bluuebunny/crossref_metadata_2025 huggingface dataset:
Hepatitis E virus infection in hemophiliacs
Authors: Barzilai Asher , Schulman Sam , Karetnyi Yuory V. , Favorov Michael O. , Levin Edit , Mendelson Ella , Weiss Peretz , Fields Howard A. , Varon David , Martinowitz Uri
Published: March 2007

Israel is endemic for hepatitis E virus (HEV), the causative agent of enteric non‐A, non‐B hepatitis. Transmission is via the feco‐oral route but the possibility of transmission through blood transfusion has been raised. This question was addressed by examining sera from 188 hemophilic patients in Israel. screening was performed with an enzyme immunoassay (EIA) for antibody against hepatitis E virus (anti‐HEV) and confirmed with a neutralization test. Sixteen patients (9%) were seropositive for anti‐HEV. A statistically significant difference was not found between the seroprevalence in this group and that of a healthy Israeli control population, matched for sex and age. The anti‐HEV‐seropositive hemophiliacs had the same seroprevalence of antibodies to hepatitis B and C virus and to HIV and the same number of cases with chronic hepatitis as among the anti‐HEV‐seronegative patients. The seroprevalence of antibodies to hepatitis A virus (anti‐HAV) was, on the other hand, higher in the anti‐HEV‐seropositive group. This study indicates that HEV is not transmitted by cryopre‐cipitate or lyophilized factor concentrates. High prevalence of coinfection with hepatitis A supports our conclusion that HEV infection in Israeli hemophiliacs was due mainly to feco‐oral transmission. © 1995 Wiley‐Liss, Inc.
Mutations of some critical amino acid residues in the hepatitis B virus surface antigen
Authors: Bruce Sandra A. , Murray Kenneth
Published: March 2007

Amino acid substitutions at several positions in the surface antigen (HBsAg) of hepatitis B virus (HBV) in natural isolates and the products of recombinant DNA molecules have identified important residues for cross‐reaction with specific antibodies (anti‐HBs) and the induction of antibodies with certain serological specificities. In a further group of mutants described here, cysteine residues in a region believed to be significant of the a epitope have been changed to serines. Of the three adjacent cysteine residues at positions 137, 138 and 139, mutation of either of the flanking residues reduced cross‐reactivity with polyclonal anti‐HBs, while alteration of the central residue was relatively well‐tolerated. Mutation of cysteine 149 to serine or of glycine 145 to arginine (imitating naturally occurring mutants), lysine, or glutamatic acid all led to loss of cross‐reactivity with polyclonal antisera. © 1995 Wiley‐Liss, Inc.
Semiquantitative anti‐HBc IgM detection in children with chronic hepatitis B: A long‐term follow‐up study
Authors: Gaeta G. B. , Nardiello S. , Pizzella T. , Russo G. , Maisto A. , Sardaro C. , Galanti B. , Giusti G.
Published: March 2007

Serum anti‐HBc IgM titres were monitored monthly by a semiquantitative method in 14 children with HBeAg positive chronic hepatitis B followed up for 18‐65 months. All patients, but one, were treated with alfa‐interferon (IFN) at different times. On the whole, 12 flare‐up episodes were observed and 7 patients cleared HBV‐DNA and seroconverted to anti‐HBe. Seroconversion occurred only in patients with pretreatment anti‐HBc IgM index greater than 0.15 and serum HBV‐DNA concentration below 100 pg/ml; the pretreatment alanine aminotransferase (ALT) value was not predictive of response. Combining anti‐HBc IgM results and serum HBV‐DNA levels observed during the pre‐IFN period allowed a precise identification of patients who were likely to respond to IFN therapy.

Patients who seroconverted to anti‐HBe showed a progressive reduction in serum anti‐HBc IgM titres within 6 months. Interestingly, one child, in whom HBV‐DNA reappeared andwho reconverted to HBeAg 7 months after treatment, showed no anti‐HBc IgM decrease afterthe transient clearance of HBV‐DNA and anti‐HBe seroconversion. Semiquantitative anti‐HBcIgM detection is a useful tool in the decision making process for children with chronic hepatitis B. © 1995 Wiley‐Liss, Inc.
Genetic susceptibility to herpetic encephalitis of inbred rabbits of B/Jas strain
Authors: Seto Akira , Morikawa Shigehiro , Isono Takahiro , Ogawa Katsuhiko , Inubushi Toshiro
Published: March 2007

Inbred rabbits of B/Jas strain were found to be highly susceptible to herpes simplex virus type 1 encephalitis, following i.v. injection of the virus, while Chbb:HM strain rabbits were not susceptible. The susceptibility trait seemed to be inherited recessively, involving multiple genes, because (B/Jas x Chbb:HM)Fl hybrids were as resistant as Chbb:HM rabbits, and because more than 90% of backcrosses of (B/Jas x Chbb: HM)FI to B/Jas were resistant to viral inoculation. The encephalitis in B/Jas rabbits resembled human herpes simplex encephalitis, in that the temporal lobe as well as the brain stem were affected preferentially, leading to the development of various types of seizures, such as circling, loss of balance leading to a fall, and tonic and clonic convulsions. The disease could be diagnosed by magnetic resonance imaging (MRI) analysis before onset of seizures, and diseased rabbits showed a marked lymphopenia at onset of seizures. © 1995 Wiley‐Liss, Inc.
Oral brivudin vs. intravenous acyclovir in the treatment of herpes zoster in immunocompromised patients: A randomized double‐blind trial
Authors: Wutzler Peter , De Clercq Eric , Wutke Klaus , Färber Inge
Published: March 2007

