bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of formylglycine in sulfatases by matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry
Authors: Peng Jianhe , Schmidt Bernhard , von Figura Kurt , Dierks Thomas 
Published: January 2003

AbstractCα‐Formylglycine, the catalytic amino acid residue in the active site of sulfatases, is generated by post‐translational modification of a cysteine or serine residue. We describe a highly sensitive procedure for the detection of Cα‐formylglycine‐containing peptides in tryptic digests of sulfatase proteins. The protocol is based on the formation of hydrazone derivatives of Cα‐formylglycine‐containing peptides when using dinitrophenylhydrazine as a matrix for matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOFMS). The hydrazone derivatives desorb and ionize with high efficiency and can be detected in the sub‐femtomole range. The presence of Cα‐formylglycine is indicated by a mass increment of 180.13 u, corresponding to the hydrazone moiety, and also by a unique C‐terminal fragment ion, characteristic of sulfatases, that becomes prominent in MALDI post‐source decay mass spectra of the hydrazone derivatives. Copyright © 2003 John Wiley & Sons, Ltd.
Effect of instrument tuning on the detectabilityof biopolymers in electrospray ionization mass spectrometry
Authors: Oberacher Herbert , Walcher Wolfgang , Huber Christian G. 
Published: January 2003

AbstractElectrospray ionization mass spectrometry of multiply charged biopolymer ions of different molecular size revealed a strong influence of tuning parameters on their detectability in quadrupole ion trap and triple quadrupole mass spectrometers. Hence, after optimizing the ion optical parameters with the signal of the 4− charge state of (dT)24 (low charge state tuning), a tenfold increase in the signal‐to‐noise ratio for a mixture of oligodeoxythymidylic acids (n = 12–18) was obtained compared with the results achieved with tune parameters optimized with a synthetic 80‐mer oligodeoxynucleotide. By contrast, a detection limit in the upper femtomole region could only be reached for a 104‐mer oligodeoxynucleotide utilizing the 24− charge state of the 80‐mer (high charge state tuning). The same effect was observed for proteins investigated in the positive ion mode using low and high charge states of cytochrome c and carbonic anhydrase, respectively, for instrument tuning. By comparing the settings for low and high charge state tuning, it became obvious that the most significant difference was observed in the potential applied to the heated metal capillary used to transfer ions from the atmospheric pressure to the vacuum region of the ion source. Taking advantage of the optimized tuning procedure, the molecular mass of a 61 base pair product of polymerase chain reaction was accurately determined by electrospray ionization mass spectrometry on‐line interfaced to ion‐pair reversed‐phase high‐performance liquid chromatography. Copyright © 2003 John Wiley & Sons, Ltd.
STAT1 and STAT3 α/β splice form activation predicts host responses in mouse hepatitis virus type 3 infection
Authors: Ning Qin , Berger Lloyd , Luo Xiaoping , Yan Weiming , Gong Feili , Dennis James , Levy Gary 
Published: January 2003

