg‐positive and 239 HBeAg‐negative patients. Blood samples were tested for HBsAg, anti‐HBs, HBeAg, hepatitis B e antibody (anti‐HBe), liver function tests, and serum HBV DNA. Mutations in the precore and core promoter regions were determined by direct sequencing. In the core promoter region, the C1740, C1753, T1762/A1764, and T1766 mutations were associated with HBeAg escape (all; P < 0.05). In the precore region, a higher frequency of the C1802, A1828, T1846, A1850, C1858, T1862, and A1896 mutations was found in HBeAg‐negative patients (all; P < 0.05). In particular, the A1896 mutation was associated with high serum levels of ALT and HBV DNA in HBeAg‐negative patients (P = 0.014 and 0.026, respectively). Mutations around the Kozak sequence (nucleotides 1809–1812) were found in 6.7% of patients and were not associated with undetectable HBeAg (P = 0.13). In Korean patients, various mutations in the precore and core promoter regions were associated with HBeAg escape and amelioration of hepatic inflammation in HBeAg‐ negative patients. Only the A1896 mutation contributed to HBeAg‐negative chronic hepatitis B. J. Med. Virol. 81:594–601, 2009 © 2009 Wiley‐Liss, Inc.
Combined therapy of interferon plus ribavirin promotes multiple adaptive solutions in hepatitis C virus
Authors: Cuevas José M. , Torres‐Puente Manuela , Jiménez‐Hernández Nuria , Bracho María A. , García‐Robles Inmaculada , Carnicer Fernando , Olmo Juan del , Ortega Enrique , González‐Candelas Fernando , Moya Andrés
Published: February 2009

Hepatitis C virus (HCV) presents several regions involved potentially in evading antiviral treatment and host immune system. Two regions, known as PKR‐BD and V3 domains, have been proposed to be involved in resistance to interferon. Additionally, hypervariable regions in the envelope E2 glycoprotein are also good candidates to participate in evasion from the immune system. In this study, we have used a cohort of 22 non‐responder patients to combined therapy (interferon alpha‐2a plus ribavirin) for which samples obtained just before initiation of therapy and after 6 or/and 12 months of treatment were available. A range of 25–100 clones per patient, genome region and time sample were obtained. The predominant amino acid sequences for each time sample and patient were determined. Next, the sequences of the PKR‐BD and V3 domains and the hypervariable regions from different time samples were compared for each patient. The highest levels of variability were detected at the three hypervariable regions of the E2 protein and, to a lower extent, at the V3 domain of the NS5A protein. However, no clear patterns of adaptation to the host immune system or to antiviral treatment were detected. In summary, although high levels of variability are correlated to viral adaptive response, antiviral treatment does not seem to promote convergent adaptive changes. Consequently, other regions must be involved in evasion strategies likely based on a combination of multiple mechanisms, in which pools of changes along the HCV genome could confer viruses the ability to overcome strong selective pressures. J. Med. Virol. 81:650–656, 2009 © 2009 Wiley‐Liss, Inc.
Primary vasculitis of the central nervous system in patients infected with HIV‐1 in the HAART era
Authors: Melica Giovanna , Brugieres Pierre , Lascaux Anne‐Sophie , Levy Yves , Lelièvre Jean‐Daniel
Published: February 2009

Angiitis of the central nervous system (CNS) in patients infected with HIV‐1‐is often associated with concomitant infection or lymphoproliferative disease of the CNS. Four HAART naïve patients infected with HIV‐1 with severe stroke are described. Evidence of vasculitis was found by magnetic resonance angiography. Extensive investigations excluded concomitant opportunistic, lymphoproliferative or autoimmune disorders leading to the diagnosis of primary angiitis of the CNS. Despite initiation of HAART and prolonged suppression of viral replication, these patients remained severely immunosuppressed. The addition of corticosteroids led to a significant improvement of clinical symptoms. Primary angiitis of the CNS should be considered in patients with HIV and stroke. The prognosis of these patients remain poor despite HAART. These observations suggest that the vascular inflammatory process persists despite the control of viral load under HAART in patients with persistent immunosuppression. J. Med. Virol. 81:578–581, 2009 © 2009 Wiley‐Liss, Inc.
Interleukin‐4 blocks thapsigargin‐induced cell death in rat rod photoreceptors: Involvement of cAMP/PKA pathway
Authors: Adão‐Novaes Juliana , Guterres Céulem de Cássia Belem , da Silva Ana Gabriela Ledo Santos , Campello‐Costa Paula , Linden Rafael , Sholl‐Franco Alfred
Published: February 2009

