bluuebunny/crossref_metadata_2025 huggingface dataset:
Genetic diversity of hepatitis B virus strains isolated in Rio de Janeiro, Brazil
Authors: Niel C. , Moraes M. T. B. , Gaspar A. M. C. , Yoshida C. F. T. , Gomes S. A.
Published: March 2007

Hepatitis B virus (HBV) genomes have been classified into six groups, A–F, group F being the most divergent. South America has provided the smallest number of strains studied at the genome level. The only HBV strain from this region sequenced completely has been classified in group F, and contains the most divergent of the HBV genomes presently known [Naumann et al. (1983): Journal of General Virology 74:1627–1632]. To evaluate genetic relatedness between strains isolated in South America and in the rest of the world, a restriction endonuclease analysis was carried out on 14 HBV strains (4 adw2, 3 adw4, 3 ayw2, and 4 ayw3) isolated in Rio de Janeiro, Brazil. C, pre‐S, and X genes along with the 5′ part of the P gene from these strains were amplified by the polymerase chain reaction. The DNA fragments were digested by BamHI, BstEII, EcoRI, Hhal, and Taql endonucleases. The restriction patterns obtained were compared with those deduced from the nucleotide sequence of 26 HBV strains isolated in other continents. The results showed a large genetic variability of Brazilian strains. Taking into account that all the samples examined possessed the w antigenic subdeterminant, the number of different restriction patterns of strains isolated in Rio de Janeiro was at least as large as that of the reference strains isolated in the rest of the world. Some original restriction patterns were found in adw4 and ayw2 HBV strains.
Hepatitis may precede the occurrence of precore region mutation in hepatitis B virus genome during infection in young carriers
Authors: Xuan‐Thanh Nguyen , Fukuda Ryo , Fukumoto Shiro
Published: March 2007

To understand when the mutation with a stop codon of precore region in hepatitis B virus genome occurs, the prevalence of the mutation of viral DNA clones propagated from sera of school‐age carriers was investigated with respect to hepatitis B e antigen (HBeAg)/anti‐HBe and sequential changes of mutants along HBeAg seroconversion were analyzed.

Of 32 carriers aged 8–18 years, 14 HBeAg(+) patients had 2.2% mutant clones, whereas 8 patients with low titer anti‐HBe had a higher rate of 18.1% (P< 0.01) and the highest rate of 61.3% was found in 10 patients with high anti‐HBe titer (P< 0.001). By contrast, the amount of viral DNA decresed significantly in patients with anti‐HBe. Sequential analysis in six cases revealed three types of seroconversion with time difference of the emergence and increase of mutant clones.

It is concluded that mutation occurs at a relatively young age and increases along time and/or HBeAg seroconversion. Hepatitis might precede or accelerate the emergence and increse of mutant population which might be predictive of sustained resolution of the disease.
Detection of varicella‐zoster virus DNA in blood of children with varicella
Authors: Ozaki Takao , Kajita Yuji , Asano Yoshizo , Aono Toshiya , Yamanishi Koichi
Published: March 2007

The nested polymerase chain reaction was used to detect varicella zoster virus (VZV) DNA in 67 samples of peripheral blood mononuclear cells (PBMC) from 39 otherwise healthy children with varicella. Eleven were during the incubation period and 56 were after appearance of rash. VZV DNA was detected in two of eight (25%) PBMC at the early phase of the incubation period (day –14 to –11). The rate of the detection increased to 67% in –5 to –1 days prior to the onset of the rash and 46% in 0–4 days after the onset. It declined gradually with time and was undetectable in patients after 15 days from the clinical onset. The serum antibody determined with the fluorescent antibody to membrane antigen was first detected in the samples on day 2 and subsequently increased to 100% rapidly. In conclusion, VZV DNA cannot be detected usually in PBMC of healthy children except in varicella. © 1994 Wiley‐Liss, Inc.
Genomic diversity of small round structured viruses in the United Kingdom
Authors: Norcott J. P. , Green J. , Lewis D. , Estes M. K. , Barlow K. L. , Brown D. W. G.
Published: March 2007

Fifty‐two fecal specimens collected in the United Kingdom between 1986 and 1992, which contained small round structured virus (SRSV) particles, were tested by reverse transcriptase polymerase chain reaction assays using two primer pairs derived from sequences of Snow Mountain Agent and Norwalk virus. There was poor correlation between results obtained with each primer pair. Twenty specimens (38%) gave positive bands with SM51/31 primers and 18 (34%) were positive with SM52/32 primers, with a total of 30 specimens (57.7%) giving amplification products of the expected size with one or both primer pairs. Genomic variation was investigated by sequencing a 266 bp region of the RNA polymerase gene from nine strains which had been antigenically typed by solid phase immune electron microscopy (SPIEM). RNA sequence identities ranged from 53 to 99%. Three genomic groups were suggested by phylogenic analysis, the first of which contained Norwalk virus, Southampton virus, and strains typed by SPIEM as SRSV UK2. The second contained Snow Mountain agent and strains typed as either SRSV UK3 or UK4. The third contained strains typed as SRSV UK1 and strains untypeable by SPIEM. Some correlation was demonstrated when antigen typing by SPIEM and phylogenic grouping based on sequence data were compared. © 1994 Wiley‐Liss, Inc.
Investigation of cytomegalovirus infection as a risk factor for coronary atherosclerosis in the explanted hearts of patients undergoing heart transplantation
Authors: Dummer Stephen , Lee Ann , Breinig Mary Kay , Kormos Robert , Ho Monto , Griffith Bartley
Published: March 2007