AbstractSignal transducer and activator of transcription 1α (STAT1 α) is reported to be essential for IFN‐γ and IFN‐α regulated gene expression, while STAT1 β, an alternate splice‐form, mediates only IFN‐α‐dependent gene expression. STAT3 α and STAT3 β splice forms are also differentially activated in response to cytokines including IL‐6 and IL‐10. The aim of this study was to investigate whether the STAT activation will predict the host immune response to viral infection and possibly a therapeutic target for the treatment of viral infection. Mouse hepatitis virus type 3 (MHV‐3) resistant strain (A/J) and sensitive mouse strains (BalB/cJ) were infected intraperitoneally (i.p.) with 100 plaque form units (pfu) of MHV‐3. The mice were sacrificed at the indicated times, and livers and spleens were immediately frozen in liquid nitrogen. Nuclear extracts proteins were detected by immunoblotting. STAT1 and STAT3 activation in spleen increased 24 to 72 hr following MHV‐3 infections in both sensitive and resistant mouse strains. However, over this time period, the ratio of activated α to β splice‐form for STAT1 and STAT3 increased above 1.0 in resistant A/J mice, while the ratio fell to <0.3 in MHV‐3 sensitive Balb/cJ and C3H/HeJ strains. Activated STAT1 α/β and STAT3 α/β ratio in liver were similar in resistant and sensitive mouse strains. Treatment of sensitive Balb/cJ mice with neutralizing anti‐TGF‐β antibody could increase the STAT1 α/β ratio to <1.0 in spleens, predicting enhanced rates of survival. These results suggested that ratio of activated STAT1 α/β and STAT3 α/β in mixed leukocytes from spleen predict the outcome to MHV‐3 infection, and may be an important marker and therapeutic target for modification of host immune response to virus infection. J. Med. Virol. 69:306–312, 2003. © 2003 Wiley‐Liss, Inc.
Kinetics of HBV DNA and HBsAg in acute hepatitis B patients with and without coinfection by other hepatitis viruses
Authors: Chulanov Vladimir P. , Shipulin German A. , Schaefer Stephan , Gerlich Wolfram H. 
Published: January 2003

AbstractThe kinetics of hepatitis B virus (HBV) and its surface antigen (HBsAg) during acute hepatitis has not yet been studied accurately in a representative number of patients. The influence of coinfecting hepatitis viruses during the acute phase of infection is not known. Three to four serum samples from 21 patients with acute HBV monoinfection and 27 with coinfection were taken at intervals of 6–10 days and analyzed for the number of HBV genome equivalents (ge) by real time polymerase chain reaction (PCR) and for HBsAg quantity using Laurell electrophoresis. Log HBV ge/ml decreased during the follow‐up from 6.8 ± 1.1 to 5.1 ± 1.0 to 4.2 ± 0.8 to 3.3 ± 1.1 (mean ± SD). The half‐life times of HBV ge increased from 1.6 days at the beginning to 4 days at the end. HBsAg decreased much slower: from 38 to 23 to 12 to3.8 μg/ml. Half‐life time was around 8 days at the beginning and 5.7 days at the end, but 11 patients showed a rapid elimination of HBsAg and HBV DNA. Hepatitis C virus (HCV) coinfection did not change the kinetics of HBV ge and HBsAg significantly. A moderate but significant suppression of HBV ge levels was observed in hepatitis D virus (HDV) coinfected patients. HBsAg levels were, however, enhanced in this cohort. In conclusion, the data suggest that expression and elimination of HBV is in most patients with acute hepatitis B not altered by coinfecting hepatitis viruses. The initial decrease of HBV ge and HBsAg in serum appears to be caused by decay or non‐specific removal in the absence of replacement. J. Med. Virol. 69:313–323, 2003. © 2003 Wiley‐Liss, Inc.
An outbreak of HBV and HCV infection in a paediatric oncology ward: Epidemiological investigations and prevention of further spread
Authors: Dumpis Uga , Kovalova Žanna , Jansons Juris , Čupane Liene , Sominskaya Irina , Michailova Marija , Karayiannis Peter , Gardovska Dace , Viazov Sergey , Ross Stefan , Roggendorf Michael , Pumpens Paul 
Published: January 2003

