bluuebunny/crossref_metadata_2025 huggingface dataset:
Polymerase chain reaction for early detection of HSV‐DNA in cerebrospinal fluid: An experimental mouse encephalitis study
Authors: Boerman Rudolf H. , Arnoldus Edo P. J. , Peters A. C. Boudewijn , Bloem Bastiaan R. , Raap Anton K. , Ploeg Mels Van Der
Published: March 2007

A series of experiments was carried out using a mouse HSV‐1 encephalitis model for detecting HSV‐DNA in CSF by the polymerase chain reaction (PCR). The results were correlated with the time period after corneal inoculation, clinical signs and symptoms, and progression of infection in brain tissue (antigen detection and histo‐pathology), and were compared with virus isolation by spin‐amplified culture (SAC/IF).

The PCR proved to be superior to SACAF, both with respect to early detection and the percentage of positive samples. The results of PCR did not correlate with signs of disease, nor with the interval post‐inoculation. A correlation with the spread of HSV within the brain tissue was found.

The possible pathways for virus spread from brain parenchyma to the CSF as well as a possible explanation for the low amounts of virus and viral‐DNA within the CSF are discussed.
Significance of antibody to hepatitis C virus in Japanese patients with viral hepatitis: Relationship between anti‐HCV antibody and the prognosis of non‐A, non‐B post‐transfusion hepatitis
Authors: Tanaka Eiji , Kiyosawa Kendo , Sodeyama Takeshi , Nakano Yoshiyuki , Yoshizawa Kaname , Hayata Takuro , Shimizu Satoshi , Nakatsuji Yoshiyuki , Koike Yuriko , Furuta Seiichi
Published: March 2007

In a retrospective study, antibody to hepatitis C virus (anti‐HCV antibody) was measured in 80 patients with acute viral hepatitis (type A, 18; type B, 21; type non‐A, non‐B, 41). Anti‐HCV antibody was found in 12 of 20 patients (60%) with non‐A, non‐B post‐transfusion hepatitis (NANB‐PTH) and in 9 of 21 patients (43%) with sporadic NANB hepatitis (NANB‐SPO). Patients with acute hepatitis type A or type B did not have anti‐HCV antibody. The number of patients who developed chronic hepatitis was greater in the group with anti‐HCV antibody than in the anti‐HCV negative group in both NANB‐PTH and NANB‐SPO. The difference was significant in those with NANB‐PTH (P < 0.05).

To investigate the relationship between the long‐term prognosis of NANB‐PTH and the course of anti‐HCV, we studied anti‐HCV antibody in 12 patients who developed chronic type C hepatitis (C‐CH) after PTH and followed them for more than 5 years after the development of PTH. One year after the development of PTH, all 12 had anti‐HCV antibody. Five lost anti‐HCV antibody (group 1) while 7 remained positive (group 2) at the final examination. Four of the 5 patients in group 1 had normal serum transaminases; however, abnormal transaminase persisted in all 7 patients in group 2 until the end of follow‐up (P < 0.05). Because there was no significant difference in the mean follow‐up period between groups 1 (91.6 months) and 2 (102.9 months), it is assumed that the long‐term prognosis of group 1 is better than that of group 2. These observations indicate that anti‐HCV antibody is a useful diagnostic marker for non‐A, non‐B hepatitis in patients with acute hepatitis and that the course of anti‐HCV antibody is closely related to the evolution of serum transaminase abnormality in patients with acute and chronic viral hepatitis type C.
Differential diagnosis of acute viral hepatitis using rapid, fully automated immunoassays
Authors: Eble Kim , Clemens John , Krenc Christine , Rynning Michael , Stojak Joseph , Stuckmann Janice , Hutten Patricia , Nelson Linda , Ducharme Louise , Hojvat Sally , Mimms Larry
Published: March 2007

We report the development of three rapid, fully automated immunoassays allowing the differential diagnosis of acute viral hepatitis. These assays detect HBsAg, IgM antibody to hepatitis B core antigen (IgM anti‐HBc) and IgM antibody to hepatitis A virus (IgM anti‐HAV) using the IMx® instrument system. All IMx assays were run in less than 45 minutes and all steps were fully automated including specimen dilution steps. Specimens from blood donors, diagnostic and hospital patients, and individuals with a variety of infectious and immune diseases were tested for IgM anti‐HAV (n = 1473) or for IgM anti‐HBc (n = 1606) or for HBsAg (n = 9700) by the IMx and commercially available EIA and RIA. Each IMx assay showed 99.8% agreement with current EIA. Reproducibility in all hepatitis IMx assays was significantly better than that observed with manual or semi auto mated assays; within‐run and between‐run % CV ranged from 2.2 to 4.8 and 3.5 to 10.3 respectively. In 29 acute hepatitis B patients studied, HBsAg and IgM anti‐HBc were detected in the first available patient bleed collected from 0 to 4 weeks from the onset of symptoms. IgM anti‐HBc persisted at reactive levels in the IMx assay for 1 to 24 weeks (mean 12.1 ± 5.3 weeks) after the patient presented with symptoms. In individuals exposed to hepatitis A, IgM anti‐HAV was detectable by IMx by 40 days post exposure (average 33.5 days) and IgM had declined to unreactive levels in IMx for all patients by from 3 to 6 months post exposure. These data demonstrate the use of these rapid IMx assays for differentiation of acute hepatitis A and B.
Prevalence of hepatitis C virus antibodies among patients infected with human immunodeficiency virus
Authors: Hayashi Paul H. , Flynn Neil , McCurdy Stephen A. , Kuramoto I. K. , Holland Paul V. , Zeldis Jerome B.
Published: March 2007