The efficacy of oral brivudin vs. intravenous acyclovir was compared in a randomized multicentered study under double‐blind conditions using the double‐dummy technique. Forty‐eight patients with a herpes zoster rash less than 72 hours in duration were entered in the study. Brivudin was given as one 125‐mg tablet every 6 hours. Acyclovir was infused over 1 hour at a dose of 10 mg/kg every 8 hours. Treatment was continued for 5 days. There was no significant difference between the treatment groups when analyzed in terms of new lesion formation, increase in the area of rash within the primary dermatome, cutaneous dissemination, and affection of mucous membranes or visceral organs. Both treatment regimes were also equally effective in the time to full crusting of lesions. Oral brivudin and intravenous acyclovir were well tolerated by most patients. There was no need to interrupt the treatment in any case. As effective as intravenous acyclovir in the treatment of herpes zoster, oral brivudin offers the potential for outpatient treatment of herpes zoster in immunocompromised patients. © 1995 Wiley‐Liss, Inc.
Fatal illness associated with a new hantavirus in Louisiana
Authors: Khan Ali S. , Spiropoulou Christina F. , Morzunov Sergey , Zaki Sherif R. , Kohn Melvin A. , Nawas Soheir R. , McFarland Louise , Nichol Stuart T.
Published: March 2007

A fatal case of hantaviral illness occurred in Louisiana, outside of the range of P. maniculatus, the rodent reservoir for Sin Nombre virus. Hantavirus RNA and antigens were detected in patient autopsy tissues, and nucleotide sequence analysis of amplified polymerase chain reaction (PCR) products identified a newly recognized unique hantavirus, provisionally named Bayou virus. Prominent features of the clinical illness are compatible with hantavirus pulmonary syndrome (HPS), but several features such as renal insufficiency and intraalveolar hemorrhage are more compatible with hemorrhagic fever with renal syndrome (HFRS), a disease associated with Eurasian hantaviruses. © 1995 Wiley‐Liss, Inc.
Hepatitis C virus seroconversion rate in established blood donors
Authors: Atrah H. I. , Hutchinson F. , Gough D. , Ala F. A. , Ahmed M. M.
Published: March 2007

The results of hepatitis C virus (HCV) antibody test of 237, 813 blood donations collected from 143, 815 donors by the West Midlands Blood Transfusion Centre in 1993 were analyzed retrospectively in order to determine the seroconversion rate among established previously anti‐HCV negative donors. Three hundred sixteen (0.22%; 1 in 455) donors were positive by the enzyme linked immunosorbent assay (ELISA) screening test and 34 (0.024%; 1 in 4, 230) donors were positive by ELISA and the Recombinant Immuno Blot Assay (RIBA). Three donors previously negative for HCV antibody reacted positively by both tests. The annual seroconversion rate was calculated as one in 35, 937 donors. This figure argues against limitation of HCV antibody screening to new blood donors. A further 45 donors negative on previous screening reacted positively by ELISA and were indeterminate by RIBA. Unexpectedly, lapsed blood donors first tested for HCV antibody in 1993 had high positive reaction rates by ELSA and RIBA, which was significantly (P < 0.001) higher than those of new donors. RIBA‐positive reaction rate among ELISA‐positive donors was significantly higher amongst males than females (P < 0.0011. © 1995 Wiley‐Liss, Inc.
Prospective study on the development of antibodies against human papillomavirus type 6 among patients with condyloma acuminata or new asymptomatic infection
Authors: Wikström A. , van Doornum G. J. J. , Kirnbauer R. , Quint W. G. V. , Dillner J.
Published: March 2007

The development over time of antibodies to human papillomavirus type 6 (HPV 6) was studied prospectively in a group of 26 condyloma patients receiving multiple treatments, as well as in 17 healthy subjects with a new HPV 6 infection. The condyloma patients provided serum samples during 2–12 visits over a period of 1–20 months, and the levels of IgA, IgG and IgM antibodies to HPV 6, 11 and 16 capsids, and to HPV 6‐derived peptide antigens were determined. Among the capsid antibodies, the IgG and IgA responses against HPV 6 were the most prominent, whereas IgM was detected only in a few patients. During follow‐up, seroconversion to HPV capsids was found in six patients and antibody peaks disappeared during follow‐up in nine patients. There was no association between serology and cure of the wart.

One hundred twenty subjects who reported multiple sexual partners, but found negative for HPV DNA at multiple anogenital and oral sites, provided serum samples and swabs for HPV DNA analysis at 2–7 visits during 4–74 months. Seventeen subjects acquired HPV 6 DNA and were included in the study. Serum IgA, IgG and IgM antibodies to HPV 6 capsids were induced concomitantly with the acquisition of viral DNA (P = 0.02, 0.002 and 0.006, respectively), and declined again when the viral DNA was no longer detectable.

In conclusion, antibodies to HPV 6 did not predict the clinical course of condylomatous disease, but appear to be useful mainly as epidemiological markers of viral exposure. © 1995 Wiley‐Liss, Inc.
Rapid genotyping of hepatitis C virus RNA‐isolates obtained from patients residing in Western Europe
Authors: Kleter G. E. M. , van Doorn L. J. , Stuyver L. , Maertens G. , Brouwer J. T. , Schalm S. W. , Heijtink R. A. , Quint W. G. V.
Published: March 2007

Two rapid genotyping methods for hepatitis C virus (HCV), the line probe assay (Inno‐LiPA) and the subtype‐specific core amplification system [Okamoto et al., (1992b) Journal of General Virology 73:673‐679], were applied to 58 HCV isolates which were typed as type 1 (n=37) and type 2 (n=21) by sequence analysis of the 5′ untranslated region (5′UTR). The line probe assay targets the 5′UTR and recognized 12 subtype 1a, 25 subtype 1b, 18 subtype 2a, 2 subtype 2b and 1 subtype 2d in accordance with sequence analysis of this region. Subtype‐specific core amplification revealed 7 discrepancies among the 37 type 1 isolates when compared to LiPA. A different subtype was observed in 3 isolates (la versus 1b), 2 isolates remained untyped and 2 isolates showed a coinfection of subtype la and 1b. The first 5 discrepancies were confirmed by sequence analysis of the core region whereas the coinfection could not be confirmed. Of the 21 type 2 isolates only one could be typed by subtype‐specific core amplification. HCV RNA was detected in all 21 cases after the general first round of polymerase chain reaction (PCR). Direct sequencing of the core region indicated sequence variation as a source of failure.