Although the photoreceptors cell death is the main cause of some retinopathies diseases, the mechanisms involved in this process are poorly understood. The neuroprotective effects of interleukin‐4 (IL‐4) have been shown in several tissues, including retina. We demonstrate that treatment of rat retinal explants with IL‐4 completely inhibited the thapsigargin‐induced rod photoreceptor cell death after 24 hr in culture. We also showed that IL‐4 receptor α subunit (IL‐4Rα) is abundantly present in retina. Colocalization of IL‐4Rα and rhodopsin indicate a direct effect of this cytokine in rod photoreceptor cells. Moreover, IL‐4 increased the intracellular levels of cAMP in 7.4‐fold, indicating that the neuroprotective effect of this cytokine was completely blocked by RpcAMP, an inhibitor of protein kinase (PKA). Our data demonstrate, for the first time, the neuroprotective effect of IL‐4 through cAMP/PKA pathway in thapsigargin‐induced photoreceptor cell death. © 2009 Wiley‐Liss, Inc.
Induction of GAP‐43 modulates neuroplasticity in PBSC (CD34+) implanted‐Parkinson's model
Authors: Shyu Woei‐Cherng , Li Kuo‐Wei , Peng Hsiao‐Fen , Lin Shinn‐Zong , Liu Ren‐Shyan , Wang Hsiao‐Jung , Su Ching‐Yuan , Lee Yih‐Jing , Li Hung
Published: February 2009

As a result of the progressive decrease in efficacy of drugs used to treat Parkinson's disease (PD) and the rapid development of motor complications, effective alternative treatments for PD are required. In a 6‐hydroxydopamine (6‐OHDA)‐induced Parkinson's rat model, intracerebral peripheral blood stem cell (CD34+) (PBSC) transplantation significantly protected dopaminergic neurons from 6‐OHDA‐induced neurotoxicity, enhanced neural repair of tyrosine hydroxylase neurons through up‐regulation of Bcl‐2, facilitated stem cell plasticity, and attenuated activation of microglia, in comparison with vehicle‐control rats. The 6‐OHDA‐lesioned hemi‐Parkinsonian rats receiving intrastriatal transplantation of PBSCs also showed: 1) enhanced glucose metabolism in the lesioned striatum and thalamus, demonstrated by [18F]fluoro‐2‐deoxyglucose positron emission tomography (FDG‐PET), 2) improved neurochemical activity as shown by proton magnetic resonance spectroscopy (1H‐MRS), and 3) significantly reduced rotational behavior in comparison with control lesioned rats. These observations might be explained by an up‐regulation of growth‐associated protein 43 (GAP‐43) expression because improvements in neurological dysfunction were blocked by injection of MK‐801 in the PBSC‐treated group. In addition, a significant increase in neurotrophic factor expression was found in the ipsilateral hemisphere of the PBSC‐treated group. In summary, this protocol may be a useful strategy for the treatment of clinical PD. © 2009 Wiley‐Liss, Inc.
Vitamin K prevents oxidative cell death by inhibiting activation of 12‐lipoxygenase in developing oligodendrocytes
Authors: Li Jianrong , Wang Hong , Rosenberg Paul A.
Published: February 2009