A study was undertaken to determine the prevalence and risk factors for serological evidence of hepatitis C virus (HCV) infection in patients infected with the human immunodeficiency virus (HIV). Tests for anti‐HCV antibody were carried out by enzyme‐linked immunoassay (EIA) on 101 HIV‐infected patients from two university‐based outpatient clinics. Anti‐HCV antibody reactive samples were tested by using a recom‐binant immunoblot assay (RIBA) for HCV antibodies. Fourteen of 101 (13.9%) HIV‐infected patients were anti‐HCV reactive by EIA. Of these 14, only seven were reactive by RIBA: four were intravenous drug users as a sole risk factor for HIV infection; and the remaining three acquired HIV by blood transfusion, contaminated instrument exposure or IV drug use and sexual contact. Acquisition of HIV by sexual activity alone was not associated with HCV infection. It is concluded that HCV infection is found in approximately 7% of a university HIV clinic population. False‐positive anti‐HCV antibody serology may lead to overestimation of the prevalence of HCV infection. Female sex and intravenous drug use are significantly associated with HCV infection among HIV‐infected individuals.
Expression and characterization of the preS1 peptide of hepatitis B surface antigen inEscherichia coli
Authors: Lin Yuan , Liu Yan‐Xin , Cislo Teresa , Mason Bobby L. , Yu Mei‐Ying W.
Published: March 2007

The infectious particles of hepatitis B virus (HBV) contain 3 related surface antigens, i.e., small, medium, and large, all of which are encoded by one large open reading frame with multiple initiation codons. The large surface antigen (L‐Ag) contains preS1, preS2, and S regions while both the middle and small surface antigens lack preS1. Several lines of evidence suggested that the preS1 region is involved in the binding of HBV to human hepatocytes as shown by its binding to HepG2 cells and isolated human he‐patocyte membranes. To obtain large quantity of preS1 peptide, an expression vector was constructed containing a lac promoter, the 5′ half of the p‐galactosidase gene, the Factor Xa tetrapep‐tide recognition sequence, and the coding region of preS1 plus preS2. This recombinant plasmid constitutively produced high concentration of a fusion protein in inclusion bodies in Escherichia coli. When the fusion protein was treated with Factor Xa, a peptide consisting of the N‐terminal 91 amino acids of the preS1 region was released. This preS1 fragment purified by anion exchange chromatography was able to bind specifically to the isolated plasma membranes from human liver. Hence, this recombinant preS1 peptide can be used to identify and isolate hepatocyte receptors for HBV.
Characterization of HBV2‐like infections in Spain
Authors: Echevarría José M. , León P. , Domingo C. J. , López J. A. , Echevarría J. E. , Contreras G. , Fuertes A.
Published: March 2007

Hepatitis B viral serum markers suggestive of infection by hepatitis B virus type 2 were found in 354 patients tested in the laboratory during a 2.5 year period of study. Confirmation of the HBsAg reactivity by neutralization and subtyping analysis and determination of serum HBV‐DNA by molecular hybridization were carried out on selected samples from these patients. Clinical and epidemiological data were obtained from 234 patients and serological follow‐up was done in 70 cases. The results obtained from the confirmatory tests and DNA assays indicated that HBsAg reactivities were specific, being frequently associated to low levels of HBV‐DNA. The incidence rates obtained for HBV2 serological patterns among patients at different level of risk for HBV infection, as well as the finding of a high rate of coinfection with hepatitis A virus among those showing acute hepatitis, suggested that the HBV2 agent could be transmitted by the oral route. An hypothesis considering the so called HBV2 infections as resulting from infections by an HBV variant strain which has an improved ability for non‐sexual, non‐parenteral transmission is suggested. Specific recommendations for detection and confirmation of such cases in blood banks are also outlined.
Cytomegalovirus excretion after bone marrow transplantation cannot be predicted from the pre‐transplant level of IgG antibodies
Authors: Pillay D. , Webster A. , Lee D. , Prentice H. G. , Griffiths P. D.
Published: March 2007

The relationship of pre‐bone‐marrow transplant recipient and donor cytomegalovirus (CMV) antibody level to post‐transplant recipient CMV excretion was investigated. CMV excretion was found to be related to pre‐transplant recipient serological status, but not to level of antibody. This therefore contrasts with herpes simplex virus (HSV), for which pre‐transplant antibody level in recipients predicts post‐transplant HSV excretion.
Pre‐S1 and pre‐S2 gene‐encoded proteins in liver and serum in chronic hepatitis delta infection
Authors: Hadziyannis Stephanos J. , Georgopoulou Urania , Psalidaki Eva , Budkowska Agatha
Published: March 2007

Frozen cryostat sections and sera from 30 patients with chronic delta infection were examined for pre‐S1 and pre‐S2 gene‐encoded proteins, and the results were compared to markers in liver and serum HBV and HDV replication. Pre‐S1 and pre‐S2 were detected by indirect immunofluorescence (IF) in the liver in all 26 patients with histochemically demonstrable HBsAg. Pre‐S peptides were found by double IF to have a predominantly cytoplasmic expression and to be located in the same hepatocytes expressing HBsAg. Liver cells expressing hepatitis delta antigen (HDAg) were frequently negative or very weakly positive for HBsAg and pre‐S peptides, but occasional HDAg positive hepatocytes were also strongly positive for HBsAg and for pre‐S peptides, particularly pre‐S2. Circulating pre‐S1 was detected in 24 patients (80%) and pre‐S2 in 27 (90%). Detection of pre‐S peptides in liver and serum was independent of HBV and HDV replication and of the HBV‐DNA integration state. There was no correlation between the amount of circulating pre‐S peptides and serum HBV‐DNA and HDV‐RNA. These results indicate that in chronic HDV infection, formation and secretion of pre‐S peptides and of HBsAg occur independently of HBV and HDV replication and secretion. They further indicate that in the acquisition by replicating HDV of an HBV‐derived envelope in the liver, both HBsAg and pre‐S peptides are concomitantly available but circulating HDV‐RNA is not invariably associated with the presence of these peptides in serum.
Detection of human papillomavirus types 6 and 11 E4 gene products in condylomata acuminatum
Authors: Brown Darron R. , Bryan Janine , Rodriguez Michael , Rose Robert C. , Strike David G.
Published: March 2007