It is concluded that LiPA results are conclusive for typing of HCV. However, LiPA is hampered occasionally for subtyping by lack of subtype specific sequence variation in 5′UTR. Subtyping results by subtype‐specific core amplification were accurate. However, it seems that this assay is not suitable for the identification of genotype 2 isolates that circulate in patients living in Western Europe. © 1995 Wiley‐Liss, Inc.
Quantitation of human cytomegalovirus genomes in the brain of AIDS patients
Authors: Kühn J. E. , Wendland T. , Eggers H. J. , Lorentzen E. , Wieland U. , Eing B. , Kiessling M. , Gass P.
Published: March 2007

Human cytomegalovirus (HCMV) is one of the major pathogens causing neurologic disease in the immunocompromised host. A competitive nested polymerase chain reaction (PCR) was used to determine DNA load, distribution, and sequence variability of HCMV genomes in the brain of AIDS patients with and without HCMV encephalitis confirmed by histology and immunocytochemistry. By quantitative PCR, HCMV genomes were found to be distributed diffusely in the central nervous system (CNS) of all five patients with histologically proven HCMV encephalitis, but also in the brain of five of eight AIDS patients without neuropathological evidence of HCMV encephalitis. The viral DNA load in cases with HCMV encephalitis was increased 10‐ to 1, 000‐fold as compared to patients without evidence of encephalitis. A viral load above 6, 000 copies HCMV/106 copies β‐globin was found to be highly suggestive for HCMV encephalitis. Characterization of PCR products by temperature gradient gel electrophoresis (TGGE) and direct sequencing allowed us to detect a sequence variability of the amplified fragment of HCMV glycoprotein B (gB) among different patients, but also among different HCMV foci within the same patient. Furthermore, two of five AIDS patients with HCMV encephalitis most likely experienced double infections with different HCMV strains. The experimental procedure described in this study should also be applicable to the detection of significant HCMV DNA levels in biopsy samples. © 1995 Wiley‐Liss, Inc.
Comparison of human immunodeficiency virus biological phenotypes isolated from cerebrospinal fluid and peripheral blood
Authors: Peeters Martine F. , Colebunders Robert L. , Van Den Abbeele Koen , Nys Patrick C. , Goeman Johan , Colans Philippe , Geerts Yvette , Van der Groen Guido
Published: March 2007

Quantitative human immunodeficiency virus (HIV) cultures were carried out on cerebrospinal fluid (CSF), peripheral blood mononuclear cells (PBMCs), and plasma from patients with HIV in order to compare the infectious HIV load. The HIV strains isolated were studied for syncytiuminducing (SI) capacity, using the MT‐2 cell line, in order to compare the HIV strain phenotype of blood and CSF isolates.

Forty‐two patients with HIV‐1 infection were enrolled in the study, 33 of whom had neurological symptoms and 9 of whom were without neurological symptoms. HIV was isolated from 16 (38%) of the 42 CSF cultures, with a low mean titer of 6.3 ± 3.4 tissue‐culture‐infective doses (TCID) per milliliter. Patients with HIV‐positive CSF culture had a viral load in PBMCs of 40.5 ± 15.5 TCID per 106 PBMC and in plasma of 104.7± 9.3 per milliliter. Two (15%) of the 13 CSF isolates were Sl strains, compared to 17 (56.6%) of the 30 PBMC isolates and 13 (54%) of the 24 plasma isolates (P<0.05). Five of the nine patients from whom CSF and blood strains were obtained had the same viral biological phenotype.

This study suggests that different HIV variants may be found in different body fluids and/or cells. © 1995 Wiley‐Liss, Inc.
Base changes at positions 1014 and 578 of delta virus RNA in greek isolates maintain base pair in rod conformation with efficient RNA editing
Authors: Yang Ailing , Papaioannou Christos , Hadzyannis Stephanos , Thomas Howard , Monjardino John
Published: March 2007

Analysis of delta hepatitis virus (HDV) genomic RNA, derived from Greek patients from an area where HDV infection is associated with low pathogenicity, is described. In all isolates sequenced, which included 18/18 HDV cDNA clones derived from 6 different patients, irrespective of pathogenicity, a base change (T→C) was found in position 1014. No significant differences in editing efficiency were found between isolates from inactive and active forms of the disease, although L‐antigen was present in low to undetectable levels in the serum of 5/6 patients. An additional mutation was identified at position 578 (A→G), which reestablishes the canonical base pair G/C with the mutated 1014 when the genome adopts the “rod‐like” conformation. This finding supports the presence of this genome conformation in vivo and the requirement for the Watson‐Crick base pair 10141/578. A mutation, found at amino acid position 170 (serine → asparaginel, appears to segregate with patients with inactive disease. © 1995 WiIey‐Liss, Inc.
Epidemiologic applications of novel molecular methods to detect and differentiate small round structured viruses (Norwalk‐like viruses)
Authors: Ando Tamie , Jin Qi , Gentsch Jon R. , Monroe Stephan S. , Noel Jacqueline S. , Dowell Scott F. , Cicirello Helen G. , Kohn Melvin A. , Glass Roger I.
Published: March 2007

