bluuebunny/crossref_metadata_2025 huggingface dataset:
Cocaine injection and ethnicity in parenteral drug users during the early years of the human immunodeficiency virus (hiv) epidemic in new york city
Authors: Novick David M. , Trigg Harold L. , Des Jarlais Don C. , Friedman Samuel R. , Vlahov David , Kreek Mary Jeanne
Published: March 2007

Parenteral drug users have a high prevalence of infection with human immunodeficiency virus (HIV), the etiologic agent of acquired immune deficiency syndrome (AIDS). New York City has had a prolonged and extensive epidemic of HIV infection and AIDS. In this study, we analyze, in relation to antibody to HIV (anti‐HIV), available data from sera from parenteral drug users collected in New York City during 1978 through 1983 in the course of studies of liver disease. Among parenteral users of both heroin and cocaine, 30 (52%) of 58 had anti‐HIV, compared with six (13%) of 48 injectors of heroin only (P < 0.0001). Only two (11%) of 18 white patients were HIV‐infected, compared with 34 (39%) of 88 black or Hispanic patients (P = 0.03). No other factors studied were linked to anti‐HIV. In a multiple logistic regression, anti‐HIV was significantly more common in parenteral users of both cocaine and heroin (P < 0.0001), black patients (P = 0.02), and Hispanic patients (P = 0.049). We conclude that parenteral users of both cocaine and heroin as well as black and Hispanic patients were disproportionately HIV‐infected during the early years of the HIV epidemic. Use of cocaine and heroin as well as ethnicity were independently linked to anti‐HIV. Measures to prevent or treat drug use, HIV infection, and other medical problems while addressing the specific needs of cocaine users and black and Hispanic patients are urgently needed.
Detection by antibody probes of human papillomavirus type 6 E5 proteins in respiratory papillomata
Authors: Chen Show‐Li , Mounts Phoebe
Published: March 2007

We have demonstrated the expression of proteins arising from the E5a and E5b open reading frames (ORFs) of human papillomavirus type 6c (HPV‐6c) in respiratory tract papillomata. Recombinant plasmids were constructed to express the ORFs in the bacterial vectors pATH and pRIT2T. Fusion proteins were purified and injected into rabbits to produce polyclonal antibodies. Characterized antibodies generated against these fusion proteins were used in immunoperoxidase assays to identify the presence and distribution of HPV‐6 E5 proteins in biopsy specimens of respiratory tract papillomata. The results showed that the E5a and E5b proteins were distributed throughout the thickness of the epithelium in the papillomata but not in the basal layer. The proteins were found in nuclei and in the cytoplasm of koilocytotic cells. Positive reactivity with a similar distribution in the epithelium and subcellular location was obtained in papillomata induced by other HPV‐6 subtypes. This cross‐reactivity was not unexpected, since nucleotide and amino acid (aa) sequence comparisons between HPV‐6c and ‐6e demonstrated 79% sequence identity with 15 aa substitutions in the 91 aa of E5a. The E5b ORF of HPV‐6c has the potential to encode a protein of 74 aa that differed at 28 positions compared with the 72 aa of HPV‐6e.
Cytomegalovirus cultures during maintenance DHPG therapy for cytomegalovirus (CMV) retinitis in acquired immunodeficiency syndrome (AIDS)
Authors: Jennens Ian D. , Lucas Charles R. , Murray Sandland A. , Maclean Hector , Hayes Kathleen
Published: March 2007

Nine patients with acquired immunodeficiency syndrome (AIDS) and cytomegalovirus (CMV) retinitis on maintenance therapy with ganciclovir: 9(1,3‐dihydroxy‐2‐propoxymethyl) guanine (DHPG) at high dose (30 mg/kg/week) or low dose (20 mg/kg/week) were tested every 1–2 weeks for CMV isolation from blood, saliva, and urine. Duration of therapy ranged from 1.5 to 12 months (average 5.3 months). During pretreatment and low‐dose and high‐dose maintenance therapy, CMV was isolated from 48/59 (81%), 90/211 (43%), and 40/290 (14%) of specimens, respectively. Three patients with progressive retinitis had viraemia more frequently than did six patients with stable retinitis, CMV being isolated from 29/47 (62%) and 17/121 (14%) of blood samples, respectively.
Three serotypes of norwalk‐like virus demonstrated by solid‐phase immune electron microscopy
Authors: Lewis David C.
Published: March 2007

Solid phase immune electron microscopy (SPIEM) was used to investigate the serological differences between Norwalk‐like virus (NLV) strains from five different outbreaks within the United Kingdom.

The existence of two previously demonstrated serotypes, Lewis et al. (Journal of Clinical Microbiology 26:938–942, 1988), was confirmed by the use of whole convalescent sera and purified IgM. A third serotype was found to be the agent of two recent hospital outbreaks and could similarly be typed by use of whole sera or pure IgM. Paired sera were available for two of the three serotypes and demonstrated rising antibody levels. These antibody rises were also specific for the infecting serotype. However, two serum pairs from a later outbreak gave antibody rises to all three serotypes, although much higher counts were produced with the infecting serotype.