Polyclonal antiserum to an Escherichia coli‐produced beta‐galactosidase/E4 fusion protein of human papillomavirus type 6b (antiserum 256), and affinity purified HPV 11 anti‐E4 antibodies were tested for reactivity in Western blots with bacterially expressed trpE/E4 fusion proteins of HPV types 6b, 11, 16, and 18. To further characterize the affinity purified anti‐E4 antibodies, a dot‐immunobinding assay was performed using overlapping synthetic HPV 11 ETE4 peptides as antigens. Protein extracts of condylomata acuminatum from 18 patients containing HPV type 6 or 11 DNA sequences were tested in Western blots using antiserum 256 or affinity purified HPV 11 anti‐E4 antibodies.

In the Western blots of the trpE proteins, antiserum 256 identified the HPV types 6b and 11 fusion proteins; the affinity purified HPV 11 anti‐E4 antibodies identified only the HPV 11 fusion protein. In the dot‐immunobinding assay, three HPV 11 peptides were recognized, each containing a shared 8 amino acid sequence that differs significantly from the corresponding sequences of HPV types 6b, 16, or 18. In the Western blots of protein extracts from 18 condylomata acuminatum samples shown to contain HPV types 6 or 11 DNA, putative E4 gene products were identified in six samples by antiserum 256. The affinity purified HPV 11 anti‐E4 antibodies identified putative E4 gene products in one of these same six lesions, which was shown to contain HPV 11 sequences by the Southern blot method. All six samples containing E4 gene products were from women. Three of these women were pregnant, one had serum antibodies to the human immunodeficiency virus, and one was a renal transplant recipient receiving glucocorticoids.

This study suggests that unique antigenic sites exist in E4 gene products of HPV types 6 and 11 that distinguish them from other HPV types. Antisera to bacterially expressed E4 fusion proteins can detect putative E4 gene products of HPV types 6 and 11 in condylomata acuminatum from certain patients infected with these HPV types. It may be possible to produce reagents that will detect HPV E4 gene products in a type specific manner in those tissues where these proteins are expressed. These reagents should be useful in determining the conditions leading to E4 gene expression.
Low prevalence of human T‐cell leukemia virus‐I and ‐II infection among drug users in Amsterdam, the Netherlands
Authors: Van Den Hoek J. A. R. , Al E. J. M. , Huisman J. G. , Goudsmit J. , Coutinho R. A.
Published: March 2007

The prevalence of human T‐cell leukemia virus‐I and ‐II infection was studied in a cohort of 346 intravenous and nonintravenous drug users in Amsterdam. Three participants (0.86%) had antibodies to HTLV‐I by two commercially available HTLV‐I enzyme immunoassays (EIA). Infection in these three subjects was confirmed by radioimmunoprecipitation assay. In the immunoblot study, only two of the three subjects were considered positive, since the serum of the third subject had antibodies to p24 only. By means of the polymerase chain reaction two participants (male intravenous drug users infected with human immunodeficiency virus; HIV) appeared to be infected with HTLV‐I and one subject (a male nonintravenous drug user from Surinam) with HTLV‐II. It is concluded that HTLV‐I and HTLV‐II circulate sporadically among drug users in Amsterdam and that risky injecting behaviour, which led to an HIV epidemic among intravenous drug users, has not led so far to an appreciable transmission of the other retroviruses among this group.
Effect of α‐interferon therapy on hepatitis C viraemia in community‐acquired chronic non‐A, non‐B hepatitis: A quantitative polymerase chain reaction study
Authors: Brillanti S. , Garson J. A. , Tuke P. W. , Ring C. , Briggs M. , Masci C. , Miglioli M. , Barbara L. , Tedder R. S.
Published: March 2007

Sera from 30 patients with community‐acquired, biopsy‐proven chronic non‐A, non‐B hepatitis (NANBH) were tested for antibodies to the C100 protein of hepatitis C virus (HCV). The 20 patients who showed reactivity in this assay were followed prospectively for 6 months, during which time seven were treated with recombinant α‐interferon. HCV RNA was detected by “nested” polymerase chain reaction (PCR) in 19 of the 20 anti‐C100‐positive sera taken at the onset of the study and also in five of the ten anti‐Cl00‐negative sera. Pretreatment viraemia levels ranged from 2 × 103 to 2 × 108HCV genomesiml. After 6 months of interferon, elevated serum alanine aminotransferase (ALT) levels had fallen to normal in four of the seven treated patients. In each case the response to interferon was accompanied by either a disappearance of or a decline (1 log to 8 log reduction) in viraemia. HCV genome titres in the three nonresponders and in the 13 untreated anti‐C100‐positive patients did not change significantly over this 6 month period. These findings confirm the aetiological role of HCV in community‐acquired NANBH and suggest that quantitative PCR will become a valuable technique for monitoring the antiviral effect of interferon and other experimental treatments.
Prevalence of HPV cervical infection in a family planning clinic determined by polymerase chain reaction and dot blot hybridisation
Authors: Hallam Nicholas , Green Jonathan , Gibson Patricia , Powis Judith , Bibby Jonathan
Published: March 2007

The overall prevalence of human papillomavirus (HPV) cervical infection in 131 women attending a family planning clinic was 7% (HPV 6/11,16,18,31) by dot blot hybridisation, 53% (HPV 11,16,31) by polymerase chain reaction (PCR), and 56% by the two methods combined. HPV 16 and 18 were the commonest types (4% each) by dot blot, HPV 16 (39%) by PCR. Fifteen percent of subjects had mildly abnormal cervical cytology (grades 1A, 2A, or 3). There was no significant correlation between cytological abnormality and HPV positivity, or between cytological or HPV status and other postulated risk factors for cervical neoplasia.