The molecular epidemiology of a large, multi‐state outbreak of oyster‐associated gastroenteritis [Kohn et al. (1995): Journal of the American Medical Association 273:466–471. Dowell et al. (1995): Journal of Infectious Diseases 171:1497–1503.] was examined using new methods to detect small round structured viruses (SRSVs) by reverse transcription‐polymerase chain reaction (RT‐PCR) and to characterize strains by Southern hybridization and nucleotide sequencing of 81‐bp of a PCR product amplified from the RNA polymerase gene. Of 37 stool specimens examined from patients in eight clusters of the multi‐state outbreak, 32 (86%) gave RT‐PCR products specific for SRSVs of PI‐A phylogenetic group. Nineteen PCR products from the eight clusters were confirmed to have the identical sequence, indicating that this large outbreak was attributed to a single strain of SRSV. In one of the eight clusters, five (63%) of eight patients had a mixed infection with a second SRSV strain that belonged to P2‐B phylogenetic group. Of 12 specimens from patients in five other outbreaks and one sporadic case which occurred at the same time as the multistate outbreak, 10 (83%) gave products specific for SRSVs representing four phylogenetic groups (PI‐A, PI‐B, P2‐A, and P2‐ B). The sequences of the PI‐A products from two outbreaks and that of the P2‐B product from another outbreak were identical to the P1‐A sequence from the eight clusters and the P2–6 sequence from the one cluster of the multistate outbreak, respectively. These results demonstrate the first application of these methods to enhance our understanding of the molecular epidemiology of SRSVs and provide answers of public health interest that could not have been obtained using classical epidemiologic methods alone. © 1995 Wiley‐Liss, lnc.

This article is a US Government work and, as such, is in the public domain in the United States of America.
Large‐scale study suggests no direct link between human herpesvirus‐6 and primary Sjögren's syndrome
Authors: Ranger‐Rogez S. , Vidal E. , Labrousse F. , Riche A. , Vidal J. , Collineau M. , Liozon F. , Denis F.
Published: March 2007

The aim of this study was to evaluate the role of human herpesvirus‐6 (HHV‐6) in the development of primary Sjögren's syndrome (PSS). Serum HHV‐6 antibody levels, as measured by immunofluorescence assay (IFA) and the prevalence of HHV‐6 DNA in peripheral blood mononuclear cells (PBMCs) determined by poly‐merase chain reaction (PCR), were studied in 49 PSS patients and 50 control subjects, all in‐patients in the University Hospital Internal Medicine ward, Limoges, France. In addition, portions of labial salivary gland were obtained from 34 patients and 15 controls, the presence of viral DNA being detected by the same PCR technique. The results were then compared with clinical observations of systemic disease manifestations in patients and a histological study of salivary gland involvement. No significant difference in HHV‐6 seroprevalence was found between control subjects (50.0%) and patients (63.3%) nor was there any statistically significant difference between patient and control groups for total viral DNA in PBMCs (22.4%, 12.0%) and salivary glands (8.8%, 6.6%). Analysis of clinical and histological data revealed no detectable correlation between disease severity and viral involvement. Tests for HHV‐6A and HHV‐6B proved positive in patient and control groups, HHV‐6B being the most frequently encountered type in both groups. In conclusion, the results of this large‐scale trial does not confirm the suspected direct role of HHV‐6 in the etiology of PSS. © Wiley‐Liss, Inc.
Quantitation of anti‐hepatitic C virus antibodies in interferon‐treated patients by S29‐1/S4 ELISA
Authors: Sato Atsushi , Nakamura Haruji , Sho Yukihiko , Arima Terukatsu
Published: March 2007

In 23 patients with chronic hepatitis C who have undergone interferon (IFN) treatment, quantitation of anti‐hepatitis C virus IgG antibodies by a second‐generation assay S29‐1/S4 ELISA [Sato et al. (1993): Microbiology and Immunology 37: 295‐304]. (anti‐S29‐1/S4) was compared with that of anti‐HCV core IgG antibodies (C22‐3, anti‐core) and the presence of viral RNA confirmed by the reverse transcription‐nested polymerase chain reaction (RT‐nested PCR). In 12 complete responders with loss of HCV‐RNA and normal alanine aminotransferase (ALT) levels at 6 months after therapy, IgG antibodies quantified by a second‐generation assay have decreased significantly at the end of treatment (P < 0.05). Further significant reduction of anti‐S29‐1/S4 titers was observed at 6 months after therapy (P < 0.01) as well as anti‐core antibodies (P < 0.01). On the other hand, in 11 non‐responders with persistent or intermittent viremia at 6 months after therapy, no significant change in the level of anti‐S29‐l/S4 titers was observed, whereas anti‐core titers have decreased at the end of therapy (P < 0.01). In contrast, both levels of anti‐S29‐1/S4 and anti‐core have increased at 6 months after treatment (P < 0.05). It is concluded that, although it appears difficult to monitor the effect of virus clearance during IFN therapy, changes in anti‐S29‐l/S4 titers after IFN treatment correlate with virus clearance and with anti‐core titers. © Wiley‐Liss, Inc.
Quantitative measurement of HBeAg in chronic hepatitis B: A comparison between a radioimmunoassay a fluorescence ELISA and a chemiluminescence ELISA
Authors: Heijtink R. A. , Snobl J. , Kruining J. , Kerkhof‐Los C. , de Man R. A. , Janssen H. L. A. , Schalm S. W.
Published: March 2007

The presence of the hepatitis B e antigen (HBeAg) in peripheral blood of chronic hepatitis B patients is a widely accepted marker of active replication of the hepatitis B virus. HBeAg determination during interferon therapy is a useful guide for the therapeutic regimen.