Oxidative mechanisms of injury are important in many neurological disorders. Developing oligodendrocytes (pre‐OLs) are particularly sensitive to oxidative stress‐mediated injury. We previously demonstrated a novel function of phylloquinone (vitamin K1) and menaquinone 4 (MK‐4; a major form of vitamin K2) in protecting pre‐OLs and immature neurons against glutathione depletion‐induced oxidative damage (Li et al. [ 2003] J. Neurosci. 23:5816–5826). Here we report that vitamin K at nanomolar concentrations prevents arachidonic acid‐induced oxidative injury to pre‐OLs through blocking the activation of 12‐lipoxygenase (12‐LOX). Arachidonic acid metabolism is a potential source for reactive oxygen species (ROS) generation during ischemia and reperfusion. Exposure of pre‐OLs to arachidonic acid resulted in oxidative cell death in a concentration‐dependent manner. Administration of vitamin K (K1 and MK‐4) completely prevented the toxicity. Consistent with our previous findings, inhibitors of 12‐LOX abolished ROS production and cell death, indicating that activation of 12‐LOX is a key event in arachidonic acid‐induced pre‐OL death. Vitamin K1 and MK‐4 significantly blocked 12‐LOX activation and prevented ROS accumulation in pre‐OLs challenged with arachidonic acid. However, vitamin K itself did not directly inhibit 12‐LOX enzymatic activity when assayed with purified 12‐LOX in vitro. These results suggest that vitamin K, or likely its metabolites, acts upstream of activation of 12‐LOX in pre‐OLs. In summary, our data indicate that vitamin K prevents oxidative cell death by blocking activation of 12‐LOX and ROS generation. © 2009 Wiley‐Liss, Inc.
Contribution of a mitochondrial pathway to excitotoxic neuronal necrosis
Authors: Seo Dae‐Won , Lopez‐Meraz Maria‐Leonor , Allen Suni , Wasterlain Claude Guy , Niquet Jerome
Published: February 2009

It is traditionally thought that excitotoxic necrosis is a passive mechanism that does not require the activation of a cell death program. In this study, we examined the contribution of the cytochrome c‐dependent mitochondrial death pathway to excitotoxic neuronal necrosis, induced by exposing cultured cortical neurons to 1 mM glutamate for 6 hr and blocked by the NMDA antagonist, dizocilpine. Glutamate treatment induced early cytochrome c release, followed by activation of caspase‐9 and caspase‐3. Preincubation with the caspase‐9 inhibitor z‐LEHD‐fmk, the caspase‐3 inhibitor z‐DEVD‐fmk, or the specific pan‐caspase inhibitor Q‐VD‐oph decreased the percentage of propidium iodide‐positive neurons (52.5% ± 3.1%, 39.4% ± 3.5%, 44.6% ± 3%, respectively, vs. 65% ± 3% in glutamate + vehicle). EM studies showed mitochondrial release of cytochrome c in neurons in the early stages of necrosis and cleaved caspase‐3 immunoreactivity in morphologically necrotic neurons. These results suggest that an active mechanism contributes to the demise of a subpopulation of excitotoxic necrotic neurons. © 2009 Wiley‐Liss, Inc.
Lipopolysaccharides enhance the action of bradykinin in enteric neurons via secretion of interleukin‐1β from enteric glial cells
Authors: Murakami Matsuka , Ohta Toshio , Ito Shigeo
Published: February 2009

Functional changes of the enteric nervous system have been observed under inflammatory states of inflammatory bowel disease increasing the endotoxin level. The aim of the present study was to determine the effect of lipopolysaccharides (LPS) on myenteric neuron–glia interaction in vitro. We examined the increase of the intracellular Ca2+ concentration ([Ca2+]i) and the release of interleukin‐1β (IL‐1β) or prostaglandin E2 (PGE2) and COX‐2 expression in myenteric plexus cells from the rat intestine induced by LPS. LPS potentiated BK‐induced [Ca2+]i increases in both myenteric neurons and enteric glial cells, which were suppressed by a B1R antagonist. Only in enteric glial cells, a B1R agonist increased [Ca2+]i. The effects of LPS were blocked by pretreatment with an interleukin‐1 receptor antagonist or by reducing the density of enteric glial cells in culture. LPS prompted the release of IL‐1β from enteric glial cells. The augmenting effects of IL‐1β on the BK‐induced neural [Ca2+]i increase and PGE2 release from enteric glial cells were abolished by a phospholipase A2 (PLA2) inhibitor and a COX inhibitor, and partly suppressed by a COX‐2 inhibitor. IL‐1β up‐regulated the COX‐2 expression in enteric glial cells. LPS promotes IL‐1β secretion from enteric glial cells, resulting in augmentation of the neural response to BK through PGE2 release via glial PLA2 and COX‐2. The alteration of the regulatory effect of glial cells may be the cause of the changes in neural function in the enteric nervous system in inflammatory bowel disease. © 2009 Wiley‐Liss, Inc.
Synaptic mRNAs are modulated by learning
Authors: Ferrara Eugenia , Cefaliello Carolina , Eyman Maria , De Stefano Rosanna , Giuditta Antonio , Crispino Marianna
Published: February 2009

