bluuebunny/crossref_metadata_2025 huggingface dataset:
Soluble CD14 levels are increased and inversely correlated with the levels of hepatitis B surface antigen in chronic hepatitis B patients
Authors: Steyaert Sophia , Vanlandschoot Peter , Van Vlierberghe Hans , Diepolder Helmut , Leroux‐Roels Geert
Published: August 2003

Because it was observed recently that yeast‐derived recombinant HBsAg interacts in a lipopolysaccharide binding protein‐dependent manner with CD14 expressed on human monocytes, we investigated whether HBsAg influences the serum levels of sCD14, lipopolysaccharide binding protein and C‐reactive protein in hepatitis B patients. Samples from acute and chronic hepatitis B and chronic hepatitis C patients were tested. All analytes were measured using commercial assays. HBsAg was quantified using an NIBSC titrated standard. sCD14 levels were higher in chronic hepatitis B and C patients than in healthy controls (P = 0.0006 and P < 0.0001, respectively). In chronic hepatitis B patients an inverse correlation was found between sCD14 and HBsAg (P = 0.0309). Lipopolysaccharide binding protein and C‐reactive protein levels were higher in acute hepatitis B patients than in control subjects (P = 0.0217 and P = 0.0034, respectively). In chronic hepatitis B and C, sCD14 and C‐reactive protein levels were higher in cirrhotic than in non‐cirrhotic patients (P = 0.0072 and P = 0.0223, respectively). J. Med. Virol. 71:188–194, 2003. © 2003 Wiley‐Liss, Inc.
Characterization and evolution of NS5A quasispecies of hepatitis C virus genotype 1b in patients with different stages of liver disease
Authors: Franco Sandra , Giménez‐Barcons Mireia , Puig‐Basagoiti Francesc , Furcic Ivana , Sánchez‐Tapias José‐María , Rodés Juan , Sáiz Juan‐Carlos
Published: August 2003

The quasispecies nature of hepatitis C virus (HCV) is thought to play a central role in modulating viral functions. Recent work has linked NS5A protein with viral replication, resistance to interferon (IFN), and control of cellular growth, probably through the interaction of its protein kinase R (double stranded RNA‐activated protein kinase, PKR) binding domain (PKR‐bd) with cellular PKR, but knowledge of how PKR‐bd viral population evolves during disease progression is limited. Since we have previously described an association between amino acid composition of the PKR‐bd and the presence of HCC, in this report we further investigated the dynamic behavior of viral population parameters by sequencing an average of 20 clones per sample in 27 samples from 19 untreated patients with different degrees of liver disease, 8 of whom were followed over time. Viral population parameters varied widely from patient to patient, but no differences were observed in the complexity, diversity, types of nucleotide changes, or evolutionary pattern of the quasispecies according to the stage of liver disease. In five samples, we detected “quasispecies‐tails”; that is, clones whose minimum genetic distance to the remaining clones of their own quasispecies were higher than the maximum genetic distance found between any other two clones of the same sample. In summary, independent of the degree of liver disease, or the mutations detected within the consensus sequence of the PKR‐bd, the NS5A of HCV presents a flexible and variable quasispecies structure that remains largely stable during the natural course of an HCV infection, highlighting the central role of NS5A protein in viral life cycle. J. Med. Virol. 71:195–204, 2003. © 2003 Wiley‐Liss, Inc.
Loss of hepatitis C virus RNA after parturition in female patients with chronic HCV infection
Authors: Hattori Yuko , Orito Etsuro , Ohno Tomoyoshi , Sugauchi Fuminaka , Suzuki Seiji , Sugiura Mayumi , Suzumori Kaoru , Hattori Kohei , Ueda Ryuzo , Mizokami Masashi
Published: August 2003

It is unclear whether pregnancy has any influence on chronic hepatitis C virus (HCV) infection. The aim of this study was to investigate the relationship between pregnancy and parturition with HCV viremia levels and the natural resolution of HCV RNA. Twenty‐two pregnant patients and 120 nonpregnant control female patients, both positive for anti‐HCV and HCV RNA, were studied. The HCV core protein levels were quantified by enzyme immunoassay, and HCV RNA was detected by reverse transcription‐polymerase chain reaction (RT‐PCR). Of the pregnant group, two females whose HCV RNA was negative continuously for more than 6 months lost HCV RNA permanently after parturition, and one female whose level of HCV core protein was intermittently under the limit of detection level lost HCV RNA intermittently. In the control group, only one female lost HCV RNA persistently, and one lost HCV RNA intermittently after she developed liver cirrhosis. At 3 months after parturition, the HCV core protein level was <15 fmol/L in all patients who lost HCV RNA, while the HCV core protein level was ≥15 fmol/L in 81.3% of the patients who persistently had HCV RNA (P = 0.03). Significantly more pregnant patients lost HCV RNA than did nonpregnant controls. These findings suggest that pregnancy and parturition appear to influence the clinical course of HCV infection. J. Med. Virol. 71:205–211, 2003. © 2003 Wiley‐Liss, Inc.
Polymorphisms in tumour necrosis factor‐α, transforming growth factor‐β, interleukin‐10, interleukin‐6, interferon‐γ, and outcome of hepatitis C virus infection
Authors: Barrett S. , Collins M. , Kenny C. , Ryan E. , Keane C.O. , Crowe J.
Published: August 2003