The aim of the study was to compare the suitability of an HBeAg radioimmunoassay (RIA, Abbott Laboratories, North Chicago, IL, USA), the IMx‐HBeAg assay (IMx, Abbott Laboratories) and the HBeAg/anti‐HBe Amerlite assay (Amerlite, Johnson & Johnson Clinical Diagnostics, Cardiff, UK) for semiquantitative monitoring of HBeAg during therapy. HBeAg levels in serum samples obtained before and during interferon therapy were measured using an in‐house standard calibrated against the Paul Ehrlich Institute HBeAg reference preparation (PEI standard).

When serial dilutions of pretreatment serum samples were assayed by the three methods, radioimmunoassay was found to be highly sensitive although it had a very limited working range (0.5 to 12 PEI U/ml). A broader linear working range was observed for Amerlite (0.5 to 50 PEI U/rnl) and the IMx assay (0.5 to 100 PEI U/ml). The intra‐assay and interassay variations did not differ significantly. Since the IMx assay was less susceptible to sample variation and had a broad working range, semiquantitative measurement of HBeAg in one diluted and one undiluted sample by this assay may justifiably be introduced as routine procedure. Routine semiquantitative HBeAg measurement may improve individual dose adjustments and thus the success of interferon therapy. © Wiley‐Liss, Inc.
Experimental myocarditis induced by two different coxsackievirus B3 variants: Aspects of pathogenesis and comparison of diagnostic methods
Authors: Arola Anita , Kalimo Hannu , Ruuskanen Olli , Hyypiä Timo
Published: March 2007

The ability of two coxsackievirus B3 (CBV3) variants to induce myocarditis in BALB/c mice was studied and plaque‐forming assay, polymerase chain reaction (PCR), in situ hybridization, and immunohistochemistry were compared for detecting viruses and viral components in the myocardium. The virological findings were related to histopathologic and ultrastructural changes in the myocardium. CBV3‐W induced severe myocarditis characterized by massive myocyte necrosis. Widely distributed myocyte damage clearly preceded modest inflammatory infiltrates in the myocardium. In contrast, CBV3‐MI induced mild myocardial injury. Both variants caused fulminant pancreatitis with nearly complete necrosis of the exocrine pancreas. CBV3 RNA was identified by PCR in the myocardium of CBV3‐W‐infected mice until the end of the follow‐up period of 14 days. Moreover, semiquantitative results were obtained when the PCR/ hybridization results were analyzed by a phosphor imaging system. lmmunohistochemistry and in situ hybridization from formaldehyde‐fixed, paraffin‐embedded specimens were highly similar in detecting viral components during the early stages of the myocardial injury. The results indicate that: (i) direct viral damage plays an essential role in acute murine CBV3‐induced myocarditis, (ii) PCR appears a useful and sensitive diagnostic method in acute myocarditis, and (iii) immunohistochemistry as a specific and relatively rapid method might be practicable also in studying the early stages of acute myocarditis from archival clinical material. © Wiley‐Liss, Inc.
Biological phenotypes of HIV‐1 subtypes A and B strains of diverse origins
Authors: Nkengasong John N. , Peeters Martine , Zhong Ping , Willems Betty , Janssens Wouter , Heyndrickx Leo , Fransen Katrien , Ndumbe Peter M. , Gershy‐Damet Guy‐Michel , Nys Patrick , Kestens Luc , Piot Peter , van der Groen Guido
Published: March 2007

The identification of specific biologic phenotypic traits that can be correlated with different HIV‐1 genetic subtypes was sought. The genetic subtypes were determined by either sequencing (Cameroonian strains, n = 18) or by the heteroduplex mobility assay (HMA) (Belgian strains, n = 21 and lvorian strains, n = 25). Seventeen (81%) of the 21 Belgian isolates belonged to subtype B and 4 (17%) were subtype A strains. Subtype A variants were predominant in the two African countries studied; 11 (61%) of 18 strains from Cameroon and 23 (92%) of 25 strains from the Ivory Coast. Of the 64 isolates, 38 (58%) and 19 (29%) belonged to subtypes A and B, respectively. No significant difference was observed for biological phenotypes (slow/low and rapid/high) of both genetic subtypes. In symptomatic individuals, however, a significantly higher number of subtype B isolates were of rapid/high phenotype, compared with subtype A (5 of 10; 50%) vs. 2 of 22; 9%), respectively; X2 = 6.7, P = 0.02). The findings suggest that overall HIV‐1 isolates belonging to genetic subtype B are not distinguishable from subtype A variants on the basis of their biological phenotypes. Syncytium‐inducing variants were less prevalent regardless of the geographic origin of the isolates. © Wiley‐Liss, Inc.
Significance of anti‐hepatitis C virus core IgM antibodies in patients with chronic hepatitis C
Authors: Pawlotsky Jean‐Michel , Darthuy Françoise , Rémiré Jocelyne , Pellet Claire , Udin Laure , Stuyver Lieven , Roudot‐Thoraval Franqoise , Duvoux Christophe , Douvin Catherine , Mallat Ariane , Mavier Jean‐Phillippe , Métreau Jean‐Michel , Duval Jean , Dhumeaux Daniel
Published: March 2007