Infection with a human herpes virus, particularly cytomegalovirus (CMV), has been hypothesized to be a cofactor in the development of atherosclerosis in humans. We investigated the association of prior CMV infection with the presence of atherosclerosis in the coronary arteries of the native heart of 314 individuals undergoing heart transplantation. Age, male gender, race, tobacco use, and previous general and cardiac surgery were also studied as covariables. Factors associated with the presence of coronary atherosclerosis by univariate analysis were age greater than the median of 48 years (odds ratio [OR] = 5.9, 95% confidence intervals [Cl] 3.0–11.6; P < 0.0001), tobacco use (OR = 3.8, 95% Cl 2.1–7.0; P < 0.005), CMV seropositivity (OR = 3.1, 95% Cl 1.8–5.5; P < 0.001), and male gender (OR = 3.0, 95% Cl 1.6–5.4; P < 0.0005). When patients were divided into quartiles based on age, coronary atherosclerosis was shown to be associated with CMV seropositive status only in the youngest quartile of patients (OR = 3.6, 95% Cl 1.4–8.9; P < 0.01) but not in the older three quartiles of patients (OR = 0.9, 95% Cl = 0.3–2.4; P > 0.5). In multiple logistic regression analyses, CMV seropositivity was not a significant independent variable in the whole group of patients (P = 0.13) but remained a significant variable in the youngest quartile of patients (P = 0.01). However, 43% of these younger patients and 29% of all patients with coronary atherosclerosis were seronegative for CMV. We infer that prior CMV infection is not a general risk factor for atherosclerosis, but it may be a cofactor in a subset of younger patients. Further studies are warranted but need to be carefully controlled for confounding variables. © 1994 Wiley‐Liss, Inc.
Maturation of antibody avidity after primary human cytomegalovirus infection is delayed in immunosuppressed solid organ transplant patients
Authors: Lutz E. , Ward K. N. , Gray J. J.
Published: March 2007

An IgG antibody avidity assay which uses urea to modify a commercial enzyme‐linked immu‐nosorbent assay (ELISA) has been investigated for its ability to distinguish primary human cy‐tomegalovirus (CMV) from recurrent or long‐term infection. Twenty‐four immunosuppressed solid organ transplant patients were studied. The avidity indices for IgG to CMV were low for 12 out of 13 patients with primary infection (mean 18%), high for all 11 patients with long‐term infection (mean 85%), and the 1 patient with primary infection showing an intermediate avidity index (51%) was found to have acquired passively large amounts of CMV immunoglobulin, presumably of high avidity, during therapy.

From the results, low and high avidity indices were defined as lying between 0–34% and 60–100%, respectively, and it was thus clear that the avidity assay can discriminate between primary and recurrent or long‐term CMV infection. The avidity indices of eight of the immunosuppressed organ transplant patients with primary infection were followed in serial serum samples over time and IgG antibody to CMV was found to take at least a year to mature to high avidity in contrast to the 2–6 months expected for normal subjects. This finding provides evidence that im‐munosuppression has subtle, hitherto unsuspected, effects on humoral immunity to CMV in addition to the well‐known depression of cell‐mediated responses. It is concluded that this reliable avidity assay will be of importance in the diagnosis of CMV infection and in elucidating the pathogenesis of CMV‐induced disease in organ transplant recipients. © 1994 Wiiey‐Liss, inc.
E2 and NS5: New antigens for detection of hepatitis C virus antibodies
Authors: Zaaijer H. L. , Vallari D. S. , Cunningham M. , Lesniewski R. , Reesink H. W. , Van Poel C. L. Der , Lelie P. N.
Published: March 2007

The value of two new hepatitis C virus (HCV) antigens for detection of HCV antibodies was studied. These two recombinant antigens were derived from the nonstructural‐5 (NS5) and envelope ‐2 (E2) region of the HCV genome. In a panel of 33 HCV‐RNA positive samples with indeterminate Riba‐2 confirmatory test results, 29 samples (88%) showed additional antibody reactivity against E2 and 12 samples (36%) snowed additional reactivity against NS5. Among 39 HCV‐RNA negative, Riba‐2 indeterminate donor samples, no additional E2 or NS5 reactivity was found in 34 samples (87%); while 5 samples (13%) showed additional reactivity against NS5 and/or E2. E2 reactivity thus resolved the majority of hitherto indeterminate samples. In serial samples from nine posttransfusion hepatitis C patients, NS5 and E2 antibodies did not appear earlier than classical HCV antibodies. However, E2 antibodies eventually appeared in all nine patients. The recombinant E2 might be a candidate antigen for future HCV antibody assays. © 1994 Wiley‐Liss, Inc.
Detection of hepatitis C virus genome in formalin‐fixed paraffin‐embedded liver tissue by in situ reverse transcription polymerase chain reaction
Authors: Lau George K. K. , Fang Jane W. S. , Wu P. C. , Davis Gary L. , Lau Johnson Y. N.
Published: March 2007

Detection of hepatic hepatitis C virus RNA and antigens is difficult since their expression is very low. The technique of in situ reverse transcription polymerase chain reaction (IS‐RT‐PCR) was developed for the detection and localization of HCV RNA in formalin‐fixed paraffin‐embedded liver sections. Key steps for achieving a good signal‐to‐background ratio include appropriate proteolytic digestion, DNase pretreatment, higher Mg2+ concentration, primers from the core region, „hot‐start”︁, and between 10–20 cycles of PCR. Using this approach, HCV RNA was detected in the liver in four of the six patients. In those positive samples, the percentage of cells positive for HCV RNA ranges from <5 to 25%. Using RT‐PCR of serum samples and RNA extracted from the corresponding tissue block (10 sections) as the gold standard and sections from four control subjects as negative controls, the sensitivity and specificity of this assay was 4/6 and 4/4, respectively. The specific positive signal was found mainly in the cytoplasm of the hepatocytes and occasionally in mononuclear cells but not in the bile duct epithelium or sinusoidal cells. These positive hepatocytes were scattered in the liver lobules with occasional clustering. These preliminary data confirm the hepatotropic nature of HCV and the localization of HCV RNA is consistent with cytoplasmic replication. © 1994 Wiley‐Liss, Inc.
Single dose inactivated hepatitis A vaccine: Rationale and clinical assessment of the safety and immunogenicity
Authors: van Damme P. , Matheï C. , Thoelen S. , Meheus A. , Safary A. , André F. E.
Published: March 2007

In comparison with the classical immunisation schedules (0‐1‐6 or 0‐1‐12 months) for hepatitis A, a 0‐ and 12‐ or a 0‐ and 6‐month schedule would have important advantages by reducing the number of injections and discomfort and increasing scheduling convenience and patient compliance. It would be convenient if a single dose with enough antigen could protect both rapidly and for at least 12 months, when the booster dose would be given. Several clinical trials have been carried out with an inactivated hepatitis A vaccine containing 1,440 ELU. (1 ml), according to a 0–12 and a 0–6 vaccination schedule.