SPIEM is a useful method for distinguishing NLV serotypes and can also be used to detect specific IgM and to demonstrate seroconversion. Cross‐reacting antibodies, possibly anamnestic in origin, can occur after natural infection and could cause confusion in typing virus unless further evidence of the identity of the infecting agent is obtained.
Frequent shedding of human herpesvirus 6 in saliva
Authors: Harnett G. B. , Farr T. J. , Pietroboni G. R. , Bucens M. R.
Published: March 2007

We have previously reported the isolation of HHV‐6 from saliva samples. Because these isolations were made in PHA‐stimulated lymphocytes from healthy adults, which may occasionally contain endogenous HHV‐6, it was desirable to repeat this work using cord blood lymphocytes. In this study 18 isolations of viruses provisionally characterized as HHV‐6 were made from 19 saliva samples by centrifugally enhanced inoculation into PHA‐stimulated cord blood lymphocytes. HHV‐6 was not found in 10 pernasal aspirates, 50 endocervical swabs, or 30 male urethral swabs. It is concluded that HHV‐6 is usually present in the saliva of most adults and that this affords a possible explanation of the high infection rate with this virus in young children.
HIV antibodies in whole saliva detected by ELISA and western blot assays
Authors: Holmström P. , Syrjänen S. , Laine P. , Valle S‐L. , Suni J.
Published: March 2007

Paired serum and saliva samples were tested by enzyme‐linked immunosorbent assay (ELISA) and Western blot (WB) for the presence of human immunodeficiency virus (HIV) antibodies. The study group included 36 individuals known to be HIV seropositive and 14 healthy, seronegative controls. HIV antibodies were detected in all but one of the saliva samples of the seropositive subjects. In this particular patient, seroconversion was documented 1 week earlier by sequential testings. A further saliva sample obtained 2 months later was ELISA positive for salivary HIV antibodies.

Antibodies against HIV proteins gp 120 and gp 160 were detected by Western blot assay in all saliva specimens taken from HIV seropositive subjects (including the ELISA‐negative patient who seroconverted. Antibodies against other viral proteins (p65, p51, gp41, p35, p24 p18) were found in saliva haphazardly without any clear‐cut correlation with the clinical stage of the disease. Pretreatment of the saliva with protease inhibitor was essential for the diagnostic use of saliva for the detection of HIV antibodies by Western blot assay.

Calculation of the ratio of titres in serum to those in saliva showed the highest ratios in symptomless subjects (mean ± SD; 1844 ± 1412) and the lowest in patients with acquired immune deficiency syndrome (AIDS) (mean ± SD; 811 ± 445). The ratio of serum to saliva by ELISA showed a positive correlation with salivary flow rate, indicating a dilution of salivary HIV antibodies with increasing salivary flow rate. The gingival bleeding index was negatively correlated with the ratio, supporting the concept that salivary HIV antibodies transudate from blood to saliva via gingival fluid.
Overview of biological effects of addition of DNA molecules to cells
Authors: Temin Howard M.
Published: March 2007

Injected DNA proceeds with certain probabilities through the following steps: degradation by serum nucleases, adsorption to cells, uptake into cells, ligation to other DNA, mutation, expression of unintegrated DNA, integration, expression of integrated DNA, and activation of or inactivation of cellular genes. The maximal probability per DNA molecule of each of these steps is estimated based on experimental results in cell culture with transfection of DNA and with infection by retroviruses. A maximum cumulative probability of having a harmful effect is calculated to be less than 10−16 to 10−19 per DNA molecule from a cell without activated proto‐oncogenes or active viral oncogenes. The most frequent harmful effects considered are inactivation of a tumor suppressor gene and activation of a proto‐oncogene. Such inactivation and activation in a cell that could give rise to cancer would increase the age‐standardized incidence of cancer by a small amount. The amount of increase would differ among individuals depending upon their genotypes and their environments. Thus, the magnitude of the increase will depend upon the frequency of more sensitive individuals. The probability of an increased incidence of cancer as a possible effect of the vaccination should be compared to the number of DNA molecules to be injected per person and to the protective effects of a successful HBV vaccine.
High levels of Epstein‐Barr virus in the oropharynx: A predictor of disease progression in human immunodeficiency virus infection
Authors: Diaz‐Mitoma Francisco , Ruiz Alejandra , Flowerdew Gordon , Houston Stanley , Romanowski Barbara , Kovithavongs T. , Preiksaitis Jutta , Tyrrell D. Lorne
Published: March 2007

The role of Epstein‐Barr virus (EBV) on the progression of human immunodeficiency virus (HIV) infection is not well defined. The objective of this prospective study was to determine the prevalence of EBV excretion and the rol that EBV might have on HIV disease progression. Fifty‐two homosexual males were studied, all of whom had positive EBV serology. Twenty‐four of the 27 HIV‐seropositive and 14 of the 25 HIV‐seronegative subjects had detectable levels of EBV DNA in oropharyngeal cells. In addition to a greater prevalence of detectable EBV, the level of excretion was higher among HIV‐seropositives than among HIV‐seronegatives, and higher among group III than among group II HIV‐seropositive men. These results are consistent with earlier studies showing a relationship between immunosuppression and EBV reactivation. The EBV excretion levels in a control group of 52 age‐matched heterosexual males were substantially lower than those found in the homosexual group.