AbstractHospital‐acquired hepatitis B (HBV) and C virus (HCV) infections continue to occur despite increased awareness of this problem among the medical community. One hundred six patients were infected in a haematology oncology ward for children, over the time period 1996 to 2000. Serum samples from 45 such patients and 3 from infected medical personnel were used for nucleic acid amplification. HBV core, as well as HCV core and hypervariable region 1 (HVR1) nucleotide sequences, were analysed by phylogenetic tree analysis, in order to characterise the epidemiological pattern of viral transmission on the ward. Samples from 32 patients were positive for HBV‐DNA or HCV‐RNA by PCR. Ten patients were positive for both markers. Seventeen out of twenty‐three HCV core gene sequences were found to be evolutionarily related and clustered separately from other local sequences in the phylogenetic tree, indicating nosocomial transmission. This was confirmed by analysis of HVR1 gene sequences. One nurse and one physician from the ward were HCV RNA positive, but their HCV sequences were not related evolutionarily to those of the patient cluster. Fifteen out of nineteen HBV core gene sequences were also clustered together and were positioned separately in the relevant tree. Epidemiological investigation excluded a common source infection and indicated that spread of infection was most likely due to inappropriate infection control measures on the ward. No obvious risk factors for transmission were identified during the retrospective survey in patients with related sequences, except use of multidose vials for saline and poor staff compliance with routine hand hygiene procedures. The preventive measures that were introduced reduced the incidence of infection significantly. No new cases of HBV infection and only three anti‐HCV seroconversions occurred over a period of 19 months. The introduction and maintenance of strict prevention measures over a 2 year period, combined with HBV vaccination, reduced significantly the incidence of new HCV and HBV infections. J. Med. Virol. 69:331–338, 2003. © 2003 Wiley‐Liss, Inc.
Prevalence and clinical significance of circulating cryoglobulins in HIV‐positive patients with and without Co‐infection with hepatitis C virus
Authors: Fabris Paolo , Tositti Giulia , Giordani Maria Teresa , Romanò Luisa , Betterle Corrado , Pignattari Elena , Tagliaferri Cinzia , Muratori Paolo , Manfrin Vinicio , de Lalla Fausto 
Published: January 2003

AbstractAlthough hepatitis C virus (HCV) is a recognized cause of circulating cryoglobulins, the role of human immunodeficiency virus (HIV) in the pathogenesis of cryoglobulinemia has not been investigated extensively. To evaluate the prevalence of circulating cryoglobulins and to assess the relationship with clinical and virological parameters, 162 HIV‐positive subjects (84 anti‐HCV+) were tested for cryoglobulins, C3, C4, RF, autoantibodies, HIV‐viral titer, and CD4+ count. Anti‐HCV‐positive subjects were tested for HCV‐RNA, HCV‐viral titer, and HCV genotype. All patients were examined for the presence of signs or symptoms of vasculitis and tested for cryoglobulins using a standard biochemical assay. Cryoglobulins were found in 30 (18.5%) cases. Of the 30 positive cases, 29 (96.7%) were anti‐HCV‐positive and 28 (93.3%) HCV‐RNA‐positive. The presence of cryoglobulins was significantly associated (P < 0.01) with HCV‐RNA positivity (OR = 27), liver cirrhosis (OR = 16), decreased levels of C3 (OR = 8.6), C4 (OR = 13.6), increased levels of IgG and IgM (OR = 6.1 and 7.9, respectively), and RF positivity (OR = 6.3), but was unrelated to CD4+ cell count, HIV viral load, diagnosis of AIDS, HCV viral load and the presence of autoantibodies. Interestingly, the presence of cryoglobulins was not significantly associated with signs and symptoms commonly associated with cryoglobulinemia. In conclusion, HIV infection does not seem to play a significant role in the production of circulating cryoglobulins, which strongly correlates with HCV co‐infection and liver cirrhosis. Typical signs and symptoms of cryoglobulinemia do not correlate with the detection of circulating cryoglobulins in HIV and HCV patients. J. Med. Virol. 69:339–343, 2003. © 2003 Wiley‐Liss, Inc.
Intermittent detection of hepatitis C virus (HCV) in semen from men with human immunodeficiency virus type 1 (HIV‐1) and HCV
Authors: Pasquier Christophe , Bujan Louis , Daudin Myriam , Righi Laurence , Berges Laetitia , Thauvin Laurence , Berrebi Alain , Massip Patrice , Puel Jacqueline , Izopet Jacques 
Published: January 2003