It is concluded that PCR is considerably more sensitive than dot blot DNA hybridisation in detecting HPV cervical infection in such a “low risk” setting, where HPV copy number may be low. Firm conclusions cannot be drawn from our results regarding a causal role for HPV or other factors in the development of cervical neoplasia.
Molecular epidemiology of a coxsackievirus B3 outbreak
Authors: Rossouw E. , Tsilimigras C. W. A. , Schoub B. D.
Published: March 2007

An outbreak of coxsackievirus B3 infection occurred in South Africa in 1984 with a variety of clinical manifestations being observed. Fifty‐one isolates from patients ranging in age from young babies to middle‐aged adults were obtained. To define further the epidemiology of this outbreak all isolates were characterised by either 1‐ or 2‐dimensional oligonucleotide mapping. One‐dimensional mapping was found to be highly successful for initial screening of the isolates before further characterisation by 2‐dimensional fingerprinting. All isolates were found to be essentially the same strain of coxsackievirus B3 although slight variations in both the 1‐ and 2‐dimensional patterns could be observed. Some coxsackievirus B3 strains from geographically unrelated regions but isolated during the same time period as the outbreak showed clearly distinguishable oligonucleotide maps.
Seroepidemiology of human herpesvirus‐6 in pregnant women from different parts of the world
Authors: Ranger S. , Patillaud S. , Denis F. , Himmich A. , Sangare A. , M'Boup S. , Itoua‐N'Gaporo A. , Prince‐David M. , Chout R. , Cevallos R. , Agut H.
Published: March 2007

The prevalence of HHV‐6 IgG was studied in 11 different countries across several continents: Morocco, Burkina‐Faso, Congo, Ivory Coast, Mali, Niger, Senegal, Togo, Ecuador, Martinique, and France. The study group consisted of 550 pregnant women, representative of the general adult population in each country. Antibodies were detected by immunofluorescence assay on HSB‐2 cells infected with HHV‐6. Each serum was tested at nine dilutions (1:20 to 1:5,120), sera ≥20 being considered positive. For the HSB‐2 cells. Great differences were seen between separate areas: Morocco showed both low prevalence (20%) and a low geometric mean titer (12), whereas sub‐Saharan Africa displayed high prevalences (60% to 90%) and variable geometric mean titers (34 to 229). This study revealed a prevalence of 92% for Ecuador, signifcantly higher than the prevalence for Martinique (50%), yet both countries had very low antibody titers compared with those found in Africa. The prevalence in France (76%) was similar to previous results from other European countries.
Cytomegalovirus infection and excretion in residents of a nursing home
Authors: Stackhouse Frank A. , Kumar Ashir , Maier George A. , Dyke John W.
Published: March 2007

Cy tomegalovirus (CMV), following primary infection, usually remains latent with the potential for reactivation and excretion in saliva and urine. The prevalence of CMV excretion has not been studied among the elderly. This study followed 54 nursing home residents (mean age 83 years and medically stable, but with a wide spectrum of medical conditions). Over a 6 month period these subjects were serially tested for viral excretion in saliva and urine and for CMV antibody. While approximately 93% of the residents tested were sero‐positive, indicating previous CMV infection, there was no evidence of viral excretion in any resident during the study period. Therefore, it is unlikely that the elderly nursing home residents will prove to be a source of CMV to the health care workers or to their family members.
Hepatitis C viraemia in adults with type 2 autoimmune hepatitis
Authors: Garson J. A. , Lenzi M. , Ring C. , Cassani F. , Ballardini G. , Briggs M. , Tedder R. S. , Bianchi F. B.
Published: March 2007

Sera from 14 patients with type 2 autoimmune hepatitis (anti‐LKM, positive) were investigated for evidence of hepatitis C virus (HCV) infection. Antibodies to HCV were detected in 13 patients by both commercial and “in‐house” ELlSAs and also by a second generation recombinant immunoblot assay. Nine of the 13 (69%) anti‐HCV positive patients were shown to be viraemic by a polymerase chain reaction‐based assay for serum HCV RNA. Neither anti‐HCV nor serum HCV RNA were detected in any of 6 controls with primary biliary cirrhosis or in 39 healthy blood donors. These findings strongly suggest a role for HCV in the pathogenesis of type 2 autoimmune hepatitis.
Experimental infection of suckling mice with a host range mutant of junin virus
Authors: Scolaro Luis A. , Mersich Susana E. , Damonte Elsa B.
Published: March 2007

Experimental infection of three mouse strains with a non‐pathogenic mutant of Junin virus named C167 was compared with respect to the parental XJC13 strain. After intracerebral (ic) or intraperitoneal inoculation, XJC13 was highly virulent for 2 day‐old C3H/HeJ, OF1, and BALB/cJ mouse strains, whereas its derivative C167 was attenuated. Survival of the C167‐infected mouse was associated with a restricted replication at the site of inoculation which would impair spread of virus. Thus, the reduced virulence of C167 for suckling mice is independent of the mouse strain and the route of viral entry. When C167 was preinoculated ic I0 days before the challenge inoculation with XJC13 by the same route, mice were partially protected from lethal infection. Since neutralizing antibodies were first detected at 30 days post‐infection, an interference mechanism is postulated as a mechanism of protection of the mice.
Non‐A, non‐B hepatitis in children: A clinical, histologic, and serologic study
Authors: Hsu Shun‐Chien , Chang Mei‐Hwei , Chen Ding‐Shinn , Hsu Hey‐Chi , Lee Chin‐Yun
Published: March 2007