Antihepatitis C virus (HCV) IgM antibodies were found in patients with both acute and chronic hepatitis C. The aims of the study were to determine the significance, in terms of liver disease and virological parameters, of anti‐HCV core IgM antibodies in the serum of patients with chronic hepatitis C, and the possible relationship between the presence of these antibodies before treatment and biochemical and virological responses to interferon therapy. Sixty‐one patients with chronic hepatitis C were studied. Tests for serum anti‐HCV core IgM antibodies were carried out before treatment. The patients received 3 mega units of interferon alpha‐2a subcutaneously thrice weekly for at least 3 months (6months when alanine aminotransferase activity was normal at month 3). A biochemical response to interferon therapy was defined as normal alanine aminotransferase activity at the end of treatment (month 6: biochemical response) and 6 months later (month 12: sustained biochemical response). A sustained virological response was defined as serum HCV RNA negativity by a polymerase chain reaction‐based detection method (PCR) in patients with normal alanine aminotransferase at month 12. Anti‐HCV core IgM antibodies were detected in 28 of the 61 patients (46%). The prevalence of these antibodies was significantly higher in patients infected with HCV genotype 1 (including subtypes la and 1b) than in patients infected with other genotypes (including 2a and 3a) (57% vs. 17%; P < 0.01). No significant difference was found between IgM‐positive and IgM‐negative patients as regards the mean age, sex ratio, serum alanine aminotransferase and gamma‐glutamyl transpeptidase activities, the prevalence of cirrhosis in liver biopsy specimens, detection of HCV RNA by PCR, and quantitation by branched DNA assay. At month 6 of interferon therapy, normal alanine arninotransferase activity was significantly more frequent in lgM‐negative than in IgM‐positive patients (52% vs. 21%, respectively; P < 0.02). At month 12, normal alanine aminotransferase activity and PCR negativity were significantly more frequent in IgM‐negative than in IgM‐positive patients (18% vs. Ooh, P < 0.04). It is concluded that antiHCV core IgM antibodies in serum are significantly more frequent in patients infected by HCV type 1 than by other types. This suggests that their overall prevalence in patients with chronic hepatitis C in industrialized countries, where HCV type 1 accounts for the majority of infections, would be of the order of 50%, that antiHCV core IgM antibodies are not associated with characteristic features of liver disease, and that their presence before treatment is associated with a failure of interferon alpha therapy to clear the virus. © Wiley‐Liss, Inc.
Selection of a precore mutant after vertical transmission of different hepatitis B virus variants is correlated with fulminant hepatitis in infants
Authors: Bahn Andrew , Hilbert Katja , Martiné Ursula , Westedt Jens , Weizsäcker Friedrich V. , Wirth Stefan
Published: March 2007

The incidence of perinatal transmission of hepatitis B virus (HBV) depends on the HBeAg/anti‐HBe status of the mother. While children of HBeAg‐positive mothers have a 90% probability of acquiring a chronic hepatitis B virus carrier state, babies of anti‐HBe‐positive mothers are more likely to develop fulminant hepatitis within the first 3 to 4 months of life. There is evidence that precore (pre‐C) mutations of the HBV can be associated with fulminant hepatitis. The pre‐C region was therefore examined in sera from nine infants with fulminant hepatitis after vertical transmission, one HBeAg‐positive and seven anti‐HBe‐positive mothers by polymerase chain reaction (PCR) and direct sequence analysis. In five mother/infant pairs the virus populations were characterized in addition by analysing clones of the amplified products. All mothers were infected with two or four variants of HBV with mutations at different positions of the preC genome including position 1896, which results in a stop codon. While the precore stop codon was detected in a portion of the virus populations of the HBeAg‐positive and of four anti‐HBe‐positive mothers the dominating viral strain was represented by the wild type virus in three. In contrast, the virus populations of all babies showed the 1896 precore variant as the prevalent virus strain during the phase of active disease. In the surviving baby only wild type sequences were detected after recovery. Subtype ayw was found in all mothers and infants and adw2 was present in three mothers and in the surviving child.

The findings suggest that all mothers carried a wild type HBV population with a certain number of different HBV variants. After transmission of the mixed virus population a selection process was started in the baby. The association of subtype ayw with the precore mutations and with the fatal outcome of the hepatitis B might be the result of a directed selection of this variant with a particular advantage in the viral life cycle. © Wiley‐Liss, Inc.
Homotypic immune response to primary infection with rotavirus serotype G1
Authors: Rojas A. M. , Boher Y. , Guntinas M. J. , Pérez‐Schael I.
Published: March 2007

Some aspects of rotavirus humoral immunity were assessed on the basis of distinguishing se‐rotype‐specific specificities (VP4/VP7) by using rotavirus reassortants, human and animal strains in neutralization assays in serum samples obtained during the acute phase, and 1, 6 and 12 months after primary natural infection. In this study, all the infecting virus strains were characterized as G type and some also as P type. Primary natural infection induces a significantly greater homotypic neutralization response than heterotypic response. In addition, there was no significant difference in the number of homotypic or heterotypic responses following reinfection. Transplacentally acquired homotypic antibodies were associated with protection against dehydration during rotavirus gastroenteritis. © Wiley‐Liss, Inc.
Virus‐specific, antibody‐secreting cells during upper respiratory infections
Authors: Mäkelä Mika J. , Nikkari Simo , Meurman Olli , Laine Matti , Arvilommi Heikki
Published: March 2007

The humoral immune response of 18 army recruits with febrile upper respiratory infection (URI) was studied by enumerating virus‐specific, antibody‐secreting cells in the peripheral blood. Diagnosis was based on viral antigen detection in nasopharyngeal specimens, virus isolation from throat swabs, or on antibody measurement from paired serum samples. At the time of the sample collection, three viruses, including adenovirus, influenza A, and influenza B, were found mainly to cause URIs among the recruits, and ELISPOT assay for enumeration of the specific antibody‐secreting cells was selected for these viruses. Of the 36 patients with febrile URI studied, viral diagnosis was made in 18 cases, which included 11 patients with adenovirus infection, three with influenza A, and four with influenza B. The first blood sample was collected at the first signs of URI and the second and third samples at 2‐week intervals. The adenovirus‐positive patients developed a strong IgG class antibody‐secreting cell response against the homologous virus, which peaked at the first sample and decreased steeply by the second and third samples. In the influenza A and B patients, the response was similar kinetically to that seen in adenovirus‐positive patients. In those cases where also IgA and IgM class antibody‐secreting cells were determined, the IgG response dominated. The ELISPOT method has potential also as a diagnostic tool for respiratory infections. © Wiley‐Liss, Inc.
Large‐scale screening for human parvovirus B19 DNA in clinical specimens by dot blot hybridization and polymerase chain reaction
Authors: Yoto Yuko , Kudoh Tooru , Haseyama Keiji , Suzuki Nobuhiro , Matsunaga Yasuko , Chiba Shunzo
Published: March 2007