AbstractHCV is usually transmitted via the blood, but HCV RNA has been detected recently in seminal fluid. This study was done to study HCV seminal shedding and factors that could influence the presence of HCV in the seminal fluid of men coinfected with HCV and HIV‐1. HCV and HIV‐1 genomes were assayed in multiple paired blood and semen samples obtained from 35 men enrolled in an assisted medical procreation protocol. HCV RNA was found intermittently in semen samples from 9 patients (25.7%). Samples from 9 men with HCV RNA in their semen and 26 men without were compared to further analyze these parameters. No correlation was found between HCV RNA in the seminal fluid and age, HCV virus load, the duration of HIV‐1 infection, HIV treatment, the CD4+ cell count, HIV‐1 virus load or HIV‐1 detection in the semen. The intermittent detection of HCV RNA in semen samples support the systematic search for HCV RNA in semen and the use of processed spermatozoa in assisted medical procreation of infertile HCV serodiscordant couples. J. Med. Virol. 69:344–349, 2003. © 2003 Wiley‐Liss, Inc.
Hepatitis C virus molecular epidemiology in Uzbekistan
Authors: Kurbanov Fuat , Tanaka Yasuhito , Sugauchi Fuminaka , Kato Hideaki , Ruzibakiev Ruslan , Zalyalieva Mariam , Yunusova Zulhumor , Mizokami Masashi 
Published: January 2003

AbstractThe aim of this study was to identify hepatitis C virus (HCV) genotypes and to estimate their prevalence in various risk groups and the regional distribution in Uzbekistan. Preliminary serological screening of 1,269 subjects revealed 6.5% anti‐HCV‐positive in a general population, 27.1% in patient groups, and 51.7% among intravenous drug users. HCV genotypes of 104 anti‐HCV‐positive subjects were determined using a PCR‐genotyping system in core region, and the results were supported by nucleotide sequencing of the NS5B region. Genotype 1b identified in total 64.2%, was the most prevalent. The genotype 3a identified in 25.0% was the second one distributed. HCV genotypes 2a, 1a, 2b, and 3b were identified in 3.8%, 2.9%, 2.9%, and 1.0% of cases, respectively. The intravenous drug users were distinguished from other groups by having the highest prevalence of genotype 3a, i.e., 50.0%, higher than the 33.3% for genotype 1b in this group. Geographically, genotype 1b was common; genotype 3a was also found frequently in all three regions. Uncommon HCV genotypes (1a, 2a, 2b, and 3b) were found in comparatively greater variability in the western region. Molecular evolutionary analysis based on the NS5B region did not reveal specific clustering or indigenous strains among Uzbekistan HCV isolates. In summary, two main mechanisms of HCV infection distribution were observed in Uzbekistan: HCV 1b genotype infection is widespread through blood products, and HCV 3a genotype infection is spreading through the growing number of intravenous drug users. J. Med. Virol. 69:367–375, 2003. © 2003 Wiley‐Liss, Inc.
Association of amino acid substitution pattern in nonstructural protein 5A of hepatitis C virus genotype2a low viral load and response to interferon monotherapy
Authors: Akuta Norio , Suzuki Fumitaka , Tsubota Akihito , Suzuki Yoshiyuki , Hosaka Tetsuya , Someya Takashi , Kobayashi Masahiro , Saitoh Satoshi , Arase Yasuji , Ikeda Kenji , Kumada Hiromitsu 
Published: January 2003