This hepatitis A vaccine is safe and well tolerated. It offers a rapid seroresponse: 14 days after a single dose the seroconversion is 88% (95% C.I.: 84.6–90.9). The 0–12 schedule study showed good persistence of hepatitis A virus (HAV) antibodies with a seroconversion rate of almost 95% at month 12. Booster doses given at 6 or 12 months result in a substantial rise in antibody levels; according to these antibody litres, the 1,440 EL. U. vaccine can be expected to confer comparable duration of protection as the 720 EL. U. vaccine, i.e., 10–20 years. Preliminary data show that tinning of the booster may not be critical for the antibody response.

In conclusion, the 1,440 EL. U. hepatitis A vaccine is safe, offers rapid seroconversion, and is highly immunogenic. The persistence of HAV antibodies until month 12 allows a certain flexibility in the administration of the booster: month 6 or 12, and a 0–12 or 0–6 schedule can increase the vaccination compliance. copy; 1994 Wiiey‐Liss, inc.
Effects of ribavirin on intrahepatic and extrahepatic expression of hepatitis C virus in interferon nonresponsive patients
Authors: Koskinas John , Tibbs Christopher , Saleh Mohammed G. , Pereira Leila M. M. B. , McFarlane Ian G. , Williams Roger
Published: March 2007

Response to ribavirin therapy (1,000‐1,200 mg/day for 6 months) was evaluated in nine patients with chronic hepatitis C virus (HCV) infections who had previously failed to respond to a 6‐month course of alpha‐interferon. All had chronic active hepatitis with elevated serum aminotransferase activities (mean ± SD = 138 ± 66IU/I). During ribavirin therapy, three showed a complete response (normalized serum ami‐notransferase), although in one patient this returned to the pretreatment level 2 months after treatment was stopped. Three others showed a partial response (serum aminotransferase reduction by ⩾50;%) and the remainder showed no response. There were no consistent changes in HCV‐RNA (positive strand) in serum, liver, or peripheral blood mononuclear cells during therapy, but two patients lost HCV‐RNA from serum and three of five patients with negative strand HCV‐RNA in their livers lost this putative replicative form of the virus. The findings suggest that ribavirin may exert its effects by suppressing viral replication rather than by eradicating the virus, at least in this group of patients, and that the drug may have some benefit in selected cases of chronic hepatitis C that are resistant to inter‐feron. However, peripheral blood mononuclear cells represent a major extrahepatic reservoir of HCV and the present regimen of ribavirin therapy did not significantly affect this situation. More prolonged therapy may be required to eradicate the virus from this large pool of cells with the potential to continually reinfect the liver. © 1995 Wiley‐Liss, Inc.
Lack of augmenting effect of interferon‐γ on dengue virus multiplication in human peripheral blood monocytes
Authors: Sittisombut Nopporn , Maneekarn Niwat , Viputtikul Kwanjai , Supawadee Jiraporn , Kanjanahaluethai Amornrat , Kasinrerk Watchara
Published: March 2007

The effect of interferon—γ (IFN‐γ) on dengue virus multiplication in human peripheral blood mono‐cytes was investigated. Enriched monocytes were treated with IFN‐γ and then infected with dengue virus type 2 either directly or in the presence of optimal infection‐enhancing levels of antibodies. Pretreatment of monocytes from dengue‐immune donors with 100 IU/ml of IFN‐γ caused 12‐ to 97‐fold and 13‐ to 137‐fold reduction of virus yields at 24 hr after infection in the absence and presence of an anti‐flavivirus monoclonal antibody, respectively. IFN‐γ also diminished virus yields when infection of monocytes from a donor who lacked anti‐dengue antibody was enhanced 40‐fold. The percentage of infected monocytes in IFN‐γ‐pretreated cultures was similarly reduced. Dominance of the antiviral effect of IFN‐γ in monocytes is in contrast to an augmenting effect previously observed in the promonocytic cell line U937. © 1995 Wiley‐Liss, Inc.
Evaluation of a multiple peptide assay for typing of antibodies to the hepatitis C virus: Relation to genomic typing by the polymerase chain reaction
Authors: Zhang Zhu‐Xu , Yun Zhi‐Bing , Chen Margaret , Sönnerborg Anders , Sällberg Matti
Published: March 2007