Twenty‐seven children with non‐A, non‐B hepatitis were studied with regard to their clinical, serologic, and histologic features. In contrast to adult patients, non‐A, non‐B hepatitis developed in most pediatric patients (19 out of 27) in the absence of identifiable parenteral exposure, probably because of fewer chances for exposure of children to blood or blood‐contaminated products. Perinatal transmission was considered possible in one of the patients. Fulminant hepatitis had occurred in six patients, two of whom died. It is noteworthy that fulminant hepatitis usually occurred in children with sporadic non‐A, non‐B hepatitis.

The progression to chronic hepatitis after acute illness is less likely in children than in adults. Chronic hepatitis developed in four patients. Two of these were asymptomatic at presentation, suggesting that in some patients, non‐A, non‐B hepatitis infections may remain undetected and progress t o chronic hepatitis. Liver histologic examination was performed in 11 patients. There were no characteristic histologic findings which could differentiate non‐A, non‐B from B hepatitis. Antibodies to hepatitis C virus (anti‐HCV) were tested i n 22 of the 27 patients and 8 (36%) were seropositive for anti‐HCV.
Prevalence of hepatitis C virus infections among heterosexuals with multiple partners
Authors: Van Doornum G. J. J. , Hooykaas C. , Cuypers M. T. , Van Der Linden M. M. D. , Coutinho R. A.
Published: March 2007

A study among heterosexual men and women with multiple sexual partners was carried out to assess the seroprevalence of antibody against hepatitis C virus (HCV). The 468 participants were recruited among visitors to the Clinic for Sexually Transmitted Diseases in Amsterdam. Sera were tested by an enzyme‐linked immunosorbent assay (ELISA; Ortho), a recombinant‐based immunoblot assay (RIBA; Chiron), and the polymerase chain reaction (PCR). A total of 468 persons were tested, and seven (1.5%) were found ELISA positive. Another 25 (5%) were ELISA indeterminate. Six of the seven ELISA‐positive cases were RlBA positive. Further serum samples from five HCV ELISA‐positive persons were tested by PCR, and four were found to be positive. The HCV ELISA‐positive/RIBA‐indeterminate reaction was PCR negative. None of the 17 RIBA‐tested sera of the ELISA‐indeterminate group yielded a positive result. There was a good correlation between an ELISA optical density/ cut‐off ratio greater than 2 and a positive RlBA result. The risk factor for HCV appeared to be the type of sexual partner, i.e., belonging to a “high‐risk” group for human immunodeficiency virus infection and origin from hepatitis B‐endemic countries. It is concluded that HCV may be transmitted through heterosexual contact but probably with low efficiency.
Analysis of a pregnancy‐screening and neonatal‐immunization program for hepatitis B in Hamilton, Ontario, Canada, 1977‐1988
Authors: Chernesky Max A. , Blajchman Morris A. , Castriciano Santina , Basbaum Judy , Spiak Christina , Mahony James B.
Published: March 2007

During the 12 years from January, 1977, to December, 1988, the Hamilton Centre of the Canadian Red Cross Society (CRCS) Blood Transfusion Service screened 98,712 pregnant patients for hepatitis B surface antigen (HBsAg) and identified 120 positives (0.12%). The number of positives ranged from six to 16 per year. We were able to trace and enroll 65 mothers (54%) and 96 of their children in the follow‐up study. The majority of the women were between 20 and 30 years of age (95.4%) and married (86%), and about one‐half were employed outside the home. Sixty‐five percent were white and 34% Asian, and 20 countries were listed as their places of origin. Hepatitis B immune globulin (HBIG) was available for neonatal immunization since 1977 and combined with vaccine since 1982. Of the 96 candidates for HBIG, 60 (63%) received HBIG within 24 hr, one after 3 months, four unknown, and 31 did not receive it. Of the 56 candidates for vaccination from 1982 to 1989,26 (46%) received three doses, seven had two doses, eight had one dose, one was unknown, and 14 had none. HBsAg tests were performed on 69 children (71.8%) and anti‐HBs on 61 (63.5%). Four of the children are HBsAg positive, 31 have anti‐HBs, and 31 have no detectable antibodies. All four HBsAg positives had not received vaccine, and only one had received HBIG. Of the children positive for hepatitis B surface antibodies, five had received no immunization and therefore had been subclinically infected. Of the 32 vaccinees tested for anti‐HBs at least 2 years after immunization was complete, 27 (84.4%) had detectable antibodies. Thus, although several children did not receive appropriate immunization early in the program, the majority did receive both HBIG and vaccine, apparently preventing infection. The consequences of no immunization in these children were a 33.3% infection rate (7/21) and a 14% (3121 ) rate of chronic infection.
Quantitation of HIV: Correlation with clinical, virological, and immunological status
Authors: O'Shea Siobhan , Rostron Timothy , Hamblin Anne S. , Palmer Sara J. , Banatvala Jangu E.
Published: March 2007