We have recently demonstrated that brain plastic events significantly modify synaptic protein synthesis measured by the incorporation of [35S]methionine in brain synaptosomal proteins. Notably, in rats learning a two‐way active avoidance task, the local synthesis of two synaptic proteins was selectively enhanced. Because this effect may be attributed to transcriptional modulation, we used reverse transcriptase–polymerase chain reaction methods to determine the content of discrete synaptosomal mRNAs in rats exposed to the same training protocol. Correlative analyses between behavioral responses and synaptosomal mRNA content showed that GAT‐1 mRNA (a prevalent presynaptic component) correlates with avoidances and escapes in rat cerebellum, while glial fibrillary acid protein mRNA (an astrocytic component) correlates with freezings in cerebellum and cerebral cortex. These observations support the hypothesis that synaptic protein synthesis may be transcriptionally regulated. The cellular origin of synaptic transcripts is briefly discussed, with special regard to those present at large distances from neuron somas. © 2009 Wiley‐Liss, Inc.
An assessment of temperature and precipitation change projections over Italy from recent global and regional climate model simulations
Authors: Coppola Erika , Giorgi Filippo
Published: February 2009

We present an assessment of climate change projections over the Italian peninsula for the 21st century from the CMIP3 global and PRUDENCE regional model experiments. We consider the A2, A1B, B2 and B1 emission scenarios. The climate change signal over Italy varies seasonally, with maximum warming in summer (up to several °C) and minimum in winter, decreased precipitation over the entire peninsula in summer (locally up to −40%) and a dipolar precipitation change pattern in winter (increase to the north and decrease to the south). Inter‐annual variability increases in all seasons for precipitation and in summer for temperature, while it decreases for winter temperature. The seasonal temperature anomaly probability density functions (PDFs) show a shift as well as a broadening and flattening in future climate conditions, especially in summer. This implies larger increases for extreme hot seasons than mean summer temperatures. The seasonal precipitation anomaly PDFs are greatly affected in summer, with a strong increase of very dry seasons. Moreover, seasons with large precipitation amounts tend to increase in future climate conditions, i.e. we find an increase of very dry (drought prone) and very wet (flood prone) seasons. The magnitude of future climate change depends on the emission scenario and the temperature and precipitation change signals show substantial fine‐scale structure in response to the topographical forcing of the Italian major mountain systems. In addition, the change signal is greater than the inter‐model standard deviation for temperature in all seasons and for precipitation in the summer. Finally, the CMIP3 ensemble captures the observed 20th century trends of temperature and precipitation change over northern Italy. A broad agreement between the projections obtained with the CMIP3 and PRUDENCE ensembles is found, which adds robustness to the findings. Copyright © 2009 Royal Meteorological Society
Recent severe heat waves in central Europe: how to view them in a long‐term prospect?
Authors: Kyselý Jan
Published: February 2009

The study examines whether recent occurrences of severe heat waves in central Europe were exceptional in the context of past fluctuations, and estimates their recurrence probabilities under several climate change scenarios. Using data from a network of meteorological stations in the Czech Republic since 1961, it is found that 1994 was the year with the most severe heat waves over majority of the area. The other seasons with enhanced heat wave characteristics were 1992, 2003 and 2006. Analysis of the long‐term temperature series at Prague‐Klementinum reveals that the July 2006 heat wave, covering 33 consecutive days, was the longest and most severe individual heat wave since 1775. Probabilities of long and severe heat waves are estimated from daily temperature series generated by a first‐order autoregressive model with a deterministic component (incorporating a seasonal cycle and a long‐term trend). The model is validated with respect to the simulation of heat waves in present climate (1961–2006) and subsequently run under several assumptions reflecting various rates of summer warming over 2007–2100. The return period of a heat wave reaching or exceeding the length of the 2006 heat wave in Prague is estimated to be around 120 years in 2006. Owing to an increase in mean summer temperatures, probabilities of very long heat waves have already risen by an order of magnitude over the recent 25 years, and are likely to increase by another order of magnitude by around 2040 under the summer warming rate assumed by the mid‐scenario. Even the lower bound scenario yields a considerable decline of return periods associated with intense heat waves. Nevertheless, the most severe recent heat waves appear to be typical rather of a late 21st century than a mid‐21st century climate. Copyright © 2009 Royal Meteorological Society
Validation of the abrupt change in GPCP precipitation in the Congo River Basin
Authors: Yin Xungang , Gruber Arnold
Published: February 2009