Cytokines play a key role in the regulation of immune responses. In hepatitis C virus infection (HCV), the production of inappropriate cytokine levels appears to contribute to viral persistence and to affect response to therapy. Cytokine genes are polymorphic at specific sites, and certain mutations located within coding/regulatory regions have been shown to affect the overall expression and secretion of cytokines. The aim of this study was to investigate the frequency of genotypes associated with polymorphisms of TNF‐α, TGF‐β, IL‐10, IL‐6, and IFN‐γ and to determine their association with the outcome of HCV infection. Genotyping was carried out by polymerase chain reaction sequence‐specific primers on genomic DNA isolated from 158 individuals. Of these, 66 had spontaneously recovered from infection (persistently HCV RNA negative), while 92 had persistent infection (persistently HCV RNA positive). All patients were genotyped as high or low producers of TNF‐α and IL‐6 and high, intermediate, or low producers of TGF‐β, IL‐10, and IFN‐γ based on single nucleotide substitutions. A significant proportion of patients with viral clearance were genotyped with a low IL‐6 production profile, whereas those with persistent infection were genotyped with a high production profile (P = 0.02). No associations were observed between polymorphisms of TNF‐α, IL‐10, or IFN‐γ and viral clearance or persistent infection. Furthermore, there were no associations between cytokine genotypes and severity of disease. Inheritance of some genotypes associated with polymorphisms of cytokine genes, such as IL‐6, may be host genetic factors associated with outcome of HCV in a well‐defined ethnically homogeneous cohort. J. Med. Virol. 71:212–218, 2003. © 2003 Wiley‐Liss, Inc.
High rate of hepatitis C virus infection and predominance of genotype 4 among elderly inhabitants of a remote village of the rain forest of South Cameroon
Authors: Njouom Richard , Pasquier Christophe , Ayouba Ahidjo , Gessain Antoine , Froment Alain , Mfoupouendoun Jermie , Pouillot Regis , Dubois Martine , Sandres‐Sauné Karine , Thonnon Jocelyn , Izopet Jacques , Nerrienet Eric
Published: August 2003

To determine the prevalence of hepatitis C virus (HCV) infection and genotype distribution in a rural isolated area of Central Africa, plasma of 409 Bantous (mean age, 32 years; range, 2–78 years) living in a remote village of the rain forest of Southern Cameroon was screened for anti‐HCV antibodies. HCV seropositive samples were also subjected to qualitative detection of viral RNA. HCV antibodies were detected in 70 (17.1%) individuals, 48 (68.8%) of whom had detectable viremia. The seroprevalence did not differ by gender (P = 0.37), but increased significantly with age (P < 0.05), with a strong increase in the oldest age groups. Indeed, nearly one‐half (48%) of the adults >50 years old were HCV seropositive. The characterization of the viral genotypes indicated that most of the HCV strains were of genotype 4 (76%), while genotype 2 (16%), and 1 (8%) were marginally represented. The results suggest a cohort effect with an old, possibly iatrogenic, group exposure rather than a continuous exposure. A more in‐depth population‐based epidemiological study is needed to address this issue further. J. Med. Virol. 71:219–225, 2003. © 2003 Wiley‐Liss, Inc.
Genotyping of GB virus C by restriction pattern analysis of the 5′ untranslated region
Authors: Schleicher Sabine B. , Flehmig Bertram F.
Published: August 2003

GB virus C (GBV‐C) is a virus that has been proposed as a member of the Flaviviridae family, distantly related to hepatitis C virus (HCV). The virus is able to infect humans parenterally and perinatally, although its true pathogenicity remains unknown. The 5′ terminal region of GBV‐C is the most highly conserved region of the virus genome. Comparison of 5′ untranslated region (5′ UTR) sequences from GBV‐C infected individuals shows that variation is limited to particular sites that are often covariant and associated with different virus genotypes. Extensive sequence analysis of the GBV‐C genome provides evidence for the existence of at least five major genotypes, some of which can be further divided into subtypes. For genotyping by restriction fragment length polymorphism (RFLP), it is essential to identify genomic positions that not only reflect genotype differences, but that also harbor restriction sites that allow recognition of these differences. Restriction site analysis of type‐specific sequence motifs predicted that endonucleases BsmFI, HaeII, HinfI, and ScrFI could be used for the identification all known genotypes (types 1–5) with 99.6% accuracy. The method was applied to serum samples from 46 chronic GBV‐C carriers of heterogeneous geographical and ethnic origin, comparing observed cleavage patterns of GBV‐C variants amplified by reverse transcriptase‐polymerase chain reaction (RT‐PCR) of the 5′ UTR with the RFLP predicted from sequences deposited in GenBank database. cDNA sequencing and subsequent alignment of the 46 GBV‐C isolates confirmed RFLP profiles predicted theoretically. The observed geographical distribution of genotypes is also in agreement with previous reports. This method may be useful for rapid and reliable characterization of GBV‐C isolates when either epidemiological or transmission studies are carried out. J. Med. Virol. 71:226–232, 2003. © 2003 Wiley‐Liss, Inc.
Acute inflammatory response and remodeling of airway epithelium after subspecies B1 human adenovirus infection of the mouse lower respiratory tract
Authors: Kajon Adriana E. , Gigliotti Andrew P. , Harrod Kevin S.
Published: August 2003

Despite the well‐recognized role of adenoviruses of species B in the etiology of severe respiratory disease, the lack of an experimental in vivo model system has limited the understanding of the molecular pathogenesis of species B adenovirus‐induced pneumonia. Intratracheal instillation of 5 × 108 plaque‐forming units (pfu) of adenoviruses 3p and 7h resulted in a robust inflammatory response in the lungs of infected mice. A marked infiltration of neutrophils into the lung air spaces was observed at 1 and 2 days postinfection (dpi), with a concomitant increase in the levels of neutrophil chemokines MIP‐2 and KC. The overall histological severity scores were significantly higher for Ad3p‐infected mice at 2 dpi, but similar between the two viruses at other time points. Remodeling of the airway epithelia and mucous cell metaplasia were noted in the proximal airways of infected mice, indicating marked epithelial differentiation and/or injury. The proinflammatory cytokines interleukin‐β (IL‐1β), tumor necrosis factor‐α (TNF‐α), interferon‐γ (IFN‐γ• •), and interleukin‐12 (IL‐12) were induced by viral infection. Expression of the early viral immunomodulatory genes E3‐15.3K and E3gp19K mRNA was readily detectable in the lungs of infected mice by reverse transcription‐polymerase chain reaction (RT‐PCR) at 1 and 2 dpi, coinciding with the peak levels of TNF‐α. While the detection of gp19K mRNA declined thereafter, 15.3K mRNA was detectable up to 6 dpi. Our results indicate that human Ad3 and Ad7 cause marked pulmonary pathology, inducing similar host responses in the respiratory tract, thus validating the use of the mouse model for the study of early virus‐host interactions during lung infection by adenoviruses of subspecies B1. J. Med. Virol. 71:233–244, 2003. © 2003 Wiley‐Liss, Inc.
Persistent gastroenteritis in children infected with astrovirus: Association with serotype‐3 strains
Authors: Caballero Santiago , Guix Susana , El‐Senousy Waled Morsy , Calicó Ignasi , Pintó Rosa M. , Bosch Albert
Published: August 2003