A quantitative assay has been used to measure titres of infectious HIV in peripheral blood of symptomatic and asymptomatic patients. Viral titres were assessed in conjunction with virological and immunological status of patients including measurement of p24 antigen, antibody responses to structural (gp41, p24) and regulatory gene products (NEF, REV, TAT, and VIF), determination of beta 2 microglobulin levels and enumeration of lymphocyte subsets. Titres of HIV were significantly higher among symptomatic than asymptomatic patients. Viral load was closely associated with the number of CD4+cells, the proportion of these cells harbouring HIV increasing with disease progression. Higher titres of infectious HIV among symptomatic patients was also associated with p24 antigenaemia and decreased antibody responses to NEF.
Persistence of cytomegalovirus in human long‐term bone marrow culture: Relationship to hemopoiesis
Authors: Preiksaitis Jutta K. , Janowska‐Wieczorek Anna
Published: March 2007

Using pre‐established human long‐term marrow culture (LTMC), we studied cytomegalovirus (CMV) replication in this system after in vitro infection of nonadherent cells obtained from these cultures with CMV AD‐169. After infection with 5 immediate‐early antigen foci/cell CMV was detectable for 63–123 days (peak titer 2.0 × 103−1.3 × 109) in the supernatants of LTMC. Lower MOI resulted in a delay in the detection and longer persistence of CMV in LTMC although peak titers were unchanged. CMV infection was associated with destruction of the stromal layer, appearance of a subset of large (23 μ) CMV‐infected mononuclear cells in the nonadherent fraction, and early differentiation of nonadherent cells into a homogenous population of macrophage‐like cells. CMV infection resulted in a reduction and premature disappearance of committed progenitors (BFU‐E, CFU‐GM) in LTMC. Persistence of CMV in LTMC was linked to ongoing hemopoiesis. Human bone marrow may be an important site for CMV replication during acute infection and CMV persistence.
Detection of B19 Parvovirus in Human Fetal Tissues by Electron Microscopy
Authors: Field Anne M. , Cohen B. J. , Brown K. E. , Mori J. , Clewley J. P. , Nascimento J. P. , Hallam N. F.
Published: March 2007

We present the electron microscopy observations on samples from 38 pregnancies that were investigated for B19 parvovirus infection. Thirty‐four had resulted in fetal loss thought to be due to a virus infection and 22 of the 38 were positive for B19 parvovirus in one or more of the tissues. Twenty‐one placentas and 75 fetal tissue samples were examined. Fresh samples were investigated by immune electron microscopy while formalin‐fixed tissues were examined as thin sections and by negative staining of tissue extracts with direct electron microscopy. Electron microscopy was more sensitive on fresh than on fixed samples. The ultrastructural observations on thin sections of fixed tissues yielded new information locating B19 parvovirus particles in both nucleus and cytoplasm of infected fetal cells. The diagnostic results of the range of electron microscopy assays were compared with those of two hybridization methods. The fresh samples yielded comparable results from electron microscopy and hybridisation assays but on formalin‐fixed materials hybridisation was more sensitive.
Significance of isolated hepatitis B core antibodies detected by enzyme immunoassay in a high risk population
Authors: Kroes A. C. M. , Quint W. G. V. , Heijtink R. A.
Published: March 2007

Serum samples, which were found positive for anti‐HBc and negative for HBsAg and anti‐HBs during routine testing with Abbott enzyme immunoassays (EIA), were collected prospectively. The samples were obtained from patients with a high risk of hepatitis B. Further analysis was carried out using radioimmunoassays and tests for additional markers of hepatitis B. 84 of the 203 initially collected samples (41.4%) were found positive for anti‐HBs by RIA. Of the 119 samples negative for anti‐HBs by RIA, 103 were available for further investigation, and 35 (34.0%) of these were negative for anti‐HBc by RIA. This indicates that low sensitivity of the anti‐HBs EIA and non‐specificity of the anti‐HBc EIA may account for about 60% of the cases presenting with isolated anti‐HBc at initial testing. Fifty‐two samples positive for anti‐HBc by RIA, with isolated anti‐HBc confirmed after RIA testing for HBsAg and anti‐HBs, were examined further: anti‐HBe was detected in 16 sera, anti‐ HDV in one serum and HBV‐DNA in 5 sera by PCR. Additional testing of serum samples with isolated anti‐HBc is warranted.
Acute hepatitis B viral infection in a patient with anti‐HBs
Authors: Wang Yuan‐Jen , Lee Shou‐Dong , Wu Jaw‐Ching , Lo Kwang‐Juei
Published: March 2007

We report a patient with antibody to hepatitis B surface antigen (anti‐HBs) but no antibodies to other hepatitis B virus components, who developed acute symptomatic type B hepatitis. The possible explanations for this unusual serological pattern are 1) the antibody‐positive status, which developed against only a subdeterminant of hepatitis B surface antigen (HBsAg), arose naturally or as the result of cross‐reaction with a variety of antigens; and 2) seroconversion to anti‐HBs occurred in response to surface antigen of a mutant strain of hepatitis B virus (HBV). This anti‐HBs positivity, in the absence of antibody to hepatitis B core antigen, does not provide natural immunization against HBV infection, and so is not protective. Individuals who are positive to anti‐HBs antibody alone which is not elicited by HBV vaccine, should be vaccinated against possible HBV infection.
Recognition of respiratory syncytial (RS) virus proteins by human and BALB/C CD4+ lymphocytes
Authors: Anderson J. J. , Harrop J. A. , Peers H. , Turnbull T. , Toms G. L. , Scott R.
Published: March 2007