A panel of 16 type‐specific synthetic peptides corresponding to variable antigenic regions within the hepatitis C virus (HCV) core, nonstruc‐tural 4 (NS4), and NS5 proteins was synthesised. The peptide panel was used to develop an enzyme immunoassay (EIA) for the detection of antibodies directed to HCV type 1 (genotypes I/1a and II/1 b), type 2 (genotypes III/2a and IV/2b), and type 3 (genotype V/3). The peptides corresponded to residues 68–81 of the HCV core (types 1,2, and 3), residues 1692–1705 and 1710–1728 of HCV NS4 (types 1 a, 1 b, 2a, 2b, and 3), and residues 2303–2319 of HCV NS5 (types 1a, 1b, 2a, and 2b). The 16‐peptide panel was evaluated using human sera from 46 carriers of HCV, which were genotyped in parallel by the polymerase chain reaction (PCR) using primers specific for types I, II, III, IV, and V of HCV core. Of the 46 carriers, 14 (30%) were infected by HCV genotype I, 7 (15%) by genotype II, 16 (35%) by HCV genotype IV, and 6 (13%) by HCV of genotype V. Two carriers had double infections of types I and II, and the HCV strain of one carrier could not be genotyped. Using the serotyping system, 40 (89%) out of the 45 genotyped carriers were found to contain type‐specific antibodies corresponding to the genotypes identified by PCR. In 5 of the 23 carriers infected by genotypes I and/or II, antibodies specific for HCV type 1 could not be detected, whereas all 16 carriers infected by genotype IV were serologically typed as type 2. Out of the six carriers infected by HCV of genotype V, all were found to have antibodies of serotype 3, but in most cases together with antibodies to NS5 type 1, indicating sequence homologies between types 1 and 3 of this NS5 region. In the one patient serum where the HCV strain could not be genotyped, a mixture of types 1, 2, and 3 antibodies were found. In conclusion, a serotyping system with a sensitivity and specificity of 89% was developed. It is confirmed that at least three distinct serotypes of antibodies to HCV exist. The major advantage of using four different antigenic regions is that we often obtain high absor‐bance values which are easily interpreted, or multiple reactions which confirm each other. © 1995 Wiley‐Liss, Inc.
Epidemiology of Norwalk virus during an outbreak of acute gastroenteritis aboard a US aircraft carrier
Authors: Sharp Trueman W. , Hyams Kenneth C. , Watts Douglas , Savarino Stephen J. , Trofa Andrew F. , Martin Gregory J. , Kapikian Albert Z. , Green Kim Y. , Jiang Xi , Estes Mary K. , Waack Matthew
Published: March 2007

A large outbreak of acute gastroenteritis occurred over a 5‐week period aboard an aircraft carrier. The estimated cumulative attack rate was 13% among the 4,500‐man crew. Eight percent of the crew sought medical attention, nearly all of whom missed 1 day or more of work. The risk of developing illness was 2 to 3 times greater for individuals living in more crowded sleeping quarters ( >50 persons per compartment). Occurrence of gastroenteritis was associated with a fourfold or more rise in Norwalk virus antibody levels, as measured by an enzyme‐linked immunoassay utilizing a baculovirus expressed recombinant antigen. In addition, 27 nm Norwalk virus‐like particles were visualized in two of six stools examined by immune electron microscopy. The presence of a low (<1:50) or a high (⩽1:6,400) pre‐illness antibody level was associated with a lower incidence of illness. This investigation indicates that Norwalk virus can adversely impact operations of a military vessel and that crowding is a major risk factor in transmission. © 1995 Wiley‐Liss, Inc.
Differentiation of vaccine and wild mumps viruses by polymerase chain reaction and nucleotide sequencing of the SH gene: Brief report
Authors: Künkel Udo , Driesel Gerhard , Henning Ursula , Gerike Edith , Schreier Eckart , Willers Hildegard
Published: March 2007

The nucleotide sequences of the SH gene and its F gene flanking region were determined over a range of 322 nucleotides for two live vaccines, two vaccine‐associated isolates, six wild mumps viruses, and an attenuated mumps virus and compared to other sequences already published. Comparison revealed that the vaccine strains were clearly different from each other and the postvaccination isolates were different from the vaccines used. The viruses were assigned to three known cocirculating viral lineages. The attenuated mumps virus possesses nucleotide sequences identical to those of its progenitor Strain. © 1995 Wiley‐Liss, Inc.
Complete nucleotide sequences and the characteristics of two hepatitis B virus mutants causing serologically negative acute or chronic hepatitis B
Authors: Uchida Toshikazu , Gotoh Kenichiro , Shikata Toshio
Published: March 2007

Hepatitis B virus (HBV) DNA was amplified by the polymerase chain reaction from the sera of a patient with acute hepatitis and a patient with chronic hepatitis. Both patients were negative for serum hepatitis B surface antigen and hepatitis B core antibodies and had been previously diagnosed as non‐A, non‐B, non‐C, non‐D, non‐E hepatitis. The nucleotide sequence revealed an 8‐nucleotide deletion in the X‐gene coding region creating a C‐terminally truncated X protein, and probable mutation of the enhancer 11/core promoter element. In addition, DR2 showed a T‐to‐C mutation at the extreme 5′‐terminus. These mutations within the X‐gene coding region must suppress replication and expression of HBV DNA, and this seems to be responsible for absence of serological markers despite the presence of HBV infection. © 1995 Wiley‐Uss, inc.
Human T‐cell lymphotropic virus type I in Iranian‐born Mashhadi Jews: Genetic and phylogenetic evidence for common source of infection
Authors: Nerurkar V. R. , Song K. J. , Melland R. R. , Yanagihara R. , Achiron A. , Melamed E. , Shohat B. , Pinhas‐Hamiel O.
Published: March 2007

High prevalence of human T‐cell lymphotropic virus type I (HTLV‐I) infection and disease has been identified among Iranian‐born Mashhadi Jews, an ethnically segregated, highly inbred population. To determine the origin and genetic diversity of HTLV‐I in this group, 1,039 bp spanning selected regions of the HTLV‐I gag, pol, env and pX genes were enzymatically amplified and sequenced directly from DMA of five Mashhadi Jews (three with spastic myelopathy and two asymptomatic carriers). Alignment and comparison of these sequences with cosmopolitan and Australo‐Melanesian topotypes of HTLV‐I indicated that the HTLV‐I strains from Mashhadi Jews, which were ⩾99.9% identical among themselves, exhibited considerable sequence similarity (⩾99%) to HTLV‐I strains from southern India, suggesting a common source of infection. Phylogenetic analysis, using the maximum parsimony method, was consistent with a single‐source introduction of HTLV‐I into the Mashhadi Jewish community. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus infection in schistosomiasis mansoni in Brazil
Authors: Pereira Leila M. M. B. , Saleh Mohammed G. , Koskinas John , Williams Roger , McFarlane Ian G. , Melo Marcelo C. V. , Domingues Ana L. C. , Spinelli Victorino , Mies Sérgio , Massarolo Paulo
Published: March 2007