The relationship between cases of persistent diarrhoea and the levels and type of human astrovirus was investigated. The potential correlation between human astrovirus excretion levels and the occurrence of protracted gastroenteritis was elucidated after quantifying astroviruses in faecal samples by a competitive RT‐PCR. This assay was developed employing an internal RNA standard constructed for this purpose and showed a threshold of positivity of 3.4 × 104 genomes per gram of faeces. By this procedure, the levels of astrovirus, belonging to serotypes 1, 2, 3, 4, and 8, in faecal samples could be ascertained to range from 3.4 × 108 to 1 × 1013 per gram of faeces. The mean viral titre in the serotype 3‐containing faeces was higher than in any of the other serotype‐containing samples. In children with no background disease, persistent gastroenteritis cases were detected in 8.5% of the astrovirus infections, and 37.5% of those were associated with astrovirus type 3 infection. In addition, 42.9% of astrovirus 3 isolates were implicated with persistent cases, some of them lasting for 3 months. Other type 3 isolates, detected in the faeces in very large numbers, caused severe gastroenteritis. J. Med. Virol. 71:245–250, 2003. © 2003 Wiley‐Liss, Inc.
Flow cytometric analysis of cytomegalovirus‐specific cell‐mediated immunity in the congenital infection
Authors: Hayashi Naomi , Kimura Hiroshi , Morishima Tsuneo , Tanaka Naoko , Tsurumi Tatsuya , Kuzushima Kiyotaka
Published: August 2003

Flow cytometric analysis of CD4+ and CD8+ T cells specific to human cytomegalovirus (CMV) was undertaken in seven patients with congenital CMV infection, six healthy infants who had acquired infection, and six CMV‐seropositive adults. Intracellular cytokine assays showed that 0.03–2.23% of CD4+ T cells in the healthy infants and adults produced interferon‐γ (IFN‐γ) in response to CMV antigens. In contrast, such CD4+ T cells were almost undetectable in patients with congenital CMV infection who were younger than 2 years of age. Tetrameric major histocompatibility complex/peptide complex analysis demonstrated HLA*A2402‐restricted phosphoprotein 65‐specific CD8+ T cells to be present in most healthy infants and adults tested, but almost absent in the patients. Interestingly, CMV‐specific CD4+ T‐cell responses were observed in two patients with congenital infection beyond the age of 5 years. The present study points to impairment of CMV‐specific cellular immunity in patients in single‐cell levels with congenital CMV infection during the infant period and possible restoration in later childhood. J. Med. Virol. 71:251–258, 2003. © 2003 Wiley‐Liss, Inc.
Correlation between increased platelet‐associated IgG and thrombocytopenia in secondary dengue virus infections
Authors: Oishi Kazunori , Inoue Shingo , Cinco Maria T.D.D. , Dimaano Efren M. , Alera Maria T.P. , Alfon Jhoe A.R. , Abanes Ferdinand , Cruz Deu J.M. , Matias Ronald R. , Matsuura Hiromi , Hasebe Futoshi , Tanimura Susumu , Kumatori Atsushi , Morita Kouichi , Natividad Filipinas F. , Nagatake Tsuyoshi
Published: August 2003

Although the public health impact of dengue is increasing rapidly, the mechanism of thrombocytopenia in this disease remains unknown. To elucidate this mechanism, the relationship between platelet‐associated IgG (PAIgG) and platelet count in 53 patients in the acute phase of secondary dengue virus infection was investigated in a prospective‐hospital‐based study. A significant inverse correlation between the two parameters was found in these patients, while no correlation was observed in healthy volunteers. The low baseline platelet counts during the acute phase in 12 patients with secondary dengue virus infection significantly increased during the convalescent phase, while the increased PAIgG levels during the acute phase in these patients significantly decreased during the convalescent phase. Anti‐platelet IgG autoantibody was detected rarely in the plasma of 53 patients with secondary dengue infection. The involvement of anti‐dengue virus IgG was also shown in platelets from all of 8 patients in the acute phase of secondary dengue virus infection. These findings suggest that PAIgG formation involving anti‐dengue virus IgG plays a pivotal role in the induction of transient thrombocytopenia during the acute phase of secondary dengue virus infection. J. Med. Virol. 71:259–264, 2003. © 2003 Wiley‐Liss, Inc.
High prevalence of HPV 16 in South African women with cancer of the cervix and cervical intraepithelial neoplasia
Authors: Kay Patti , Soeters Robbert , Nevin James , Denny Lynette , Dehaeck Catherine M.C. , Williamson Anna‐Lise
Published: August 2003