In a proportional hazards regression analysis EBV excretion was found to be the best single predictor of progression of HIV infection (P<0.001). HIV p24 core antigenemia (P=0.048) and low EBNA (P=0.024) were significant predictors independent of EBV excretion. Whether EV directly accelerates the time to progression or is merely a marker of underlying subclinical immunosuppression remains an open question.
Antiviral strategies in chronic hepatitis B virus infection: I. Establishment of an in vitro system using the duck hepatitis B virus model
Authors: Bishop Naomi , Civitico Gilda , Wang Yanyan , Guo Kejian , Birch Chris , Gust Ian , Locarnini Stephen
Published: March 2007

Primary duck hepatocyte (PDH) cultures were established from ducklings congenitally infected with the duck hepatitis B virus (DHBV), plated onto feeder cell layers of irradiated human embryonic lung fibroblasts, and observed for 2 to 3 weeks. This system permitted the survival of the PDH in a differentiated form free of fibroblastic overgrowth for at least 3 weeks. The hepatocytes were shown to contain all the replicative DNA intermediates found during DHBV replication as well as the DHBV structural proteins PRE‐S1, PRE‐S2, and S of duck hepatitis B surface antigen (DHBsAg). The pool of supercoiled (SC) DHBV DNA increased dramatically from days 10 to 14 postplating. This PDH‐feeder cell layer cell culture model provides a convenient system to study the effects of conventional inhibitors of DHBV replication and compounds targeted at the supercoiled form of DHBV DNA. This approach should allow the evaluation of a variety of strategies for treating chronic carriers of hepadnaviruses.
Comparative studies of HSV‐1 antigens solubilised from infected cells by using non‐ionic or zwitterionic detergents
Authors: Jennings R. , Erturk M.
Published: March 2007

HSV‐1 antigen preparations solubilised from Vero cells by using either the non‐ionic detergent Nonidet P40 or the zwitterionic detergent Empigen BB, and purified on sucrose density gradients or over a sucrose cushion, were tested by ELISA with anti‐HSV‐1 glycoprotein monoclonal antibodies and by radioimmunoprecipitation (RIP) with polyclonal HSV‐1 antiserum. Amongst several proteins detected in these preparations, the four major HSV‐1 glycoproteins, gB, gC, gD, and gE, were found to be present. Differences between NP40 or Empigensolubilised HSV‐1 antigen preparations with respect to two of these glycoproteins, gB and gE, were detected by using a small panel of monoclonal antibodies. Comparative studies in mice showed the Empigen‐solubilised HSV‐1 antigen preparations elicited greater antibody responses and greater protection against lethal HSV‐1 challenge infection than the NP40‐solubilised preparation.
Nosocomial outbreak of neonatal gastroenteritis caused by a new serotype 4, subtype 4B human rotavirus
Authors: Gerna Giuseppe , Forster Johannes , Parea Maurizio , Sarasini Antonella , Di Matteo Angela , Baldanti Fausto , Langosch Beate , Schmidt Sigrid , Battaglia Massimo
Published: March 2007

A nosocomial outbreak of rotavirus gastroenteritis involving 52 newborns occurred between June and September 1988 at the University Children's Hospital of Freiburg, Federal Republic of Germany. Stools from 27 representative patients were examined for rotavirus serotypes, using a monoclonal antibody‐based enzyme‐linked immunosorbent assay. The electropherotype was also examined by polyacrylamide gel electrophoresis of genomic RNA. As many as 18 patients were found to be infected by serotype 4, subtype 4B strain, and in all of them the same electropherotype was detected. Although rotavirus from the remaining nine patients could not be typed, the electropherotype in four was identical to that of the serotype 4, subtype 4B strain. Thus, most of the patients in the outbreak were infected by the same rotavirus strain. Retrospective epidemiological studies showed that the 4B strain began to circulate at the hospital in January 1988, whereas only rotavirus serotypes 1, 3, and 4A were detected in 1985–1987. The primary case of the outbreak was presumably a newborn with acute gastroenteritis, admitted to the hospital from a small maternity unit in the same urban area. During the outbreak, 12 of 44 healthy newborns in the nurseries of the Children's Hospital and other maternity hospitals were found to be asymptomatic rotavirus carriers, and in three of the newborns the same 4B strain was detected. This is the first reported outbreak caused by a serotype 4, subtype 4B strain.
Colposcopy, punch biopsy, in situ DNA hybridization, and the polymerase chain reaction in searching for genital human papillomavirus (HPV) infections in women with normal PAP smears
Authors: Syrjänen Stina , Saastamoinen Jukka , Chang Fuju , Ji Hongxiu , Syrjänen Kari
Published: March 2007