The Global Precipitation Climatology Project (GPCP) monthly precipitation exhibits a significant negative trend during 1979–2004 over southern tropical Africa from the Congo River Basin to the east coast. This trend appears as a more than 20% drop beginning in 1992 in the 6‐year and 9‐year averages of the areal mean GPCP satellite‐gauge precipitation, whose magnitude is largely determined by the gauge analyses. This paper's analysis of satellite precipitation estimates, gauge precipitation analyses, and gauge coverage information suggests that the negative precipitation trend is only true in part of southern tropical Africa but the magnitude is much smaller than that calculated from the GPCP. In the eastern portion of the region, the precipitation drop in the GPCP is confirmed by the satellite‐only estimates but the decrease of more than 16% is amplified by a change in gauge coverage. In the western portion of the region, basically the southern Congo River Basin, all gauge dependent products show a negative precipitation trend, which is much larger in the GPCP merged satellite‐gauged data set, but not supported by the satellite‐only precipitation estimates. In this study we conclude that for the Congo River Basin, where both the mean precipitation and its spatial gradient are high, the spurious negative trend detected in the GPCP precipitation is caused by a significant change in local gauge coverage and the methodology used by the GPCP to merge satellite and gauge data during the analysis period. Copyright © 2009 Royal Meteorological Society
Three‐source partitioning of CO2 efflux from maize field soil by 13C natural abundance
Authors: Werth Martin , Kuzyakov Yakov
Published: February 2009

A natural‐13C‐labeling approach—formerly observed under controlled conditions—was tested in the field to partition total soil CO2 efflux into root respiration, rhizomicrobial respiration, and soil organic matter (SOM) decomposition. Different results were expected in the field due to different climate, site, and microbial properties in contrast to the laboratory. Within this isotopic method, maize was planted on soil with C3‐vegetation history and the total CO2 efflux from soil was subdivided by isotopic mass balance. The C4‐derived C in soil microbial biomass was also determined. Additionally, in a root‐exclusion approach, root‐ and SOM‐derived CO2 were determined by the total CO2 effluxes from maize (Zea mays L.) and bare‐fallow plots. In both approaches, maize‐derived CO2 contributed 22% to 35% to the total CO2 efflux during the growth period, which was comparable to other field studies. In our laboratory study, this CO2 fraction was tripled due to different climate, soil, and sampling conditions. In the natural‐13C‐labeling approach, rhizomicrobial respiration was low compared to other studies, which was related to a low amount of C4‐derived microbial biomass. At the end of the growth period, however, 64% root respiration and 36% rhizomicrobial respiration in relation to total root‐derived CO2 were calculated when considering high isotopic fractionations between SOM, microbial biomass, and CO2. This relationship was closer to the 50% : 50% partitioning described in the literature than without fractionation (23% root respiration, 77% rhizomicrobial respiration). Fractionation processes of 13C must be taken into account when calculating CO2 partitioning in soil. Both methods—natural 13C labeling and root exclusion—showed the same partitioning results when 13C isotopic fractionation during microbial respiration was considered and may therefore be used to separate plant‐ and SOM‐derived CO2 sources.
Lichen colonization of an active volcanic environment: a Raman spectroscopic study of extremophile biomolecular protective strategies
Authors: Jorge‐Villar Susana E. , Edwards Howell G. M.
Published: February 2009

The Raman spectra of some sixteen lichen specimens from eight genera growing on basaltic lava and wood substrates on new lava fields on the Kilauea volcano, Kona, Hawaii, have been analysed in therms of the pigments and biomolecules produced in this extreme environment. Chlorophyll and a carotenoid, probably lutein or astaxanthin, are ubiquitous, but otherwise, a range of protective pigments have been identified, including atranorin, usnic acid, gyrophoric acid, parietin and pulvinic acid dilactone. In addition, some species were observed to produce calcium oxalate monohydrate and dihydrate. Clearly, several strategies for survival were being adopted by the lichens to combat radiation insolation, desiccation and high temperatures; in one case, of the same lichen species colonizing wood and lava in the same location, a different survival strategy could be detected related to their substrate, but further long‐term ecological experiments would be indicated to derive more detailed information on this aspect. Copyright © 2009 John Wiley & Sons, Ltd.
Development of a liquid–liquid extraction method for the determination of pyrimethanil, metalaxyl, dichlofluanid and penconazol during the fermentative process of must by GC‐NPD
Authors: Vaquero‐Fernández Luis , Sanz‐Asensio Jesús , Fernández‐Zurbano Purificación , Sainz‐Ramírez Miguel , López‐Alonso Miguel , Epifanio‐Fernández Sara‐Isabel , Martínez‐Soria Mará‐Teresa
Published: February 2009