Despite the high prevalence of cervical cancer and cervical neoplasias in South Africa, few studies have been performed in this region to establish which human papillomavirus (HPV) types are associated with the development of high‐grade cervical intraepithelial neoplasia lesions and cervical cancer. To investigate these prevalence rates, punch biopsies were obtained from 56 women with cervical cancer and 141 women with histologically diagnosed cervical intraepithelial neoplasia 2 or 3 lesions. Nested polymerase chain reaction (PCR) using consensus degenerate PCR primers was performed for the detection of HPV DNA and HPV typing was done by restriction fragment length polymorphism. Forty‐seven (94%) of the cervical cancer and 114 (88%) of the cervical intraepithelial neoplasia 2/3 biopsies were positive for HPV DNA. The prevalence rates of the HPV types detected in the cervical cancer biopsies were HPV 16 (82%), HPV 18, (10%), HPV 33 (10%), HPV 31 (2%), HPV 58 (2%), HPV 35 (2%), and HPV 59 (2%). The cervical intraepithelial neoplasia lesions contained HPV 16 (56.6%), HPV 33 (14%), HPV 31 (10.9%), HPV X (7%), HPV 52 (3.9), HPV 58 (3.1%), HPV 35 (2.3%), HPV 18 (1.6%), HPV 11 (0.8%). Five of the nine fragments that were not typed by the RFLP, designated HPV‐X, were sequenced to give HPV6 (1/5), HPV 26 (2/5), HPV 68 (1/5), and candHPV 87 (1/5). HPV 58 was detected in one cervical cancer biopsy and four biopsies from cervical intraepithelial neoplasia grade 3 lesions and was shown to be a previously described variant [Williamson and Rybicki (1991) J. Med. Virol. 33:165–171]. In addition, a cervical intraepithelial neoplasia grade 2 lesion was shown to harbour HPV type HAN2294 (cand HPV 87). The results of this study indicate that cervical cancer and cervical intraepithelial neoplasia 2/3 are largely associated with HPV 16 infection in this group of South African women and, therefore, an effective HPV 16 based vaccine should prevent the development of cervical cancer in a large proportion of women from this region of South Africa. J. Med. Virol. 71:265–273, 2003. © 2003 Wiley‐Liss, Inc.
Serotype‐specific mucosal immune response and subsequent poliovirus replication in vaccinated children
Authors: Samoilovich Elena , Roivainen Merja , Titov L.P. , Hovi Tapani
Published: August 2003

A total of 32 unimmunized children (median age 5 months) living in an orphanage in Minsk, Belarus, were vaccinated with three doses of oral poliovirus vaccine (OPV) with a 60‐day interval between the doses. Blood samples were drawn before the immunizations and 45–50 days after each vaccine dose. Excretion of the vaccine viruses was followed by examining fecal specimens collected weekly after each vaccine dose. All children seroconverted by the second dose of OPV at the latest to all three serotypes of poliovirus but differences were seen regarding the intestinal responses. The strongest responses in both neutralizing antibodies and virus‐binding IgA in fecal suspensions were seen toward poliovirus Type 2. Consistent with this, relatively little poliovirus Type 2 excretion was seen after the second and third doses of OPV as compared to that of poliovirus Type 1 or Type 3. The delayed development of functional intestinal immunity against the latter serotypes was associated with relatively weaker intestinal antibody responses compared to poliovirus Type 2. In the case of poliovirus Type 3, about 10% of children were still excreting the vaccine virus 9 weeks after administering the third dose. These results are consistent with epidemiological observations of the serotype‐specific efficacy of OPV immunizations. The proportion of specimens showing nonpolio enteric viruses gradually increased through the 6‐month study period. This may reflect the possibility that after the first dose of OPV, intensive replication of the vaccine viruses may out compete the nonpolio viruses in the intestines of the vaccinees or, alternatively, mask nonpolio viruses during the cell culture isolation procedure. J. Med. Virol. 71:274–280, 2003. © 2003 Wiley‐Liss, Inc.
Case report: Isolation of a European bat lyssavirus type 2a from a fatal human case of rabies encephalitis
Authors: Fooks Anthony R. , McElhinney Lorraine M. , Pounder Derrick J. , Finnegan Christopher J. , Mansfield Karen , Johnson Nicholas , Brookes Sharon M. , Parsons Graham , White Kathleen , McIntyre Paul G. , Nathwani Dilip
Published: August 2003

A 55‐year‐old bat conservationist was admitted to Ninewells Hospital, Dundee, Scotland, on November 11, 2002, with an acute haematemesis. He gave a 5‐day history of pain and paraesthesia in the left arm, followed by increasing weakness of his limbs with evidence of an evolving encephalitis with cerebellar involvement. The patient had never been vaccinated against rabies and did not receive postexposure treatment. Using a hemi‐nested reverse transcriptase‐polymerase chain reaction (RT‐PCR), saliva samples taken intravitam from different dates proved positive for rabies. A 400‐bp region of the nucleoprotein gene was sequenced for confirmation and identified a strain of European bat lyssavirus (EBLV) type 2a. The diagnosis was confirmed using the fluorescent antibody test (FAT) and by RT‐PCR on three brain samples (cerebellum, medulla, and hippocampus) taken at autopsy. In addition, a mouse inoculation test (MIT) was performed. Between 13 and 17 days postinfection, clinical signs of a rabies‐like illness had developed in all five inoculated mice. Brain smears from each infected animal were positive by the FAT and viable virus was isolated. This fatal incident is only the second confirmed case of an EBLV type‐2 infection in a human after exposure to bats. J. Med. Virol. 71:281–289, 2003. Copyright © Crown Copyright 2003. Recorded with the permission of the controller of the Majesty's Stationery Office. Published by Wiley‐Liss, Inc.
RSV‐induced bronchiolitis but not upper respiratory tract infection is accompanied by an increased nasal IL‐18 response
Authors: van Benten Inesz J. , van Drunen Cornelis M. , Koopman Laurens P. , KleinJan Alex , van Middelkoop Barbara C. , de Waal Leon , Osterhaus Albert D.M.E. , Neijens Herman J. , Fokkens Wytske J.
Published: August 2003