To assess the prevalence of HPV infection in the genital tracts of women with normal PAP smears, a random series of 109 women was reexamined using colposcopy, a further PAP smear, and punch biopsies taken from the cervix (in 33 cases) vagina (212 cases), and anus (20 cases). The biopsy material was examined using routine histological investigations, in situ hybridization (ISH) with a 35S‐labelled DNA probe cocktail (HPV 6,11,16,18), and the polymerase chain reaction (PCR) to detect HPV DNA. Changes consistent with HPV infection were seen in 6.9% (18/262) of the biopsy specimens. Seven biopsy specimens (2.7%) from seven different women were found to contain HPV DNA using ISH. All of these ISH‐positive lesions were diagnosed as morphologically characteristic HPV lesions: six flat condylomas and one papilary condyloma. Using PCR, the HPV DNA detection rate was highest in the cervical biopsy specimens (50%) and lowest (28.6%) in the anal biopsy specimens. A total of 35.5% of the 93 biopsy specimens studied using PCR contained HPV DNA. The commonest type was HPV 11 (54.5%), followed by HPV 18 (33.3%). Four of the nine biopsy specimens (44.4%) from colposcopically normal areas proved HPV DNA‐positive using PCR. Of 17 biopsy specimens in which the histology was normal, seven were examined using PCR and three were DNA‐positive. The discovery of HPV DNA using PCR in 32/92 of the biopsy specimens (34.8%) which had been found to be HPV DNA‐negative when routine ISH was used is noteworthy. The results suggest that the light microscopy criteria currently used in diagnosing HPV infections are of no value in predicting latent HPV infections and that acetowhite staining is unable to distinguish between subclinical and latent infections on the one hand and changes unrelated to HPV on the other.
Study of anti‐dengue NS1 antibody by Western blot
Authors: Kuno G. , Vorndam A. V. , Gubler D. J. , Gómez I.
Published: March 2007

The presence of anti‐nonstructural protein (NS1) antibody in natural dengue infections has been suspected to be associated with development of dengue haemorrhagic fever (DHF). The Western blot technique was used to study the dynamics of the immune response to NS1 and to determine the frequency of anti‐NS1 antibody among confirmed dengue patients in Indonesia, where DHF is common, and in Puerto Rico, where DHF is less frequently observed. Anti‐NS1 antibody was rarely found in those with primary infections in either group. The antibody occurred at a significantly higher frequency in acute‐phase serum samples from secondary infections in Indonesia than in those from Puerto Rico. No difference was observed, however, in Indonesian patients with secondary infection who had dengue fever or DHF.
A precore‐defective mutant of hepatitis B virus associated with e antigen‐negative chronic liver disease
Authors: Ulrich Paul P. , Bhat Ramesh A. , Kelly Irene , Brunetto Maurizia R. , Bonino Ferruccio , Vyas Girish N.
Published: March 2007

The pathogenesis of chronic liver disease (CLD) due to persistent hepatitis B virus (HBV) infection has not been defined, but the disease activity is believed to correlate with the presence of hepatitis B e‐antigen (HBeAg) antigenemia and high viremia. The molecular characterization of an HBV mutant isolated from an HBeAg‐negative patient with severe CLD required amplification of the circulating HBV DNA (2 pg/ml) by the polymerase chain reaction (PCR). Direct sequencing of the nucleotides from five independent amplifications of the conserved precore region consistently revealed a G to A mutation in each of the two terminal codons of the precore region. Codon 28 was mutated from tryptophan‐encoding TGG to a translational stop codon, TAG; codon 29 preceding the core initiation codon was changed from GGC to GAC. For biologic evaluation of these mutations on HBV replication and expression of HBeAg in vitro, HepG2 cells were transfected with cloned, recicularized mutant HBV DNA. The transfected cells contained subviral core particles in the cytoplasm and secreted mature HBV, without HBeAg, into the medium. The findings present the first evidence that complete HBV genomes can be amplified by PCR and are replication‐competent in vitro. The data also indicate that HBeAg is not necessary for replication of HBV and furthermore suggest that HBeAg is not required for the progression of HBV‐induced CLD.
Experimental keratitis in rabbits by human HSV‐1 variants: Prevention and treatment
Authors: Manservigi R. , Incorvaia C. , Di Luca D. , Rotola A. , Balboni P. G. , Sebastiani A. , Rossi A. , Cassai E.
Published: March 2007