The involvement of the hepatitis C virus (HCV) in the severity of liver disease in chronic schistosomiasis was investigated in 215 Brazilian patients with S. mansoni infections, but without evidence of hepatitis B surface antigen (HBsAg). Forty‐three had hepatointestinal (HIS) and 172 had hepatosplenic schistosomiasis (HSS), and 135 had compensated (HSSC), and 37 had decompensated (HSSD) liver disease. Fifty‐two (24%) were found to have evidence of HCV infection (sero‐positive for anti‐HCV antibodies and/or HCV‐RNA). These comprised 35 (95%) of the 37 with HSSD, 16 (12%) of the 135 with HSSC, and 1 (2.4%) of the 43 with HIS, compared with only 1 (2%) of 50 control patients without S. mansoni. Testing of matched liver tissue and peripheral blood mononuclear cells (PBMCs) from 25 patients (6 HSSC and 19 HSSD) with HCV infections showed that 17 (68%) had “active” viral infections, in that negative strand HCV‐RNA (the presumed replicative intermediate of the virus) could be detected in liver and/or PBMCs. Among these 25, negative strand HCV‐RNA was found in 16 (84%) of the 19 with chronic active hepatitis, but in only 1 (17%) of the 6 with mild or inactive disease (P <0.01). HCV‐RNA was detected in matched spleen specimens from 9 of 10 patients (all of whom were also positive in PBMCs), suggesting that the spleen is an important extrahepatic reservoir of the virus. The findings indicate that concomitant infections with HCV are a major factor contributing to the severity of liver disease in chronic schistosomiasis, and that apparent active replication of the virus in the liver or at extrahepatic sites is strongly associated with severe liver disease. © 1995 Wiley‐Liss, Inc.
Efficacy of Kakkon‐to, a traditional herb medicine, in herpes simplex virus type 1 infection in mice
Authors: Nagasaka Kazuhiko , Kurokawa Masahiko , Imakita Masami , Terasawa Katutosi , Shiraki Kimiyasu
Published: March 2007

Kakkon‐to is one of the representative traditional herb medicines (Kampo formulae) and has been used historically for the treatment of infectious diseases in China and Japan. The efficacy of this preparation was characterised using a cutaneous herpes simplex virus type 1 (HSV‐1) infection in mice as a model for human viral infection. Kakkon‐to at a dose corresponding to human use reduced significantly the mortality of HSV‐1‐infected mice and localised skin lesions. Delayed type hypersensitivity (DTH) response to HSV‐1 antigen was significantly stronger in treated mice than in untreated mice. However, no histopathological difference was noted in the skin lesions between treated and untreated mice except for the size of the lesions. Kakkon‐to did not inhibit the growth of HSV‐1 in vitro. Natural killer cell activity, natural cytotoxic killer cell activity, and the population of T‐cell subsets in spleen cells of infected mice were not affected by the drug. Kakkon‐to did not augment interferon induction and anti‐HSV‐1 antibody production, nor increased cytokine levels such as interleukin‐1α, interleukin‐2, interferon‐γ, and tumour necrosis factor‐α in sera of infected mice. Thus, Kakkon‐to induced strong DTH to HSV‐1 in infected mice, which may have caused localisation of skin lesions and reduction in the mortality of treated mice. © 1995 Wiley‐Liss, Inc.
Immunogenicity and safety of an inactivated hepatitis a vaccine in anti‐HIV positive and negative homosexual men
Authors: Hess Georg , Clemens Ralf , Bienzle Ulrich , Schönfeld Christian , Schunck Birgit , Bock Hans‐Ludwig
Published: March 2007

The immunogenicity, reactogenicity, and safety of an inactivated hepatitis A vaccine were assessed in anti‐HIV positive homosexual men. Fourteen anti‐HIV positive (group 1) and 20 anti‐HIV negative (group 2) men received vaccine (containing 720 ELISA units of hepatitis A antigen per dose) intramuscularly at 0, 1, and 6 months. Twelve unvaccinated anti‐HIV positive men (group 3) were included as controls to evaluate disease progression.

Seroconversion (anti‐hepatitis V virus (HAV ⩾20 mlU/ml) was higher in group 2 than group 1 at months 2 (100% vs. 73%) and 7 (l00%vs. 77%). Group 2 had higher antibody titres than group 1 at months 1 (201 vs. 92 mlU/ml) and 7 (1, 687 vs. 636 mlU/ml). The decline in CD4+ cells between months 0 and 7 was similar in groups 1 and 3 (6.4% vs. 16.2%), showing no evidence for harmful effect of the vaccine on the course of HIV infection. This hepatitis A vaccine appears safe, well tolerated, but less immunogenic in HIV positive homosexual men. © 1995 Wiley‐Liss, Inc.
Measurement by the polymerase chain reaction of the Epstein‐Barr virus load in infectious mononucleosis and AIDS‐related non‐Hodgkin's lymphomas
Authors: Laroche Christine , Drouet Emmanuel B. , Brousset Pierre , Pain Christiane , Boibieux Andre , Biron Francois , Icart Josette , Denoyel Gerard A. , Niveleau Alain
Published: March 2007