The aim of this study was to investigate potential differences in the local nasal immune response between bronchiolitis and upper respiratory tract infection induced by respiratory syncytial virus (RSV). Nasal brush samples were obtained from 14 infants with RSV bronchiolitis and from 8 infants with RSV upper respiratory tract infection. The samples were taken during infection (acute phase) and 2–4 weeks later (convalescent phase). Cytospin preparations were stained immunohistochemically for T cells, macrophages, and eosinophils. Staining also took place for intercellular adhesion molecule‐1 (ICAM‐1), T‐helper 1 (Th1)‐like (interleukin‐12 [IL‐12], interferon‐γ [IFN‐γ]), Th2‐like (IL‐4, IL‐10), and proinflammatory cytokines (IL‐6, IL‐8, IL‐18). During both RSV‐induced bronchiolitis and upper respiratory tract infection, cellular inflammation was observed. This was characterised by an increase in the numbers of nasal macrophages, which tended to be higher in bronchiolitis than in upper respiratory tract infection. Numbers of T lymphocytes and ICAM‐1 positive cells increased during both bronchiolitis and upper respiratory tract infection. There were no differences between numbers in the groups. Interestingly, a distinct nasal proinflammatory cytokine response was observed in RSV‐induced bronchiolitis. This is characterised by an increase in the number of IL‐18 positive cells. This increase is specific for bronchiolitis, as a similar increase could not be detected in RSV‐induced upper respiratory tract infection. Numbers of IL‐6 and IL‐12 positive cells were higher in both bronchiolitis and upper respiratory tract infection, and there were no differences between the groups. By contrast, the number of IL‐8, IFN‐γ, IL‐4, and IL‐10‐positive cells remained constant. In conclusion, clear differences were found in nasal immune responses of children with RSV‐induced upper respiratory tract infection or bronchiolitis. The induction of a strong IL‐18 response was typical for bronchiolitis, as this could not be observed in RSV‐induced upper respiratory tract infection, and could explain the eosinophilia that is observed frequently during bronchiolitis. J. Med. Virol. 71:290–297, 2003. © 2003 Wiley‐Liss, Inc.
Prevalence of anti‐human respiratory syncytial virus antibodies over three consecutive years in a healthy adult population
Authors: Chala Silvia R. , García‐Barreno Blanca , Melero José A. , Palomo Concepción
Published: August 2003

The titers of anti‐human respiratory syncytial virus (HRSV) antibodies have been measured by enzyme‐linked immunosorbent assay (ELISA) in a cohort of healthy adult volunteers over 3 consecutive years. Significant increases in the level of these antibodies were detected in a small percentage of individuals (3–3.3%), when paired serum samples taken from two consecutive seasons were compared. In these cases, a significant increase was generally noted in the level of antibodies directed against either the F or the G glycoproteins, and in the level of neutralizing antibodies. However, no correlation was found between the level of binding and neutralizing antibodies, when samples representative of the population under study were analyzed. These results indicate some level of infection by HRSV in the adult population. The implications of these results for the epidemiology of HRSV are discussed. J. Med. Virol. 71:298–304, 2003. © 2003 Wiley‐Liss, Inc.
Antigenic and genetic variability of human respiratory syncytial viruses (group A) isolated in Uruguay and Argentina: 1993–2001
Authors: Frabasile Sandra , Delfraro Adriana , Facal Luján , Videla Cristina , Galiano Mónica , de Sierra María José , Ruchansky Dora , Vitureira Nathalia , Berois Mabel , Carballal Guadalupe , Russi José , Arbiza Juan
Published: August 2003

The antigenic and genetic diversity of G glycoprotein from 25 human respiratory viruses (group A) isolated during nine consecutive epidemics (1993–2001) in Montevideo, Uruguay, and 7 strains isolated in Buenos Aires, Argentina, in the same period were analyzed. Genetic variability was evaluated by partial sequence of the G protein gene. Phylogenetic analysis indicated that most Uruguayan and Argentinean group A isolates clustered into three genotypes: GA5, GA2, and GA1. Some strains clustered into the GA3 genotype characterized previously. The antigenic analysis was carried out with a panel of anti‐G monoclonal antibodies that recognized conserved and strain‐specific epitopes. A close correlation between the antigenic and genetic relatedness of the strains analyzed was observed. J. Med. Virol. 71:305–312, 2003. © 2003 Wiley‐Liss, Inc.
Molecular characterisation of varicella‐zoster virus strains in Germany and differentiation from the Oka vaccine strain
Authors: Sauerbrei A. , Eichhorn U. , Gawellek S. , Egerer R. , Schacke M. , Wutzler P.
Published: August 2003

With the introduction of varicella vaccination, surveillance of varicella‐zoster virus (VZV) strains occurring in cases of chickenpox or zoster should be considered. Differentiating Oka vaccine strain from wild‐type VZV can be achieved only using molecular genotyping. In the present study, the VZV genotype was examined in 53 VZV strains isolated from patients with varicella or zoster and in 73 samples from skin eruptions, cerebrospinal fluid, and throat swabs obtained from patients with VZV infections in Germany. The polymerase chain reaction and restriction fragment length polymorphisms analysis using DNA fragments of the open reading frames 38, 54, 62, and the R5 repeat region were used. Whereas all VZV isolates could be typed, direct genotyping of viral DNA in patients' samples was achieved in 63 of 73 cases (86.3%). The dominant genotype of VZV found in 88.8% of 116 patients had the wild‐type pattern PstI+ BglI− R5A followed by the wild‐genotype PstI+ BglI+ R5A in 6.0%, the wild‐genotype PstI+ BglI− R5B in 3.4%, the wild‐genotype PstI+ BglI− R5C and the Oka vaccine genotype PstI− BglI+ R5B in 0.9% of patients each. BglI− wild‐types were found in 90.7% of patients with zoster and in 9.3% of patients with varicella. By contrast, the BglI+ wild‐type was diagnosed in five patients with varicella and in two patients with zoster. In conclusion, VZV strains found in Germany are similar to strains circulating in the United States and the United Kingdom. VZV wild‐type strains containing a BglI restriction site in ORF 54 as well as Oka vaccine strains can rarely be detected. J. Med. Virol. 71:313–319, 2003. © 2003 Wiley‐Liss, Inc.
Early NFκB activation is inhibited during focal cerebral ischemia in interleukin‐1β‐converting enzyme deficient mice
Authors: Huang Feng‐Peng , Wang Zhi‐Qiu , Wu Du‐Chu , Schielke Gerald P. , Sun Yi , Yang Guo‐Yuan
Published: August 2003