The efficacy of different therapies and vaccine preparations was assessed for treating or preventing herpetic ocular keratitis induced by experimental inoculation in rabbits with two HSV‐1 variants that display different pathogenetic potential. Early administration of acyclovir (ACV) promoted fast healing and prevented neurologic involvements: α‐Interferon (α‐IFN) was less efficent than ACV; combined therapy with both drugs increased the antiviral effects. In an attempt to prevent the disease, rabbits were vaccinated with a slightly pathogenic HSV‐1 variant or with a secreted form of an engineered HSV‐1 glycoprotein gB (gB‐1s) and were subsequently challenged with a highly pathogenic HSV‐1 variant. Immunization of rabbits with gB‐1s was much more efficient than immunization with live virus in reducing the severity of herpetic keratitis and in preventing CNS disease.
Predictive value of cytomegalovirus viraemia for the occurrence of CMV organ involvement in AIDS
Authors: Salmon Dominique , Lacassin Flore , Harzic Martine , Leport Catherine , Perronne Christian , Bricaire François , Brun‐Vezinet Françoise , Vilde Jean‐Louis
Published: March 2007

In HIV‐infected patients, the presence of cytomegalovirus (CMV) viraemia is predictive of the development of acquired immunodeficiency syndrome (AIDS), but it is not known whether it predicts further occurrence of CMV organ involvement.

To assess the predictive value of CMV viraemia in the occurrence of CMV organ involvement, CMV blood cultures were performed at the onset of AIDS in 71 patients. They were prospectively followed up for at least 6 months, with a mean of 16.6 ± 7 months (6–36 months).

CMV viraemia was present in 28/71 patients (39.5%) at the onset of AIDS. CMV organ involvement occurred in 18/71 patients (25.4%) after 8.7 ± 3.3 months. Fourteen of the 28 patients (50%) with early CMV viraemia developed CMV organ involvement after 7.7 ± 6.3 months as compared with 4/43 patients (9.3%) who were not viraemic at the onset of AIDS with a mean follow‐up of 11.8 ± 3.8 months (P < 0.001). At the time of CMV organ involvement, CMV viraemia, however, was present in 94.4% of the cases. No difference in survival was observed between initially viraemic and non‐viraemic patients. No difference has found in mean CD4 cell count between viraemic and non‐viraemic patients.

This high predictive value of early CMV viraemia in AIDS for the occurrence of CMV organ involvement underlines the need for repeated search for CMV organ involvement when CMV viraemia is detected, and for less toxic antiviral drugs that might be used earlier.
Sensitive enzyme‐linked immunosorbent assay for antibody to varicella‐zoster virus using purified VZV glycoprotein antigen
Authors: Wasmuth Edward H. , Miller William J.
Published: March 2007

The development of an ELISA of increased sensitivity has permitted a more critical evaluation of human humoral immune responses to the live attenuated varicella (Oka/Merck) vaccine. For use as a solid‐phase antigen, the glycoprotein (gp) antigens are prepared by lectin‐affinity chromatography from lysates of VZV‐infected MRC‐5 cells. The lot‐to‐lot variation in VZV gp content is controlled by standardization of antigen against a panel of human serum providing antigencoated plates of consistent quality. The increased sensitivity of the gpELISA over the VAR ELISA is reflected in the greater seroconversion rate and prepositive rate specificity. These determinations have been shown to be specific for anti‐VZV by absorption experiments using purified VZV gp antigens.
Prevalence of human parvovirus (B19) and rubellavirus infections in urban and remote rural areas in northern Brazil
Authors: de Freitas Ronaldo B. , Wong Derek , Boswell Fiona , de Miranda Mario F. R. , Linhares Alexandre C. , Shirley Jane , Desselberger Ulrich
Published: March 2007

Sera from inhabitants of Belém, Pará (542 sera), Brazil and of members of 3 Brazilian tribes—Tiriyo/Alto Paru (near Surinam) (212 sera), Xicrin (128 sera), and Mekranoiti (121 sera)—of different age and sex groups were tested for the presence of specific antibody against human parvovirus (B19) (RIA) and rubellavirus (latex agglutination test).

Parvovirus (B19) IgG was found in 42.6% of the population sample from Belém but in only 4.7 to 10.7% of the members of the tribes. Rubellavirus antibody was found in 72.7% of the sera from Belém but approaching a prevalence of 85–90% in age groups above 20 years. In the tribes rubellavirus antibody was detected in 36.9 to 72.6% of all sera. There were remarkable sex differences of antibody prevalence in several age groups of the population from Belém and of the tribal populations.