Lymphocyte proliferation assays were used to determine the ability of human and BALB/c T‐ lymphocytes to recognise and respond to in vitro challenge with purified preparations of four res piratory syncytial (RS) virus proteins. Human peripheral blood lymphocytes (PBLs) collected from adult donors as well as primed BALB/c mouse splenocytes each responded specifically to challenge with intact RS virus and preparations of the fusion (F), attachment (G),23 kilodalton (23K), and 34K phospho‐ (P) proteins of the virus. F protein was recognised most frequently by human PBLs, and elicited higher levels of response than equivalent concentrations of the other protein preparations examined. The hu‐ man PBL proliferative responses elicited by in vitro challenge with intact virus antigen as well as with each of the four protein preparations were found to be confined to the CD4+ T‐helper (Th) sub‐population of lymphocytes. However, proliferative responses to intact virus and F protein were found to be accompanied by only modest and inconsistent production of Interleu‐ kin‐2 (IL‐2). Finally, no evidence was obtained to indicate that any of the challenge antigens employed in this study were intrinsically mitogenic, as neither naive human cord blood lymphocytes, nor un‐primed BALB/c mouse splenocytes pro liferated when challenged with intact RS virus or with F, G, 23K, or P protein preparations.
Prevention of perinatal transmission of hepatitis B Virus (HB V): A comparison of two prophylactic schedules
Authors: Wheeley Susan M. , Jackson Paul T. , Boxall Elizabeth H , Tarlow Michael J. , Gatrad A. Rashid , Anderson Janet , Bissenden Jeffrey
Published: March 2007

Perinatal transmission of hepatitis B virus (HBV) from HBsAg carrier mothers who were HBeAg+, antiHBe+, or negative for both HBe markers, was interrupted using either 4 doses of vaccine, or one dose of hepatitis B immunoglobulin (HBIG) at birth, combined with 4 doses of vaccine. In those infants who received HBlG at birth, the antiHBs titre was significantly higher at 1 and 2 months old, but at 6, 9, and 18 months old, there was no significant difference.

Among the infants of carrier mothers who did not display HBeAg (i.e., were antiHBe+, or negative for both HBe markers), a transient subclinical infection would have been expected in around 10% had there been no intervention. No evidence of such infection was detected, and no difference in outcome was found between the two treatment groups.

Amongst infants born to HBeAg+ carrier mothers, infection occurred in 1 out of 8 who had received HBlG and vaccine, and in 3 of 8 who had received vaccine only. The difference in outcome was not statistically significant, but the numbers analysed were small. The infections which occurred in spite of prophylaxis may be attributable to in utero infection, poor response to vaccine by the infant, or to the mother having a particularly high HBV‐DNA level. HBlG given at birth to infants of HBeAg+ carrier mothers may enhance the protection of infants who are destined to be poor responders to vaccine.
Hepatitis C virus RNA in saliva of patients with posttransfusion hepatitis and low efficiency of transmission among spouses
Authors: Wang Jin‐Town , Wang Teh‐Hong , Sheu Jin‐Chuan , Lin Jaw‐Town , Chen Ding‐Shinn
Published: March 2007

In a prospective study of posttransfusion hepatitis, 14 patients who were diagnosed with post‐transfusion hepatitis C were enrolled randomly for the study of hepatitis C virus (HCV) RNA in saliva. Saliva and serum samples were collected on the same day. Spouses of 11 married patients were also tested for anti‐C100 and HCV RNA. Paired serum and saliva samples were tested for HCV RNA by a nested polymerase chain reaction (PCR). Two primer pairs specific for the non‐coding region of HCV were used for the PCR and a oligonucleotide sequence between the primers was used as the probe for Southern hybridization. Six patients were positive for HCV RNA by first round PCR amplification and an additional four patients were detected after second round PCR. All patients were negative for HCV RNA in saliva after first round PCR, while seven were positive after second round PCR amplification. All seven patients were positive for HCV RNA in paired serum samples. HCV RNA was detectable in saliva from 1 week to 38 months after the onset of hepatitis. All spouses were negative for anti‐100 and HCV RNA. We conclude that HCV RNA is present in the saliva of approximately half of patients with acute and chronic hepatitis C, and the presence of HCV RNA correlates with HCV viremia. The efficiency of HCV transmission is low among spouses.
Correlation between cyclophosphamide‐induced viral susceptibility and depletion of junin virus—induced suppressor populations
Authors: Campetella Oscar E. , Galassi Nora V. , Barrios Hebe A.
Published: March 2007

In contrast to lymphocytic choriomeningitis virus, another arenavirus, Junin virus (JV), the etiologic agent of Argentine hemorrhagic fever, when inoculated into suckling mice, induces lethal meningoencephalitis characterized by a delayed‐type hypersensitivity (DTH)‐like immune response. However, the adult BALB/c mouse is resistant to infection and no DTH reaction can be seen. This different viral sensitivity may be related t o the development of an antigen non‐specific DTH‐suppressor cell pathway at work in the adult mouse. When the resistant mice are treated with cyclophosphamide (Cy) (50 mg/kg each dose) given at days −1,+1,+4 (zero: infection day), animals become susceptible and develop DTH reaction in brain that leads to death. We analyze the influence of the timing of Cy administration on the suppressor system developing after infection. It was found that Cy depletes the previously described JV‐induced suppressor populations (Tsv) but a new suppressor cell (Tsv*) is disclosed bearing the Thy 1 + Ly1+2− phenotype which is unable to depress DTH in Cy‐treated animals. With only two doses of Cy corresponding to days −1 and + 1, the target of Tsv* cells is depleted but the third dose is still required to achieve full depletion of Tsv cells which are able to employ the Cy‐resistant antigen‐specific suppressor cells as targets. Since the Cy treatment is able t o deplete the Tsv population together with the target of Tsv* cells, animals became unable to regulate lethal DTH reaction. Thus, a cellular explanation for an empirically established Cy schedule able to abrogate the adult mouse resistance to JV is proposed.
Detection of HPV DNA in archival specimens of cervical cancer using in situ hybridisation and the polymerase chain reaction
Authors: Thompson Carol H. , Rose Barbara R. , Cossart Yvonne E.
Published: March 2007