Large‐scale screening for human parvovirus B19 (619) DNA in serum samples was carried out by both dot blot hybridization and the polymerase chain reaction (PCR). Dot blot hybridization was undertaken with a digoxigenin‐labeled DNA probe. Serum samples from four patients were pooled and tested by a dot blot hybridization assay. When a dot was positive, each of the four samples was tested separately to identify the positive sample. The PCR template was the DNA extracted from mixed serum samples from 10 patients. When B19 DNA was positive by PCR, each of the ten samples was tested separately. A total of 7, 969 serum samples were tested by dot blot hybridization and 15 samples (11 patients) were positive for B19 DNA; 7, 038 serum samples were tested by PCR and 71 samples (50 patients) were positive. Large‐scale screening for B19 DNA by PCR suggested a broader spectrum of clinical manifestations associated with B19 infection. © Wiley‐Liss, Inc.
Splice sites of human papillomavirus type 16 E6 gene or heterologous gene required for transformation by E7 and accumulation of E7 RNA
Authors: Belaguli Narasimhaswamy S. , Pater Mary M. , Pater Alan
Published: March 2007

Transformation of primary baby rat kidney cells by the human papillomavirus type 16 (HPV 16) E7 gene and efficient accumulation of E7 RNA have been shown by this laboratory to depend on the integrity of the nucleotide position (nt) 880 splice donor site. Here, the splice sites within the HPV 16 E6 open reading frame (ORF) and the sites of the SV40 splicing unit were examined for an ability to provide this requirement. Constructs containing the HPV 16 E6 sites and the SV40 splice site sequences were used for transformation and RNase protection assays. E6 splice sites supported a low level of transformation, in assays for complete HPV 16 early region constructs containing loss‐of‐function mutations of the nt 880 site. Using constructs with wild‐type E6 or SV40 splice sites showed that both splice sites could substitute similarly for the requirement in cis of the nt 880 site for transformation. HPV 16 E6 mutated splice site and SV40 splice site in reverse, nonfunctional orientation relative to the promoter, were not transformation competent. The HPV 16 E7 RNA levels for the E6 splice site constructs correlated closely with the transformation frequency. The SV40 splice sites were required for E7 transcript accumulation. The results showed E6 splice site function and evidence for enhanced exon skipping from E6 splice donor site to acceptor sites 3′ of the E7 ORF. This was shown with constructs containing loss‐of‐ function mutations of the nt 880 site. These results confirmed the function of the splice sites by the transformation competent constructs and suggested lower transformation frequency than for wild type was due to skipping of the E7 exon. These patterns of transcripts may have a role in the regulation of gene expression during progression to malignancy. The combined results revealed that the general presence of a functional splice donor site was absolutely required for transformation by HPV 16 E7 and accumulation of E7 RNA. © Wiley‐Liss, Inc.
Hepatitis B virus strains in Thailand: Genomic variants in chronic carriers
Authors: Kidd‐Ljunggren Karin , Ekdahl Karl , Öberg Monica , Kurathong Sucha , Lolekha Somsak
Published: March 2007

Genetic heterogeneity of the hepatitis B virus (HBV) has been shown to influence the serological pattern and clinical picture in HBV infection. Thailand has a high transmission rate of HBV, but the molecular epidemiology of HBV strains circulating in this region was hitherto unknown. In this study, the HBV strains from 34 Thai HBsAg‐positive patients were investigated. In a proportion of these samples, an antigenically important region of the S gene (n = 181, and the pre‐S2 and precore genes (n = 15) were sequenced after PCR amplification. Four strains had in‐frame deletions of an upstream region of the pre‐S2 gene, with all deletions ending at the same nucleotide. In one of three anti‐HBe positive strains without a translational stop at codon 28 of the precore gene, there was a one nucleotide insertion in the precore gene. This insertion would cause a frame shift and result in a nonsense protein being expressed, thus providing one explanation for the lack of HBeAg in this patient. Several rare or unique amino acid changes in the region between residues 120 and 161 of the S protein were found. Glycine 145 was changed to alanine in one strain, and this position showed an apparent mixture of glycine and arginine in another. In total, 10 strains displayed unexpected changes that were not related to the normal variability between subtypes or genetic subgroups. It is concluded that there is considerable heterogeneity in HBV strains in Thailand and that this could have clinical and epidemiological importance in a region with high HBV transmission rates. © Wiley‐Liss, Inc.
Molecular epidemiology of an outbreak of hepatitis A in Italy
Authors: Normann Andrea , Pfisterer‐Hunt Mechthild , Schade Stefan , Graff Judith , Chaves Ricardo L. , Crovari Piero , Icardi Giancarlo , Flehmig Bertram
Published: March 2007