Our previous study demonstrated that the inhibition of interleukin‐1β (IL‐1β) reduces ischemic brain injury; however, the molecular mechanism of the action of IL‐1 in cerebral ischemia is unclear. We are investigating currently the role of NFκB during focal cerebral ischemia, using mutant mice deficient in the interleukin‐1 converting enzyme gene (ICE KO) in a middle cerebral artery occlusion (MCAO) model. Adult male ICE KO and wild‐type mice (n = 120) underwent up to 24 hr of permanent MCAO. Cytoplasmic phospho‐NFκB/p65 expression in ischemic brain was examined using Western blot analysis and immunohistochemistry. NFκB DNA‐binding activity was detected using electrophoretic mobility shift assay (EMSA). Furthermore, ICAM‐1 expression was examined in both the ICE KO and wild‐type mice (WT). Western blot analysis and immunostaining showed that the level of cytosolic phosphorylated NFκB/p65 increased after 2 and 4 hr of MCAO in WT mice; however, NFκB/p65 was significantly reduced after MCAO in the ICE KO mice (P < 0.05). EMSA showed that NFκB DNA‐binding activity increased after MCAO in WT mice; but this effect was reduced in the ICE KO mice. The number of ICAM‐1‐positive vessels in the ischemic hemisphere was greatly attenuated in the ICE KO mice (P < 0.05), which paralleled the results of immunohistochemistry. Our results demonstrate that NFκB phosphorylation is reduced in ICE KO mice, suggesting that ICE or IL‐1 are involved in early NFκB phosphorylation. Because cerebral ischemia induced infarction is significantly reduced in ICE KO mice, we conclude that early NFκB phosphorylation plays a disruptive role in the ischemic process. © 2003 Wiley‐Liss, Inc.
Characterization of the mouse olfactory glutathione S‐transferases during the acute phase response
Authors: Weech Michelle , Quash Michelle , Walters Eric
Published: August 2003

The acute phase response (APR) has been shown to alter expression and activity of biotransformation enzymes, such as the phase I cytochromes P450 and phase II glutathione S‐transferases (GSTs). The cytochromes P450 and GSTs are expressed abundantly and colocalized to non‐neuronal cells of the olfactory mucosa. Previous studies indicate that olfactory cytochromes P450 expression and activity is altered during periods of localized inflammation and infection. Little is understood, however, about the influence of the APR on olfactory GST enzymes. This study investigated effects of the APR on olfactory GST isozymes expression and activity in mouse olfactory mucosa after 24‐hr treatment with the acute phase inducer, polyinosinic: polycytidylic acid (polyIC). Western blot analysis using antibodies directed against specific GST isoforms α (A1‐1), μ (M1‐1), and π (P1‐1) demonstrated that their expression was unaltered by polyIC treatment. In contrast, olfactory P450 2E1 expression was significantly decreased. Enzymatic activity of the olfactory GSTs toward the general substrate, 1‐chloro‐2,4‐dinitrobenzene (CDNB) was unchanged during the APR. Analysis of olfactory glutathione content during the APR showed that it was also unaffected by polyIC. The insensitivity of these olfactory GST isoforms during the APR may play a significant role toward limiting the impact of infection and inflammation on the olfactory system. © 2003 Wiley‐Liss, Inc.
Chronic nicotine administration increases NGF‐like immunoreactivity in frontoparietal cerebral cortex
Authors: Martínez‐Rodríguez R. , Toledano A. , Álvarez M.I. , Turégano L. , Colman O. , Rosés P. , Gómez de Segura I. , De Miguel E.
Published: August 2003

Nicotine/nicotine agonists, which have been proposed as therapeutic agents for the treatment of Alzheimer's disease and other neurodegenerative disorders, produce a wide variety of effects on the nervous system. Some mechanisms involved remain poorly understood. In this work, immunohistochemical techniques were used to determine the effect of nicotine on nerve growth factor (NGF) in the frontoparietal (motor, somatosensory) brain cortex of the albino rat. Nicotine was chronically administered intraperitoneally using osmotic pumps (0.35 mg nicotine base/kg body weight/day for 14 days). An increase in the number and the immunoreaction intensity of NGF‐like positive pyramidal and nonpyramidal neurons of these cortical areas was observed after treatment. Immunopositive astroglial cells were always seen in sections of treated animals but not in controls. The neuropil of control animals was, in general, devoid of reaction, but in treated animals, immunopositive prolongations were located randomly, some in close association with capillaries. At the electron microscopic level, these prolongations were demonstrated as belonging to neurons (dendrites and axons) and astroglial cells. Nicotinic activation of selected neurons and glial cells seems to trigger NGF/neurotrophic mechanisms, suggesting their use may be of benefit in prevention and treatment of neurodegenerative diseases. © 2003 Wiley‐Liss, Inc.
CNS gene expression pattern associated with spontaneous experimental autoimmune encephalomyelitis
Authors: Matejuk Agata , Hopke Corwyn , Dwyer Jami , Subramanian Sandhya , Jones Richard E. , Bourdette Dennis N. , Vandenbark Arthur A. , Offner Halina
Published: August 2003