About a quarter of the skin rashes in Belém that were not attributable to infections with rubella, measles, or arboviruses were caused by recent B19 infections.
In situ hybridization detection of HTLV‐I RNA in peripheral blood mononuclear cells of TSP/HAM patients and their spouses
Authors: Beilke M. A. , In D. Riding , Gravell M. , Hamilton R. S. , Mora C. A. , Leon‐Monzon M. , Rodgers‐Johnson P. E. B. , Gajdusek B. C. , Gibbs C. J. , Zaninovic V.
Published: March 2007

This is the first report of the direct detection of HTLV‐I RNA in uncultured peripheral blood mononuclear cells (PBMNC's) of patients with tropical spastic paraparesis and HTLV‐l‐associated myelopathy (TSP/HAM) and their spouses, using the technique of in situ hybridization. Twenty‐one Colombian patients were tested, all of whom had antibodies to HTLV‐I; the presence of HTLV‐I proviral DNA in their PBMNC's was confirmed by the polymerase chain reaction technique. Of the 21 patients 15 had a clinical diagnosis of tropical spastic paraparesis (TSP/HAM), 5 were asymptomatic relatives, and 1 patient had leukemia. In situ hybridization was positive in samples from 5 patients; 2 of these were TSP/HAM patients and the other 3 were healthy wives of TSP/HAM patients. This study demonstrates for the first time that viral RNA is expressed in uncultured PBMNC's of some patients with TSP/HAM in whom proviral DNA is also present; furthermore, the detection of HTLV‐I RNA in the blood of female partners of TSP/HAM patients clearly illustrates the high likelihood of HTLV‐I transmission through sexual contact.
Intrathecal IgG synthesis and specificity of oligoclonal IgG in patients infected with HIV‐1 do not correlate with CNS disease
Authors: Goswami K. K. , Kaye S. , Miller R. , McAllister R. , Tedder R.
Published: March 2007

The CSF/serum immune response to HIV 1 was studied in 24 patients admitted for investigation. The level of antibody to HIV‐1 and specificity of oligoclonal IgG were determined in blood and cerebrospinal fluid (CSF). The majority of patients demonstrated elevated levels of intrathecal IgG synthesis, with levels of HIV‐l‐specific antibody frequently being significantly higher in CSF than in serum. In 16 of 21 patients the CSF/serum antibody ratio indicated active intrathecal synthesis. Oligoclonal banding was present in CSF from all 24 patients. lmmunoprinting of serum and CSF demonstrated antigenic specificity (p24, gp 160, RT) of the clonal antibodies in all of 12 patients though the patterns of reactivity in CSF did not necessarily correspond with that of serum. Although a specific association of particular patterns with HIV CNS disease was not found we feel that these markers should be included in longitudinal studies of HIV‐related diseases of the CNS. The specificity of oligoclonal antibodies, both in CSF and in serum was demonstrated, and this specificity may be a useful marker for longitudinal studies in HIV‐1 antibody‐positive asymptomatic patients.
Measles virus specific antibody in infants in a highly vaccinated society
Authors: Chui Linda W‐L. , Marusyk Raymond G. , Pabst Henry F.
Published: March 2007

Measles virus specific antibody levels were measured in infants from 2 to 12 months of age. The sera were tested by hemagglutination‐inhibition (HI), neutralization (NT), and enzyme immunoas‐say (EIA) methods. The results of this study indicate that in the population examined, infants at an early age have very low or no immunity of maternal origin to measles virus‐93% of the infants were without detectable neutralizing antibody (NT liter ≤10) at 6 months of age, and by the end of the first year of life 100% had no neutralizing antibody.
Stability of molluscum contagiosum virus DNA among 184 patient isolates: Evidence for variability of sequences in the terminal inverted repeats
Authors: Bugert J. J. , Darai G.
Published: March 2007

The stability of the Molluscum contagiosum virus Type 1 genome (188 kbp) was studied in 184 DNA isolates from 131 patients. Variability of up to 1.5 kbp at both ends of the genome symmetrically was observed using restriction analysis of the DNA isolates and by Southern Blot experiments using cloned and labeled Hindlll terminal DNA fragments of MCV‐1 prototype DNA. The variable sequences were mainly confined to the terminal fragments and parts of the MCV‐1 terminal repeats.

Labeled probes did not detect terminal sequences of MCV Type 2 under the applied stringency. A less marked instability of the central MCV‐1 BamHI DNA fragment F was observed within the genome coordinates 0.431 to 0.454 mu. Reiteration of tandem repeats similar to those described for vaccinia virus might explain the variability of the terminal sequences and might be involved in viral replication.
Effect of respiratory syncytial virus infection on mice with protein malnutrition
Authors: Peña‐Cruz Victor , Reiss Carol , McIntosh Kenneth
Published: March 2007

Respiratory syncytial virus (RSV) pulmonary infection was produced in BALB/c mice fed protein‐deficient diets in an effort to understand the severity of viral pneumonia in infants in developing countries. As in previously published experiments with Sendai virus, animals on the deficient diet became clinically malnourished, and certain aspects of their cell‐mediated immunity were altered. The course of RSV infection in protein‐deprived mice was essentially identical to that in normally nourished animals. The titer of virus recovered from lung homogenates over time, as well as the histologic picture of bronchiolitis, were identical under all experimental conditions. This model, unlike that of Sendai virus infection, fails to demonstrate an effect of protein malnutrition on RSV infection.
Recognition of the N‐terminal, C‐terminal, and interior portions of HBx by Sera from patients with hepatitis B
Authors: Kay Alan , de Dinechin Stéphane Dupont , Vitvitski‐Trépo Ludmila , Mandart Elisabeth , Shamoon Blanche‐Marie , Galibert Francis
Published: March 2007