AbstractPatients with low titer (<0.5 mEq/ml) of hepatitis C virus (HCV) genotype 2a achieve high and sustained response (SR) rates to interferon (IFN) monotherapy, but we also encounter patients who are resistant to therapy. We explored the relationship between response to IFN and virological differences in such patients. We evaluated 159 consecutive naive patients with low titer of HCV genotype 2a who received IFN monotherapy. A case‐control study matched for age, sex, and viral load was conducted to examine the substitution patterns in amino acid positions (amino acids) 2163–2254 of nonstructural (NS) 5A between nonresponders to ideal IFN dose (≥500 million units) (nonresponders; NR) and responder to less than ideal dose. Overall, 82.4% achieved SR. The substitution numbers in amino acids 2193–2254 were higher in SR than NR patients (P < 0.05). High proportions of patients with substitution at amino acid 2205 (mainly threonine [T] instead of alanine [A]), dual amino acids 2169 and 2205 (mainly A‐T instead of T‐A), and those without substitution at amino acids 2227 were NR (P < 0.05). Four of 7 NR patients achieved SR after receiving a second course of IFN. Their amino acids patterns at positions probably associated with sensitivity to IFN did not change at the start of initial and second therapies except for one patient, and they had lower viral load and were treated with higher IFN dose in the second course compared with the initial course. Our results suggest that substitution patterns in NS5A in patients with low titer of HCV genotype 2a may affect their response to IFN, but the response to therapy may be affected by mechanisms other than substitutions in this region. J. Med. Virol. 69:376–383, 2003. © 2003 Wiley‐Liss, Inc.
Hepatitis C virus infection among pregnant women in Yaounde, Cameroon: Prevalence, viremia, and genotypes
Authors: Njouom Richard , Pasquier Christophe , Ayouba Ahidjo , Sandres‐Sauné Karine , Mfoupouendoun Jermie , Mony Lobe Marcel , Tene Gilbert , Thonnon Jocelyn , Izopet Jacques , Nerrienet Eric 
Published: January 2003

AbstractCentral Africa is considered to be an area of high endemic hepatitis C infection. To determine the prevalence of anti‐HCV antibodies, HCV RNA, and the genotype distribution in Cameroon, 1,494 pregnant women attending antenatal care units in Yaounde, Cameroon were screened for HCV infection. Anti‐HCV antibodies were detected with a 3rd generation ELISA (Monolisa anti‐HCV plus version 2, BioRad, Richmond, CA). All anti‐HCV antibody‐positive sera were then tested with another 3rd generation ELISA (AxSYM® HCV version 3, Abbott Laboratories, Abbott Park, IL) and subsequently for HCV RNA (Amplicor HCV, Roche Diagnostics, Basel, Switzerland). Genotype was determined by phylogenetic analysis of the NS5b gene. Seventy‐three pregnant women were found to be anti‐HCV antibody positive by the first ELISA, but only 28 were anti‐HCV positive by both ELISA. The prevalence of anti‐HCV antibodies was thus 1.9% (28/1,494) (95% CI: 1.3–2.7%). 21/28 (75%) of the positive samples by both ELISA were HCV RNA positive. The 45 samples that were HCV antibody negative by the second ELISA were also HCV RNA negative. The HCV subtypes identified were 1a (24%), 2f (38%) and 4f (38%). In contrast to previous studies, anti‐HCV antibodies were rare among pregnant women in Cameroon. The percentage of HCV seropositive pregnant women who had circulating HCV RNA was similar to that observed in Europe. Several HCV genotypes were found in Cameroon. J. Med. Virol. 69:384–390, 2003. © 2003 Wiley‐Liss, Inc.
Swine HEV infection in south India and phylogenetic analysis (1985–1999)
Authors: Arankalle V.A. , Chobe L.P. , Walimbe A.M. , Yergolkar P.N. , Jacob G.P. 
Published: January 2003