The relationship of hepatitis A virus (HAV) isolates associated with an outbreak in Genoa, Italy, in 1993 was examined using direct sequencing of amplicons derived by antigen capture PCR (AC/ PCR) from faecal samples of the infected persons. Forty samples recovered from 38 primary and two secondary cases were examined. The latter were household contacts of the primary cases. In addition, faecal material of 2 unrelated persons infected simultaneously with hepatitis A in Genoa were tested. The PCR products derived rom the P1/P2 junction of the HAV genome were analysed. A 100% nucleotide identity was detected between the viral isolates originating from the primary as well as the secondary cases. The viral isolates recovered from the faecal samples of the two unrelated cases differed from the virus causing the outbreak as well as from each other. These results indicate that a single HAV strain caused the outbreak. The virus might have been transmitted by ingestion of contaminated food or water since all hepatitis A infected employees of the factory had eaten in the same canteen. Definitions of HAV genotypes are based on numerous genetic comparisons of different strains. The sequence comparison of the investigated isolates with published HAV sequences of the P1/P2 genome region revealed that the virus associated with the outbreak belongs to HAV subgenotype IA, whereas the strains recovered from the viral isolates of the unrelated cases belong to subgenotype IB. © Wiley‐Liss, Inc.
Laser coagulation in the upper gi tract: A preliminary light and scanning electron microscopic study
Authors: Meyer H.‐J. , Vonnahme F.‐J. , Huchzermeyer H. , Haverkampf K.
Published: March 2007

Experimentally intact as well as artificially damaged mucosa of porcine stomach was endoscopically coagulated with Nd:YAG laser beams. Energy, intensity pattern, and time of application of the irradiation was varied and supracellular changes of the coagulated mucosa were subsequently investigated with the scanning electron microscope. The mucosa, damaged by laser beams, showed three typical zones: 1. a superficial, central carbonisation zone protected with a thin layer of coagulated mucus; 2. an elevated hyperemic ring with complete loss of superficial epithelium, but intact basement membrane and a blood clotting of the mucosal capillaries; 3. normal mucosa. After coagulating artificially induced bleeding of the mucosa the irradiated centre showed a deep depression with a carbonized surface. Two weeks later regeneration had occurred and there was complete reepithelialisation of the mucosa to about two‐thirds of normal thickness.
Intraocular carbon dioxide laser photosurgery
Authors: Miller James B. , Boyer David S. , Smith Michael R.
Published: March 2007

Intraocular carbon dioxide laser has been used for photocautery and photo‐incision as an adjunct to vitrectomy. This utilizes a new fluid‐tissue interface application for the 10.6 micrometer wavelength infrared radiation. The use of an intraocular probe with an infrared transmitting window through a pars plana incision allows the placement of infrared energy in a fluid medium. The results of animal experimentation and of preliminary clinical trials in human patients are reported. The carbon dioxide laser appears to be a safe and reliable means of intraocular photocautery and photoincision.
Laser applications in urology
Authors: Staehler G. , Halldbrsson T. , Langerholc J. , Bilgram DrIng R.
Published: March 2007

We present the results of experimental and theoretical investigations on the interaction of Nd:YAG laser light with biological tissue in general, and with that of the bladder in particular. A comparison is made of the thermal effects achieved with this laser and those of the argon and CO2 types. A therapeutically safe yet effective dosage for the endoscopic treatment of bladder tumours with the Nd:YAG laser is given. The operating techniques are described along with the first clinical results.
The CO2 laser in the treatment of cavernous haemangioma of the lower lip: A case report
Authors: Ohshiro Toshio
Published: March 2007

A case is reported of a 14‐year‐old girl who had a large cavernous haemangioma and haemangioma simplex of the lower lip. Although a few reports of such a haemangioma and the operation for it can be found in the literature, the blood loss problem and the final cosmesis leave much to be desired. We excised the tumour in two stages using the CO2 laser, retaining full function and obtaining good configuration of the affected part. The suitability of the CO2 laser for this procedure, because of its haemostatic effect, was noted. The importance of both the physiological and psyhcological problems of such a patient were treated by applying our total treatment concept.
Micro‐laser myomectomy technique to enhance reproductive potential: A preliminary report
Authors: McLaughlin David S.
Published: March 2007

Uterine myomas may be removed utilizing the carbon dioxide laser by direct and indirect vaporization, or by direct and indirect excision. The advantages of utilizing the CO2 laser are (1) improved hemostasis; (2) more precision, thus removing only abnormal tissue; and (3) the ability to remove fibroids from previously inaccessible areas. A description of the micro‐surgical technique for myomectomy in eight patients is discussed. No major complications were encountered with this new technique, which will require long‐term follow‐up in a larger series of patients to define more precisely its role in gynecologic surgery.
Observations on the simultaneous use of CO2 and Nd:YAG lasers in neurosurgery
Authors: Fasano Victor Aldo , Benech Franco , Ponzio Roberto Maria
Published: March 2007

Seven cases of cerebral tumors (four deep‐seated sovratentorial gliomas located in motor area, occipital region, parietal region and frontal region; one deep‐seated cystic cerebellar spongioblastoma; one sphenoidal wing meningioma, and one spinal cord intradural tumor) were simultaneously irradiated in the same area with two sources (CO2 and Nd:YAG). Using CO2 and Nd:YAG simultaneously, a larger and deeper lesion was obtained. The main bulk was irradiated with two sources free hand. The implant with the two sources was connected to the operating microscope. The rise in temperature, 3 mm from the border of the lesion, was similar to that obtained with Nd:YAG alone. Hemostasis was not impaired.

In comparison with the single sources, this method allows a more rapid demolition of the tumor without additional damage to the surrounding tissues.
CO2 laser treatment of oral dysplastic precancerous lesions: A preliminary report
Authors: Horch Hans Henning , Gerlach Klaus Louis
Published: March 2007

The oral dysplastic precancerous lesions must be attacked in an early stage, giving the patient a better chance of cure with minimal mutilation. The entire lesion has to be removed in order to prevent local recurrences after treatment. Thirty‐two patients received treatment using the CO2 laser for circumscript disorders of the oral mucosa. The results indicate that CO2 laser treatment represents an alternative technique to the treatment of oral dysplastic precancerous lesions.