We have cloned and expressed in Escherichia coli three different parts of the HBx open reading frame, the N‐ and C‐termini and the interior or central portion, using two vector systems. The sera of 43 hepatitis B virus patients representing three clinical categories–asymptomatic carriers, chronic active hepatitis, and hepatitis B patients with cirrhosis–known to be anti‐HBx positive, were tested for reactivity against these constructs by Western blotting. The great majority of sera, regardless of the clinical categories, clearly recognise all three parts of HBx, strongly suggesting that the normal mechanism of biosynthesis of the HBx gene product is a straightforward translation of the open reading frame starting from the first ATG. However, asymptomatic carriers show a marked, often almost exclusive, preference for recognition of the central portion of HBx, while patients with chronic hepatitis and patients with cirrhosis generally recognise all three parts of HBx to a similar extent.
Antibody reactivity with two strains of human herpesvirus‐6 in Malaysians
Authors: Yadav M. , Umamaheswari S. , Ablashi D.
Published: March 2007

A total of 234 sera from healthy Malaysians of diverse ethnic origins were tested for antibody to the Z29 and prototype GS strains of HHV‐6. The prevalence in the races ranged from 58 to 80% for the GS strain and 49 to 76% for the Z29 strain. The highest prevalence was in Malays with semi‐urban cultural lifestyles and lowest was in the indigenous rural tribes (Ibans, Kadazans, Bidayuhs, and Orang Asli).

The antibody litres to GS and Z29 virus capsid antigens differed in 11 (4.7%) samples by more than 2 dilutions. In 9 of the 11 sera the titres to GS strain were higher than to the Z29 strain. The differences in the antibody titres between strains of HHV‐6 may reflect subtle changes in antigen structure of the virus recognised by some individuals.
Detection of hantavirus RNA in tissues of experimentally infected mice using reverse transcriptase‐directed polymerase chain reaction
Authors: Xiao Shu‐Yuan , Yanagihara Richard , Godec Mark S. , Eldadah Zayd A. , Johnson Bruce K. , Gajdusek D. Carleton , Asher David M.
Published: March 2007

Detection of hantaviruses, the etiological agents of hemorrhagic fever with renal syndrome (HFRS), by virus isolation using experimental animals or cell culture is time‐consuming. A more rapid but equally specific method is needed. We used a reverse transcriptase‐directed polymerase chain reaction (RT‐PCR) to detect hantavirus genomic sequences and compared its sensitivity with conventional virus isolation. RNA, extracted by the guanidinium isothiocyanate‐cesium chloride method from hantavirus‐infected Vero E6 cells and from tissues of infant mice inoculated intra‐cerebrally with 100 LD50 of hantavirus, was initially reverse transcribed using avian myeloblas‐tosis virus reverse transcriptase. The resulting complementary DNA (cDNA) was used as template to amplify the glycoprotein 2‐encoding region of the hantavirus M segment. With this method, Vero E6 cell cultures infected with Han‐taan virus strains 76–118 (prototype) and HV114 (an isolate from the urine of an HFRS patient in China) were positive, while control cultures were negative. Brain, lung, and heart tissues from hantavirus‐infected mice were positive by RT‐PCR at 5, 8, and 11 days after intracerebral inoculation. The specificity of the positive results was confirmed by restriction endonuclease digestion of the amplified fragments with Alul and Hpal. The sensitivity of the RT‐PCR was equal to cell culture amplification but required less time. This method is being adapted for detection of hantavirus genomic sequences in clinical specimens and postmortem tissues from patients with HFRS.
Localization of rhinovirus replication in vitro with in situ hybridization
Authors: de Arruda Eurico , Mifflin Theodore E. , Gwaltney Jack M. , Winther Birgit , Hayden Frederick G.
Published: March 2007

To facilitate understanding of human rhinovirus (HRV) pathogenesis, methods were developed for detection of HRV infection in vitro using in situ hybridization (ISH). HRV‐14 RNA probes and oligonucleotide probes representing conserved sequences in the 5′‐non‐translated region were labeled with 35S and used to detect infected HeLa or WI‐38 strain human embryonic lung cells in cytological preparations. ISH was shown to be specific for detection of HRV on a single‐cell basis. Subsequently, in human nasal polyps infected in vitro, both oligonucleotide‐ and riboprobes produced a strong signal in association with ciliated epithelial cells. In human adenoids infected in vitro, a signal was observed in nonciliated epithelial cells. This study shows that HRV replicates in ciliated cells in the epithelium of human nasal polyps infected in vitro, and the presence of viral RNA in non‐ciliated cells of the human adenoid infected in vitro suggests that other cell types may also support rhinovirus replication.
Demonstration of concurrent dengue 1 and dengue 3 infection in six patients by the polymerase chain reaction
Authors: Laille Manola , Deubel Vincent , Sainte‐Marie François Flye
Published: March 2007