A polymerase chain reaction (PCR) assay for the detection of Epstein‐Barr virus (EBV) sequences in various clinical samples, especially peripheral blood leukocytes (PBL) and serum, was carried out and the results obtained were compared with specific EBV serology. One hundred seventy patients were enrolled in the study: 89 healthy blood donors, 22 asymptomatic patients, 36 individuals with primary EBV infection (including 19 patients with infectious mononucleosis [IM]), 22 HIV‐infected subjects (including 4 with hairy oral leukoplakia, 3 with central nervous disorders, and 15 with non‐Hodgkin's lymphoma). All the serum samples from the healthy blood donors were negative, In patients with IM and in AIDS‐non Hodgkin's lymphoma (ARNHL), PCR was strongly positive in leukocytes (>2, 000 genome equivalents/l 04 cells), which was correlated with detectable amounts of EBV DNA in serum. The overall positivity rate of PCR in serum was 58.8%, 68%, and 73% of cases for non‐IM primary EBV infections, IM, and ARNHL, respectively. In two cases of EBV primary infection, the viral DNA was detected in serum, respectively 1 month and 2 months before IgM positivity and IgG rise. In one case of ARNHL followed up for several months, PCR (viral load of 2, 000 genome equivalents/104 cells) became positive concurrently with appearance of lymphoma. In immunocompromised individuals, PCR EBV, if carried out in larger prospective studies, could be considered as a tumor marker, useful for predicting EBV‐driven lymphoma and follow‐up therapy. © 1995 Wiley‐Liss, Inc.
Molecular and serologic analysis in the transmission of the GB hepatitis agents
Authors: Schlauder George G. , Dawson George J. , Simons John N. , Pilot‐Matias Tami J. , Gutierrez Robin A. , Heynen Cynthia A. , Knigge Mark F. , Kurpiewski Gretchen S. , Buijk Sheri L. , Leary Thomas P. , Muerhoff A. Scott , Desai Suresh M. , Mushahwar Isa K.
Published: March 2007

Two flavivirus‐like genomes have recently been cloned fron infectious tamarin (Saguinus labiatus) serum, derived from the human viral hepatitis GB strain, which is known to induce hepatitits in tamarins. In order to study the natural history of GB infections, further transmission studies were carried out in tamarins. Reverse‐transcription‐polymerase chain reaction and enzymelinked immunosorbant assays were developedfor the detection of RNA and antibodies associated with the two agents, GB virus‐A and GB virus‐B. The infectivity of both of these agents was demonstrated in tamarins to be filterable through a 0.1 μm filter. Tow distinct genomes were identified in the serum of eight of the infected tamarins, while in four tamarins, the genomes were detected independently of each other. Although specific antibodies to the GB virus‐B epitopes were detected in the serum of animals inoculated with both agents or GB virus‐B alone, antibodies to putative epitopes specific to GB virus‐A were not detected in any of the animals. All tamarins inoculated with serum containing GB virus‐B exhibited an elevation in liver enzyme levels after inoculation. Elevations of serum liver enzyme levels did not occur when GB virus‐A was the only agent detected in the serum. Infection with the original infectious tamarin inoculum conferred protection from reinfection with GB virus‐B but not with GB virus‐A. © 1995 Wiley‐Liss, Inc.
Quantitative assessment of IgM antibodies towards an immunodominant B‐cell epitope within the preS2 domain of HBV in the natural course and during combined prednisone/interferon alpha 2b treatment of chronic hepatitis B virus infection
Authors: Fei Guo‐Zhong , Sylvan Staffan P. E. , Hellström Ulla B. , Yao Guang‐Bi
Published: March 2007

A direct binding enzyme‐linked immunosorbent assay (ELISA) was established for quantitative determination of serum IgM antibodies towards a synthetic peptide corresponding to a selected segment (14‐21) of the preS2‐gene product containing an immunodominant linear B‐cell epitope. The prevalence of IgM anti‐preS2 (14‐21) antibody titers >1, 000 for hepatitis B e antigen (HBeAg)‐positive patients with chronic hepatitis B virus (HBV) infection was 38% (22/58) and 10% (2/21) for HBeAg‐negative subjects (P< 0.005).

IgM anti‐preS2 (14‐21) reactivity was detected during the clinical course of chronic HBV infection and IgM anti‐peptide antibody titers declined and disappeared before spontaneous HBe/anti‐HBe seroconversion.

Recombinant interferon (IFN)‐alpha 2b with an antecedent short course of corticosteroids was administered to eight Chinese patients with chronic HBV infection. The IgM anti‐preS2 (14‐21) reactivity was monitored consecutively during treatment and patients were followed for more than 1 year. A close association between the presence of pretreatment IgM anti‐preS2 (14‐21) in serum and the capacity to respond favorably to the combined prednisone/IFN‐alpha 2b therapy was detected. The IgM anti‐preS2 (14‐21) titers decreased during treatment with subsequent loss of detectable antibodies 8‐16 weeks after the initiation of therapy. This decrease was concomitant with an alanine aminotransferase (ALT) augmentation preceding the disappearance of HBV‐DNA and anti‐HBe seroconversion. Long‐term remission was not observed in treated patients who lacked detectable levels of pretreatment IgM anti‐preS2 (14‐21) in the circulation.