BACKGROUND: A rapid, selective and sensitive gas chromatographic method is described for the analysis of four fungicides—pyrimethanil, metalaxyl, penconazol and dichlofluanid, widely used in vineyards—in must and wine, as well as samples during alcoholic fermentation. The effect of alcoholic fermentation on pyrimethanil concentration in synthetic and real musts was studied.

RESULTS: The proposed method was based on liquid–liquid extraction followed by gas chromatography with nitrogen–phosphorus detector. n‐Hexane was the most appropriate solvent for extracting the target fungicides in synthetic and real samples. Quality parameters of the proposed method presented good recovery (ca 78.3% for almost all compounds) and precision (between 1.4% and 6.1%), and quantification limits lower than maximum residue limits. However, for dichlofluanid recovery levels of well over 100% were found.

CONCLUSIONS: There was no matrix effect in the analysis of pyrimethanil, metalaxyl or penconazol, but this effect was observed for dichlofluanid in synthetic and real matrices. The application of the fermentative process on pyrimethanil fungicide caused a high decrease in the concentration of this compound, and no product appeared in degradation. The decrease of pyrimethanil in fermented samples was similar in both synthetic and real samples. Copyright © 2009 Society of Chemical Industry
Expression, purification and refolding of recombinant Cry1Ab/Ac obtained in Escherichia coli as inclusion bodies
Authors: Cao Sishuo , Xu Wentao , Luo Yunbo , He Xiaoyun , Guo Xing , Yuan Yanfang , Yuan Changmei , Huang Kunlun
Published: February 2009

BACKGROUND: The Cry toxins are already a useful alternative or supplement to synthetic chemical pesticide application in commercial agriculture and forest management.

RESULTS: The Cry1ab/ac gene from Bacillus thuringiensis was cloned from the genome of genetically modified rice by polymerase chain reaction (PCR). Owing to the large number of Escherichia coli low‐usage codons in the Cry1ab/ac gene, the first 20 codons were optimised by PCR to improve the expression of the Cry1ab/ac gene in E. coli. The Cry1Ab/Ac protein was highly expressed in E. coli as inclusion bodies that could be dissolved in 8 mol L−1 urea and purified on a His Trap™ FF crude column under denaturing conditions. The purified Cry1Ab/Ac protein was dialysed in refolding buffers to obtain a soluble and biologically active protein. To achieve better biological activity, the His‐tag was digested from the Cry1Ab/Ac protein with enterokinase, and the Cry1Ab/Ac protein was further purified by gel filtration on a fast performance liquid chromatography Superdex 75 HR 10/30 column using an AKTA purifier. The identity of the purified Cry1Ab/Ac protein sequence was confirmed by western blot and matrix‐assisted laser desorption ionisation time‐of‐flight mass spectrometry. The final purified Cry1Ab/Ac protein was 99.2% pure and retained its biological activity, as determined in a growth inhibition assay of Chilo suppressalis.

CONCLUSION: The purified Cry1Ab/Ac protein could be used to evaluate the food safety of transgenic plants containing the Cry1ab/ac gene and to produce antibodies for immune‐based methods employed in the detection of genetically modified organisms containing the Cry1ab or Cry1ac gene. It might also serve as a new biological insecticide to reduce the use of broad‐spectrum insecticides. Copyright © 2009 Society of Chemical Industry
Effect of intensifying high‐temperature ripening on proteolysis, lipolysis and flavor of Jinhua ham
Authors: Zhang Jianhao , Zhen Zongyuan , Zhang Wangang , Zeng Tao , Zhou Guanghong
Published: February 2009

BACKGROUND: The flavor quality of dry‐cured ham comes from proteolysis, lipolysis and lipid oxidation, Maillard reaction and Strecker amino acid degradation. Intense proteolysis, lipolysis and lipid oxidation make major contributions to flavor development of dry‐cured ham. Increasing the temperature in fermenting and ripening could promote these reactions and accelerate flavor development in dry‐cured hams. The specific aroma flavor of Jinhua ham is developed only during long‐time high‐temperature ripening in July and August. Our objective was to effectively shorten the process time by intense high‐temperature ripening based on the flavor and quality features of traditional Jinhua ham.