AbstractHepatitis E is endemic in India. It was recently noted that although all the Indian human hepatitis E virus (HEV) isolates (1976–2001) were placed in genotype I, the swine HEV recovered from western India (2000) belonged to genotype IV. This was in contrast to reports from the United States and Taiwan wherein both human and swine HEV belonged to the same genotype, i.e., genotypes III and IV, respectively. In order to validate these findings further, we retrospectively examined serum samples collected from pigs from southern India. Sequential serum samples from 45 (1985–1987) and 12 (1999) pigs from Karnataka state, south India, were screened for the presence of HEV RNA (nested PCR) and IgG‐anti‐HEV (ELISA). PCR products (Open Reading Frame‐2 region) were sequenced and subjected to phylogenetic analysis. In this study, 42/45 (1985–1987) and 12/12 (1999) pigs showed seroconversion to IgG anti‐HEV antibodies, with a mean age at seroconversion of 4.8 ± 1.6 months. Four samples collected in 1999 and two samples collected during 1985 were HEV RNA positive. All swine HEV sequences clustered with genotype IV, demonstrating that swine HEV was prevalent among south Indian pigs for at least for 16 years and, similar to western India, belonged to genotype IV. Thus, genotype I and IV HEV continue to circulate in humans and pigs, respectively, from India. Whether swine HEV infects humans remains to be determined. J. Med. Virol. 69:391–396, 2003. © 2003 Wiley‐Liss, Inc.
Polymerase chain reaction as a rapid diagnostic tool for therapy of acute retinal necrosis syndrome
Authors: Gargiulo Franco , De Francesco Maria Antonia , Nascimbeni Giuseppe , Turano Raffaele , Perandin Francesca , Gandolfo Enrico , Manca Nino 
Published: January 2003

AbstractHerpesviruses are involved in the pathogenesis of acute retinal necrosis syndrome (ARN). A rapid and accurate diagnosis of herpetic infections is crucial for prompt administration of a specific antiviral therapy. The purpose of this study was to evaluate a polymerase chain reaction (PCR)‐based assay to detect herpesvirus DNA in the aqueous humor of clinical samples from ten patients with uveitis and clinical suspicion of ARN. Samples were assayed for herpes simplex virus type 1‐2 (HSV 1‐2), varicella zoster virus (VZV) and cytomegalovirus (CMV). Clinical suspicion of ARN was confirmed for four patients. Two patients (one with bilateral ARN) tested PCR‐positive for VZV DNA and the other two were positive for HSV 1‐2 DNA. CMV DNA was not detected in any of the samples, and no sample was positive for DNA from more than one virus. The remaining patients did not show any evidence of herpesvirus DNA in their aqueous samples. Our findings demonstrate that the use of PCR for detecting herpesvirus DNA in aqueous humor of uveitic subjects may be a valuable tool for early diagnosis of acute retinal necrosis syndrome and for timely administration of a suitable therapy. J. Med. Virol. 69:397–400, 2003. © 2003 Wiley‐Liss, Inc.
Genetic characterization of new Dobrava hantavirus isolate from Greece
Authors: Nemirov Kirill , Vapalahti Olli , Papa Anna , Plyusnina Angelina , Lundkvist Åke , Antoniadis Antonis , Vaheri Antti , Plyusnin Alexander 
Published: January 2003

AbstractThe first complete genome sequence of Dobrava hantavirus isolated from yellow‐necked mouse Apodemus flavicollis trapped in the northeastern Greece is described. The S, M, and L segments of the Greek isolate of Dobrava virus are 1673, 3635, and 6532 nucleotides (nt) long, respectively, and encode the nucleocapsid (N) protein of 429 amino acids (aa), glycoprotein precursor of 1135 aa, and the L protein of 2151 aa. N protein contains three cysteine residues conserved in all known hantaviruses, as well as structural domains responsible for the RNA binding and presumable interaction with the apoptosis enhancer Daxx. All cysteine residues and glycosylation sites that are conserved among G1G2 sequences of all hantaviruses species were also found in the Greek isolate. The L protein contains all the polymerase motifs and structural domains found in other hantavirus polymerases. Comparison of the Greek isolate of Dobrava virus with other hantaviruses showed the highest level of sequence homology with Dobrava virus isolate from Slovenia. Other hantaviruses carried by Murinae rodents (Saaremaa, Hantaan, Seoul, and Thailand viruses) were more divergent and hantaviruses carried by Arvicolinae or Sigmodontinae rodents showed the highest genetic diversity with the Greek isolate of Dobrava. The results of phylogenetic analyses confirmed these observations and showed a monophily of all the Dobrava virus strains that, in turn, shared more ancient ancestors first with Saaremaa virus and then with other Murinae‐borne hantaviruses. J. Med. Virol. 69:408–416, 2003. © 2003 Wiley‐Liss, Inc.