These results demonstrate that a substantial cohort of HBeAg‐positive chronic hepatitis B pa‐tients have IgM antibodies towards a synthetic B‐cell epitope representing a dominant antibody binding site within the envelope of HBV. They indicate that IgM anti‐preS2 (14‐21) titers reflect the immunosuppressive effect on IgM secretion during the combined prednisone/IFN‐α 2b therapy. Further studies are needed to determine the usefulness of a quantitative assay for IgM anti‐preS2 (14‐21) measurements in predicting the response to treatment with interferon alone or preceded by a short course of prednisone in patients with chronic hepatitis B. © 1995 Wiley‐Liss, Inc.
Human B‐cell epitopes of puumala virus nucleocapsid protein, the major antigen in early serological response
Authors: Vapalahti Olli , Kallio‐Kokko Hannimari , Närvänen Ale , Julkunen Ilkka , Lundkvist Åke , Plyusnin Alexander , Lehvaslaiho Heikki , Brummer‐Korvenkontio Markus , Vaheri Antti , Lankinen Hilkka
Published: March 2007

Puumala virus (PUU) is a member of the Hantavi rus genus in the family Bunyaviridae and the etiologic agent of nephropathia epidemica (NE), a form of haemorrhagic fever with renal syndrome (HFRS). In this study we compared the immunofluorescence patterns of NE sera and antibodies raised against recombinant PUU proteins and confirm that the nucleocapsid protein is the major target in the early IgG response of NE patients and provides the molecular basis for simple and rapid differentiation between acute illness and old immunity by granular vs. diffuse fluorescence staining in the indirect immunofluorescence test. The differential kinetics of B‐cell responses to PUU nucleocapsid vs. envelope proteins was emphasized further by the end‐point titres of IgG antibodies to N, G1 and G2 proteins in NE patients. The granular fluorescence correlated with low IgG avidity in 99.8%. and diffuse fluorescence with high avidity in 100% of 617 NE sera studied. Epitope scanning with overlapping 14‐mer peptides covering the whole nucleocapsid protein by a shift of 3 amino acids revealed six major antigenic epitopes recognized by sera from acute‐phase NE patients. The epitopes clustered mainly in the hydrophilic regions, and two of them in a highly variable region which could probably serve as an antigen to distinguish serologically between infections of closely related hantaviruses, some apparently apathogenic, some causing lethal infections. The anti‐peptide epitope pattern varied between different individuals and a collection of several pinbound peptides was needed to be recognised by most NE sera studied. © 1995 Wiley‐Liss, Inc.
Analysis of molluscum contagiosum virus genomes isolated in Japan
Authors: Nakamura J. , Muraki Y. , Yamada M. , Hatano Y. , Nii S.
Published: March 2007

The genomes of 477 Japanese strains of molluscum contagiosum virus (MCV) were analyzed using an in‐gel digestion method with the restriction enzyme BamHI, and classified into four types, including a newly detected type (MCV type 4). All type 1 (MCV‐1) genomes examined so far in Japan showed a common difference from the genome of the MCV‐1 prototype (MCV‐l p), the type reported to be most prevalent in Europe. The common markers of the variants of MCV‐1 were 24‐kbp fusion fragments generated by the loss of a BamHl site between the D2 and F fragments of MCV‐1 p. These variants of MCV‐1 were classified into three groups (MCV‐lva, MCV‐1vb, MCV‐l vc), with the variability among them being due to additions and losses of BamHl sites located in the right terminus and around the Eand I fragments of MCV‐lva. The restriction map of MCVB was generated and lined up with those of the other types. Cross‐hybridization analysis revealed that the organization of all types of MCV genomes were essentially colinear. Considerable numbers of BamHl restriction sites were conserved between MCV‐2 and 4, indicating a close analogy between them. The overall prevalence of MCV, as shown by the ratios of MCV‐1 (MCV‐1p):MCV‐2:MCV‐3:MCV‐4, was 436(0):13: 24:4. Thus, the molecular epidemiology of MCV in Japan is characterized by the absence of the European prototype of MCV‐1, the exclusive occurrence and abundance of variants of MCV‐1, a greater prevalence of MCV‐3 over MCV‐2, and the presence of MCV‐4. © 1995 Wiley‐Liss, Inc.
Synthetic peptides deduced from the amino acid sequence of Epstein‐Barr virus nuclear antigen 6 (EBNA 6): Antigenic properties, production of monoreactive reagents, and analysis of antibody responses in man
Authors: Falk Kerstin , Linde Annika , Johnson Donald , Lennette Evelyne , Ernberg Ingemar , Lundkvist Åke
Published: March 2007

Studies on the antibody responses to various Epstein‐Barr virus (EBV) antigens have been instrumental in the understanding of the seroepidemiology and diagnosis of this viral infection and the subsequent carrier state. While antibodies to the viral capsid antigen (VCA), early antigen (EA), and nuclear antigens 1 and 2 (EBNA 1 and 2) have been well characterized, the antibody response to the other nuclear antigens is not well understood. EBNA 6 is expressed by lymphoblasts during acute EBV infection and may be an important antigen for diagnosis and evaluation of the immune response. In order to analyze the antibody response to EBNA 6, ten peptides (20–21 amino acids), deduced from the EBNA 6 coding region, were synthesized and evaluated for antigenicity by ELISA. One peptide (p‐63; PA‐PQAPYQGYQEPPAPQAPY) derived from the amino acid repeats showed the highest specific reactivity with human sera. This peptide was evaluated further for detection of human EBNA 6‐reactive antibodies. Forty‐two of forty‐nine (86%) EBV‐seropositive healthy donors had p‐63‐ specific IgG reactivity, while none of 50 EBV seronegative patients reacted with the p‐63 peptide. Twenty‐two of fifty‐one (43%) patients with ongoing primary EBV infection had detectable p‐63‐specific IgG. Serum samples drawn sequentially from patients during and after primary EBV infection revealed an increase in p‐63‐reactive IgG over time. A similar pattern was found for reactivity with an EBNA I‐specific peptide (p‐107), in contrast to the EBNA 2 (polyproline) response, which decreased over time. Some EBV seropositive individuals who had no detectable IgG against peptide p‐63 did have antibodies against the native EBNA 6 by anticomplement immunofluorescence to EBNA 6 transfected cells. Rabbit antiserum raised against p‐63 reacted specifically with native EBNA 6 by an immunofluorescence assay and by immunoblotting, indicating the EBNA 6‐specific antigenicity of the peptide. Thus, the peptide p‐63 derived from the amino acid repeats of the EBNA 6 coding region constitutes a predominant, although not exclusive, epitope in the EBNA 6 antibody response. © 1995 Wiley‐Liss, Inc.
Cross‐challenge studies in rhesus monkeys employing different Indian isolates of hepatitis E virus
Authors: Arankalle Vidya A. , Chadha Mandeep S. , Chobe Leenata P. , Nair Rekha , Banerjee Kalyan
Published: March 2007