Transgenic mice with T‐cell receptor (TCR) specific for myelin basic protein (MBP)‐Ac1‐11 peptide and homozygous for the RAG‐1 mutation (T/R− mice) spontaneously develop acute progressive experimental autoimmune encephalomyelitis (Sp‐EAE) mediated by CD4+ T cells. Microarray analysis of spinal cord tissue obtained from symptomatic versus non‐symptomatic T/R− mice revealed strongly upregulated transcripts for genes involved in antigen presentation and processing, signal transduction, transcription regulation, metabolism, development, cell cycle, and many other processes involved in the induction of clinical and pathological signs of Sp‐EAE. Several highly expressed genes were related directly to inflammation, including cytokines/receptors, chemokines/receptors, acute phase, complement molecules, and others. Many CNS‐specific genes were also upregulated in sick mice. Abundance of message for the Tg TCR BV8S2 gene as well as several monocyte/macrophage‐associated genes would suggest that both components play a crucial role in the pathogenesis of Sp‐EAE. The profile of transcriptional changes found during the development of Sp‐EAE provides the first description of the encephalitogenic process in the absence of purposeful immunization with myelin peptides and immune‐enhancing adjuvants. This unique approach is the first to implicate molecules and pathways that contribute naturally to onset of paralysis and demyelination, and thus may provide unique insights and novel treatment strategies for human diseases such as multiple sclerosis. © 2003 Wiley‐Liss, Inc.
Is the soluble KDI domain of γ1 laminin a regeneration factor for the mammalian central nervous system?
Authors: Liebkind Ron , Laatikainen Timo , Liesi Päivi
Published: August 2003

Regeneration of adult mammalian CNS is poor as a result of environmental factors that prevent axon growth. The major factors hampering regeneration of central axons include proteins released from the damaged myelin sheets of the injured neuronal pathways and formation of the glial scar. By using an experimental model of human CNS injury, we show that survival and neurite outgrowth of human central neurons are significantly enhanced by the soluble KDI domain of γ1 laminin. Our results indicate that the KDI domain appears to neutralize both glia‐derived inhibitory signals and inhibitory molecules released from the myelin of the adult human spinal cord. We propose that the KDI domain may enhance regeneration of injuries in the adult mammalian CNS. © 2003 Wiley‐Liss, Inc.
Brain‐derived neurotrophic factor mitigates chronic ethanol‐induced attenuation of γ‐aminobutyric acid responses in cultured cerebellar granule cells
Authors: Ericson Mia , Haythornthwaite Alison R. , Yeh Pamela W.L. , Yeh Hermes H.
Published: August 2003

This study examined the effect of chronic exposure to ethanol and brain‐derived neurotrophic factor (BDNF) on the responsiveness of cerebellar granule cells to γ‐aminobutyric acid (GABA). Cerebellar granule cell cultures were chronically exposed to ethanol (100 mM), BDNF (20 ng/ml), or the combination of ethanol and BDNF. Whole‐cell current responses of granule cells to exogenously applied GABA were monitored following at least 5 days of chronic exposure. In the ethanol‐treated cultures, granule cell responsiveness to GABA was attenuated. Concomitant exposure of cultures to ethanol and BDNF mitigated the ethanol‐induced attenuation of GABA response, although BDNF, by itself, did not affect responsiveness to GABA. BDNF increased the expression of the GABAA receptor α6 subunit, whereas ethanol had no effect, in chronically treated granule cell cultures. In addition, concomitant treatment with BDNF and ethanol did not increase the expression of the GABAA receptor α6 subunit, so the subunit expression alone could not account for the mitigating effect of BDNF. We propose that different mechanisms regulating responsiveness to GABA underlie the effects induced by ethanol and BDNF, with the former influencing the expression of functional GABAA receptors and the latter involving the activation of the TrkB receptor and its downstream signaling pathways. © 2003 Wiley‐Liss, Inc.
Activation of the mitogen‐activated protein kinase pathway through p75NTR: A common mechanism for the neurotrophin family
Authors: Lad Shivanand P. , Neet Kenneth E.
Published: August 2003

Neurotrophins interact with two distinct classes of cell‐surface receptors, the Trk receptor tyrosine kinase family and the common neurotrophin receptor p75NTR. For many years, the biological role of p75NTR remained obscure, being relegated to modulating Trk binding of neurotrophins. Recently, the importance of p75NTR as a signaling receptor in itself has become increasingly clear. The signals initiated by p75NTR are likely to be as complex as those for the Trk family and probably depend on the cell system in which such signaling is being studied. In this study, all members of the neurotrophin family were demonstrated to be capable of stimulating p75NTR‐mediated activation of the mitogen‐activated protein kinase (MAPK) family (ERK1,2). This activation is rapid and transient, peaking at 5–15 min, depending on the cell system. The classical MAPK cascade consists of the reaction series Ras‐Raf‐MEK‐MAPK. The p75NTR‐induced MAPK activation is MEK dependent but Raf independent. This result implies that neurotrophin activation of p75NTR results in some cascade (as yet unknown) that bypasses Raf and converges on MEK to result in activation of MAPK. This activated MAPK is then able to translocate to the nucleus. The effect of this MAPK activation on cell survival is dependent on cell type. These results support the concept that signaling from the p75NTR receptor is more diverse and extensive than previously believed. © 2003 Wiley‐Liss, Inc.
Complementary DNA cloning and characterization of RANDAM‐2, a type I membrane molecule specifically expressed on glutamatergic neuronal cells in the mouse cerebrum
Authors: Kotani Masaharu , Tajima Youichi , Osanai Taka , Irie Atsushi , Iwatsuki Ken , Kanai‐Azuma Masami , Imada Masato , Kato Hiroko , Shitara Hiroshi , Kubo Hideo , Sakuraba Hitoshi
Published: August 2003