An archival survey of 98 cervical cancer specimens dating from the 1920s to the 1980s was undertaken to determine whether changes had occurred in the prevalence of human papillomavirus (HPV) DNA. HPV DNA was detected in paraffin sections of cancers fixed in 10% formalin by in situ hybridisation (ISH) using HPV 6, 11, 16, and 18 32P‐labelled DNA probes under conditions of high stringency; and by the polymerase chain reaction (PCR) using 20‐mer oligonucleotide primers to amplify 109 bases of the E6 region of HPV 16. In 30 instances results obtained from Southern blot hybridisations which had been carried out on specimens of fresh tissue from the same cancers collected during the 1980s were available for comparison. The rates of HPV DNA detection in cervical cancers ranged from 83% (by Southern or PCR) and 70% (by ISH) on specimens from the 1980s, to 50% and 63% (by ISH and PCR, respectively) on specimens from the 1920s. HPV 16 was by far the most common type, being identified by Southern or ISH i n approximately 92% of HPV DNA‐ positive specimens. No significant change in the prevalence of HPV DNA, or of HPV types, in cervical cancers was found over the 65 year period examined.
Immunoglobulin G subclasses and lymphocyte stimulatory responses to cytomegalovirus in transplant patients with primary cytomegalovirus infections
Authors: Mathiesen T. , Brattström C. , Andersson J. , Linde A. , Ljungman P. , Wahren B.
Published: March 2007

The early activation of T‐ and B‐cell responses to cytomegalovirus was studied in immunosuppressed patients. Primary lymphocyte stimulation to cytomegalovirus (CMV) antigen, a measure of T‐helper activity, and anti‐CMV IgG subclass responses were analyzed. Ten patients suffering from primary CMV infection following renal transplantation were studied. Of the ten, nine became positive for CMV induced lymphocyte proliferation 5‐40 weeks after transplantation. Nine showed an almost simultaneous appearance of anti‐CMV IgG1 and three at 3‐32 weeks after transplantation, while one patient synthesized only low levels of anti‐CMV IgG1. The lymphocyte proliferation assays have limited diagnostic value for primary CMV infection in renal transplant patients. The humoral and cellular immune responses seemed to be independent of each other.
Low risk of transmission of the human immunodeficiency virus by a solvent‐detergent‐treated commercial factor VIII concentrate
Authors: Paolantonio Tiziana D. , Mariani Guglielmo , Ghirardini Alessandro , Gringeri Alessandro , Mannucci Pier Mannuccio , Mastrullo Lucia , de Biasi Raffaello , Giustarini Gloria , Morfini Massimo , Schiavoni Mario , Ciavarella Nicola , Zehender Gianguglielmo , Zanetti Alessandro Remo
Published: March 2007

A study evaluating the risk of a commercial factor VIII (FVIII) concentrate's transmitting the human immunodeficiency virus (HIV) was carried out on hemophiliacs, by using multiple serological markers and the polymerase chain reaction (PCR). Twenty‐nine hemophiliacs, negative for HIV antibodies, were treated for 18 months with a concentrate that had been inactivated by solvent‐detergent. HIV‐1 antibodies and antigen were assayed during the follow‐up period. At the end of the study, all patients were also tested by the HIV 1+2 combined antibody assay; Western blot (WB) antibody analysis; and in eight cases, by an HIV‐1 PCR technique. Patients received a yearly median FVIII dose of 35,330 IU (range 3,300‐306,000); the median number of lots given to each patient was 6 (1‐45). During the follow‐up period and at the end of the study, HIV‐1 antibodies and antigen were not detected in any of the subjects. The HIV 1+2 combined assay and WB analysis carried out only at the end of the study were negative. HIV‐1 PCR was negative in all the tested patients. This study has shown that this solvent‐detergent‐treated FVIII concentrate did not transmit HIV.
Association of hepatitis E virus with an outbreak of hepatitis in Pakistan: Serologic responses and pattern of virus excretion
Authors: Ticehurst John , Popkin Terry J. , Bryan Joe P. , Innis Bruce L. , Duncan J. Fred , Ahmed Aftab , Iqbal Muhammad , Malik Iftikhar , Kapikian Albert Z. , Legters Llewellyn J. , Purcell Robert H.
Published: March 2007

Hepatitis E virus (HEV), a positive‐strand RNA agent, has been associated with enterically transmitted non‐A, non‐B hepatitis in Asia, Africa, and Mexico. To evaluate the role of HEV in an outbreak of hepatitis in Pakistan, we used immune electron microscopy to detect 1) antibody to HEV, for evidence of infection, and 2) virus, to determine the pattern of HEV excretion. Paired sera from 2 patients were assayed for antibody by using reference HEV: one seroconverted, an atypical finding for HEV infections; the other had high levels of anti‐HEV in both sera. Virus particles with the size (29 × 31 nm) and morphology of HEV were detected in feces from 10 of 85 patients and serologically identified as HEV by using reference antibodies from an HEV‐infected chimpanzee. One of these HEV‐containing specimens was collected 9 days before the onset of jaundice; it was among feces from 38 outpatients with nonspecific symptoms and biochemical hepatitis, 12 of whom subsequently developed jaundice. The other 9 feces with HEV were among 36 collected within 7 days of the onset of acute icteric hepatitis; all 11 feces from days 8 to 15 were negative for HEV. Fecal concentrations of HEV appeared t o be lower than those of many enteric viruses: only one specimen contained as many as 5 particles per EM grid square. It is concluded that HEV was etiologically associated with the epidemic and was predominantly excreted at very low levels during the first week of jaundice.