RESULTS: Muscle dehydration rate of 80‐day ripened hams (29.43 ± 1.16%) was higher than that of the traditional process (P < 0.05). The total free fatty acids in ripened hams of 45–80 days were all higher than that of traditional hams (P < 0.05) and the level of TBARS was significantly lower (P < 0.01). The flavor profile of modern‐processed hams was different from that of the traditional Jinhua ham. The contents of carboxylic acids and aldehydes were obviously higher than those of the traditional products (P < 0.05). The results of organoleptic evaluation for flavor and quality showed that 80‐day ripened hams reached the first‐grade level of traditional Jinhua ham.

CONCLUSION: Long‐time (25–30 days) intensifying high‐temperature ripening (35–37 °C) could accelerate the proteolysis, lipolysis, lipids oxidation, flavor development and effectively shorten the process time based on the traditional flavor and quality features of dry‐cured ham. Copyright © 2009 Society of Chemical Industry
Application of polymerase chain reaction–restriction fragment length polymorphism analysis and lab‐on‐a‐chip capillary electrophoresis for the specific identification of game and domestic meats
Authors: Fajardo Violeta , González Isabel , Dooley John , Garret Steve , Brown Helen M , García Teresa , Martín Rosario
Published: February 2009

BACKGROUND: The objective of this study was to adapt and improve previously published polymerase chain reaction–restriction fragment length polymorphism (PCR‐RFLP) methods aimed at the identification of game and domestic meats, by replacing the gel electrophoretic steps for DNA fragment analysis by a chip‐based capillary electrophoresis system.

RESULTS: PCR amplification of a mitochondrial 12S rRNA gene fragment and subsequent digestion of the amplicons with either MseI or a combination of MboII, BslI, and ApoI endonucleases generated characteristic PCR‐RFLP profiles that allowed discrimination among ten relevant game and domestic meat species. The Agilent 2100 Bioanalyzer utilises capillary electrophoresis on a microchip device that is capable of rapidly sizing DNA fragments, offering a valuable recent development for the analysis of complex DNA banding patterns.

CONCLUSION: Results obtained in this work indicated that banding resolution on the system was sensitive, with detection of some small DNA fragments that were not observed with the published conventional PCR‐RFLP gel‐based method. Therefore, the new faster and easy handling procedure provides an additional powerful tool that can be employed for meat speciation. Copyright © 2009 Society of Chemical Industry
Chemical composition and antioxidant and radical‐scavenging activities ofPeriploca laevigataroot bark extracts
Authors: Mohamed Hajji , Ons Masmoudi , Yosra Ellouz‐Triki , Rayda Siala , Neji Gharsallah , Moncef Nasri
Published: February 2009

BACKGROUND:The root powder ofPeriploca laevigatais used for preparing soft drinks and as an aromatic in Tunisia. The infusion or decoction of its root bark has widespread use in folk medicine. The plant is used to treat digestive disorders and hypertensive effects as well as other health problems.

RESULTS:The antioxidant activities of extracts ofP. laevigataroot bark obtained with solvents of different polarity were investigated using assays of 1,1‐diphenyl‐2‐picrylhydrazyl radical‐scavenging activity, ferric‐reducing capacity, β‐carotene‐bleaching ability, hydroxyl radical‐scavenging activity and lipid peroxidation inhibition. The methanol extract, with the highest amount of total phenolics and flavonoids, showed the highest antioxidant activities in all assays, followed by the water extract. Gas chromatography/mass spectrometry was used to determine the composition of the water and methanol extracts. Thirty‐four compounds were identified in the methanol extract, with proflavine (516.2 g kg−1dry matter (DM)) and 4‐methoxysalicylaldehyde (198.3 g kg−1DM) being the most abundant. Sixteen compounds were identified in the water extract, of which 4‐hydroxy‐3‐methoxyphenethylene glycol (351.2 g kg−1DM) was the main component.

CONCLUSION:As far as is known, this is the first report on the chemical composition and biological activities of phenolic extracts fromP. laevigata. The results of the study indicate that the root bark of this plant might be a good candidate for further investigation in developing new antioxidants. Copyright © 2009 Society of Chemical Industry