A membrane‐surface glycoprotein, RANDAM‐2, is one of the neuronal cell lineage‐specific antigens involved in the neuronal differentiation of P19 embryonic carcinoma (EC) cells and the mouse central nervous system (CNS). Complementary DNA cloning of RANDAM‐2 indicated that its nucleotide sequence completely matched that of PA2.26 antigen, a sialomucin‐like transmembrane glycoprotein previously found on tumorigenic keratinocytes. RANDAM‐2 transcripts were detectable from the embryonic stage of 6.5 days, and then the expression continued throughout the remaining embryonic stages and adulthood, with a localization restricted to the CNS. In growth factor‐induced neurospheres and adult cerebrum, RANDAM‐2‐expressing cells coincided well not only with nestin‐positive cells but also with glutamate‐positive neurons, but not with γ‐aminobutyric acid‐positive ones. These results indicate that RANDAM‐2 is one of the type I membrane surface antigens constitutively expressed on undifferentiated neuronal cells and the glutamatergic neuronal cells during mouse neurogenesis. © 2003 Wiley‐Liss, Inc.
Exogenous glutamate and taurine exert differential actions on light‐induced release of two endogenous amino acids in isolated rat retina
Authors: Barabás Péter , Kovács Ilona , Kardos Julianna , Schousboe Arne
Published: August 2003

A dark‐adapted isolated rat retina, preloaded with [14C]glutamate ([14C]Glu) and [3H]taurine ([3H]Tau), was superfused with artificial cerebrospinal fluid (ACSF) in the absence and presence of Glu (1 mM) or Tau (1 mM), as well as the Glu uptake inhibitors dihydrokainic acid (DHK, 0.04 mM) and trans‐L‐pyrrolidine‐2,4‐dicarboxylate (t‐PDC, 0.004 mM). After 10 min of light stimulation, the extracellular level of [14C]Glu and [3H]Tau was reduced to 82 ± 2% and 65 ± 4% of the control, respectively. Basal release was enhanced when Tau and t‐PDC were applied together, although none of the compounds had any effect when applied individually. Glu and DHK had no effect. The decrease of [14C]Glu efflux evoked by light stimuli was enhanced by t‐PDC and Tau, either added separately or together, whereas Glu and DHK were without effect. In contrast, [3H]Tau efflux variations induced by light stimuli were reduced markedly by both Tau and Glu. These findings suggest distinctly different roles of Tau and Glu in light‐induced responses in mammalian retina, including a possible role for Tau in light adaptation processes. © 2003 Wiley‐Liss, Inc.
Matrix metalloproteinase‐dependent shedding of syndecan‐3, a transmembrane heparan sulfate proteoglycan, in Schwann cells
Authors: Asundi Vinod K. , Erdman Robert , Stahl Richard C. , Carey David J.
Published: August 2003

Schwann cells transiently express the transmembrane heparan sulfate proteoglycan syndecan‐3 during the late embryonic and early postnatal periods of peripheral nerve development. Neonatal rat Schwann cells released soluble syndecan‐3 into the culture medium by a process that was blocked by inhibition of endogenous matrix metalloproteinase activity. When Schwann cells were plated on a substratum that binds syndecan‐3, the released proteoglycan bound to the substratum adjacent to the cell border. Membrane‐anchored syndecan‐3 was concentrated in actin‐containing filopodia that projected from the lateral edges of the Schwann cell membrane. Membrane shedding was specific for syndecan‐3 and was not observed for the related proteoglycan syndecan‐1. Analysis of Schwann cells transfected with wild‐type and chimeric syndecan‐1 and syndecan‐3 cDNAs revealed that membrane shedding was a property of the syndecan‐3 ectodomain. Inhibition of syndecan‐3 release significantly enhanced Schwann cell adhesion and process extension on dishes coated with the non‐collagenous N‐terminal domain of α4(V) collagen, which binds syndecan‐3 and mediates heparan sulfate‐dependent Schwann cell adhesion. Matrix metalloproteinase‐dependent syndecan‐3 shedding was also observed in newborn rat peripheral nerve tissue. Syndecan‐3 shedding in peripheral nerve tissue was age specific, and was not observed during later stages of postnatal nerve development. These results demonstrate that Schwann cell syndecan‐3 is subject to matrix metalloproteinase‐dependent membrane processing, which modulates the biological function of this proteoglycan. © 2003 Wiley‐Liss, Inc.
Macrophages contribute to the maintenance of stable regenerating neurites following peripheral nerve injury
Authors: Luk Hoenie W. , Noble Linda J. , Werb Zena
Published: August 2003

Normal adult uninjured nerve is unable to support axonal regeneration. We have studied the mechanisms underlying the regeneration of peripheral nerve by culturing adult mouse dorsal root ganglia (DRG) explants on unfixed, longitudinal cryosections of either the uninjured sciatic nerve or the distal segment of the transected sciatic nerve. We found that, initially, DRG grew vigorously on cryosections of both uninjured and postinjury sciatic nerves. However, the neurites began to degenerate shortly after contact with the uninjured nerve, whereas those growing on postinjury nerve substrate remained healthy for up to 9 days in culture. This ability to support stable outgrowth peaked at 8 days, gradually decreased by 10 days, and disappeared by 20 days after injury. Macrophages appeared in the distal segment by 4 days postinjury and had infiltrated its entire length by 8 days. Uninjured nerve cryosections could be rendered supportive of stable outgrowth by preincubation with macrophage‐conditioned medium or by brief trypsinization. The activity of the macrophage‐conditioned medium was augmented upon activation of macrophages. Together these findings suggest that the environment of the sciatic nerve undergoes a transformation during Wallerian degeneration such that it becomes transiently supportive of the stable outgrowth of neurites. This transformation may be mediated by a proteolytic activity, generated by activated macrophages, that removes a putative “degeneration signal” protein normally present in the adult nerve and thus contributes to the maintenance of stable regenerating neurites. © 2003 Wiley‐Liss, Inc.