The aim of this study was to determine if rhesus monkeys infected with one isolate of hepatitis E virus (HEV) were immune to subsequent challenge with other isolates of the virus. Three epidemic and one sporadic Indian HEV isolates were employed in the study. The interval between primary inoculation and challenge varied from 1 year and 6 months to 2 years and 9 months. Evidence of HEV infection was ascertained by rise in serum alanine transaminase (ALT) levels and/or seroconversion to antibodies to HEV (anti‐HEV), and the presence of HEV‐RNA in the bile or faeces of the infected monkeys. No evidence for multiplication of virus in monkeys challenged with different HEV isolates was obtained. These results show that immunity generated by one isolate of HEV protects against different isolates of hepatitis E virus. © 1995 Wiley‐Liss, Inc.
Detection of HCV RNA in serum and seminal fluid from HIV‐1 co‐infected intravenous drug addicts
Authors: Fiore Ramon José , Potenza Domenico , Monno Laura , Appice Amelia , Distefano Mariantonietta , Giannelli Anna , Lagrasta Lidia , Romanelli Chiara , Dibari Cesare , Pastore Giuseppe
Published: March 2007

The presence of hepatitis C virus (HCV) RNA in serum and seminal fluid was investigated in eleven drug addicts coinfected with HIV‐1 and HCV. Serum and seminal fluid were taken from each patient at the same time point. HCV RNA was found in ten of the eleven serum samples tested, but only in one of the semen samples. No relationship was observed between CD4 cell counts, the stage of HIV infection, extent of liver damage and the presence of HCV RNA in serum and semen. The results indicate that HCV is not usually present in the semen and provide further evidence against sexual transmission as an important mode of transmission of HCV infection. © 1995 Wiley‐Liss, Inc.
Demonstration of either endogenous recurrence or exogenous reinfection by resiriction endonuclease cleavage analysis of herpes simplex virus from patients with recrudescent genital herpes
Authors: Sakaoka Hiroshi , Aomori Tsugumitsu , Gouro Tsutomu , Kumamoto Yoshiaki
Published: March 2007

By using restriction endonuclease (RE) cleavage analysis, either endogenous recurrence or exogenous reinfection of herpes simplex virus (HSV) was clarified in 45 male and 20 female subjects with recrudescent genital herpes. All of the plural (two to ten) isolates from 63 (205 isolates) out of 65 subjects (97%) were HSV‐2. Two isolates from only one of 65 subjects (1.5%) were HSV‐1, and they showed the same RE profile. In addition, an HSV‐1 and five HSV‐2 isolates were obtained from the remaining one female patient (1.5%), indicating that an exogenous HSV‐1 strain has been reinfected during HSV‐2 recrudescences. HSV‐2 isolates were furthermore classified into genotypes of HSV‐2 using 16 different RE markers with five REs. Two hundred and ten HSV‐2 isolates from 64 subjects were classified into 27 distinct genotypes, in which some predominant genotypes and seven new genotypes were found. Plural HSV‐2 isolates obtained from 63 out of 64 subjects, including one subject from whom an HSV‐1 and five HSV‐2 strains were isolated, were classified into the same genotypes, indicating that they may be regarded as recrudescent genital herpes by the reactivation of the same endogenous strain. However, the RE profiles of two HSV‐2 strains from the remaining one subject were different. Thus, it was finally found that only two out of 65 subjects (3%) were reinfected with exogenous strains. These results lead to the conclusion that almost all recrudescent genital herpes are due to the reactivation of an initially infected HSV‐2 strain, and are occasionally due to reinfection with distinct HSV strains of either the same or a different type. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus RNA in peripheral blood mononuclear cells: Relation with response to interferon treatment
Authors: Taliani Gloria , Badolato Maria Coneetta , Lecce Rosina , Poliandri Giuseppina , Bozza Alessandro , Duca Franca , Pasquazzi Caterina , Clementi Carlo , Furlan Caterina , De Bac Carlo
Published: March 2007

The polymerase chain reaction (PCR) was used to investigate the presence of positive and negative hepatitis C virus (HCV) RNA strands in serumand peripheral blood mononuclear cells (PBMC) of 20 patients with histologically proven HCV‐ related chronic liver disease. All patients completed a course of interferon (IFN) treatment (6 MU of IFN‐α2b three times a week for 24 weeks) and were followed‐up for 12 months after treatment was discontinued. Pre‐treatment, end‐treatment and 6‐month follow‐up serum and PBMC samples were examined. At enrollment, the positive strand of HCV‐RNA was detected in serum of 18 patients (90%), the negative strand in none. Positive‐stranded HCV‐RNA was detected in PBMC of 15 patients (75%), 13 of whom also had detectable levels of negative‐stranded HCV‐RNA in PBMC. By the end of the treatment, 12 patients (60%) were responders. The pre‐treatment HCV infection of PBMC, indicated by the presence of both RNA strands, was found in 8 (66.7%) responders compared to 5 (62.5%) non‐responders (P=n.s.). End‐treatment loss of PMBC HCV‐RNA correlated significantly with the response since it occurred in all responders compared to 2 non‐responders (P=0.02). However, end‐treatment‐negative serum and PBMC HCV‐RNA did not predict the occurrence of a sustained response, which was observed at month 12 in 5 of 12 responders (P= n.s.). On the other hand, the persistent absence of HCV RNA in serum and PBMC at the end of the 6‐month follow‐up was significantly associated with the occurrence of a sustained response (P<0.0001). © 1995 Wiley‐Liss, Inc.