A dual viremia resulting from naturally acquired dengue 1 and dengue 3 infections in six patients experiencing a dengue‐like syndrome during the epidemic in New Caledonia in 1989 is reported. Serotype identification was first based on virus isolation in mosquito cells and immunofluorescence using type‐specific monoclonal antibodies. The double infection was confirmed directly in blood samples by the polymerase chain reaction (PCR) on genomic RNA and hybridization of the amplified cDNA fragments with type‐specific DNA probes.
Neutralizing antibody response to cytomegalovirus in seropositive pregnant women
Authors: Tanaka Akira , Moriuchi Hiroyuki , Hirota Kiyokata , Numazaki Yoshio
Published: March 2007

The neutralizing (NT) antibody response to human cytomegalovirus (HCMV) in seropositive pregnant women was evaluated to establish the role of protective humoral immunity in HCMV infections (reactivation) during pregnancy. Complement fixation (CF) antibody titers increased significantly in 54 (0.62%) of 8,663 seropositive cases tested during gestation. However, titers of NT antibody, when tested in the presence of complement (C′), increased as gestation progressed and exceeded 1:500 in most of the cases in whom the CF antibody titer had shown earlier an insignificant increase. When NT antibodies were tested in the absence of C′, insignificant increase in titers were found. The results suggest that the presence of high titer of NT antibody may play an important role in protective humoral immunity against reactivation of HCMV in pregnant women, and may also be effective in preventing intrauterine infection and modulating the severity of perinatal infection in offspring borne of HCMV carrier mothers.
Chronic delta hepatitis in haemophiliacs
Authors: Gerritzen Andreas , Brackmann Hans , Van Loo Birgit , Nies Gudrun , Ruland Christiane , Spiegelberg Gertrud , Hammerstein Ursula
Published: March 2007

The seroepidemiological profile of HBV and HDV was investigated in 640 male haemophiliacs. Twenty‐seven of forty‐four HBsAg carriers were anti‐HDV‐lgG positive, 22 were also anti‐HDV‐IgM positive. A markedly lower prevalence of HDV infection was found in patients with anti‐HBc in the absence of HBsAg and anti‐HBs (6/41). Repeated detection of anti‐HDV‐IgM in 5/41 individuals of this group indicates that circulating HBsAg is not an absolute prerequisite for chronic HDV infection.

Overall, chronically active HDV infection was detected more frequently in quiescent than in active chronic HBV infections. Anti‐HDV‐lgM was not detected in the absence of anti‐HDV‐lgG antibodies. Anti‐HDV‐IgG may disappear after resolution of HDV infection, as indicated by the low prevalence (1/42) in such individuals with past HBV infection as well as by loss of anti‐HDV‐lgG observed in two patients.
Hepatitis delta virus cDNA sequence from an acutely HBV‐infected chimpanzee: Sequence conservation in experimental animals
Authors: Kos Ton , Molijn Anco , Van Doorn Leen‐Jan , Van Belkum Alex , Dubbeld Martin , Schellekens Huub
Published: March 2007

Hepatitis delta virus (HDV) RNA was isolated from the serum of a chimpanzee acutely infected with hepatitis B virus (HBV) and superinfected with HDV. Interference of HDV with HBV resulted in decreased HBV DNA levels in the serum. This interference did not change the size of the two HBV specific RNAs present in the liver of the chimpanzee. The complete cDNA sequence of the HDV RNA (5th passage) was determined. Comparison of this cDNA sequence with our previously published sequence (4th passage), located in the variable domain of HDV, was highly conserved. The HDV strain used for these infections originated from a human HDV isolate also used for five to seven HDV passages in chronic HBV carrier chimpanzees (subtypes adw and ayw) or woodchucks chronically infected with woodchuck hepatitis virus (WHV). The complete HDV cDNA sequence showed an extreme conservation (up to 99.8% homology) with the previously published animal‐derived HDV cDNA sequences irrespective of passage number and animal species. In contrast a markedly lower homology (85–89%) was found when compared with 3 human‐derived HDV cDNA sequences. Comparison of our complete cDNA sequence with the human‐derived cDNA sequences showed that the nucleotide changes in the human‐derived isolates were restricted to specific regions on the genome and to specific basepair substitutions. The hepatitis Delta antigen (HDAg) is highly conserved both in the human‐ and animal‐derived cDNA sequences showing mainly conservative amino acid changes.
Human parvovirus B19 specific IgG, IgA, and IgM antibodies and DNA in serum specimens from persons with erythema infectiosum
Authors: Erdman Dean D. , Usher M. Jane , Tsou Cecilia , Caul E. Owen , Gary G. William , Kajigaya Sachiko , Young Neal S. , Anderson Larry J.
Published: March 2007

To determine the diagnostic use of different markers of acute parvovirus 619 infection, serum specimens obtained from 128 persons with erythema infectiosum were tested for specific immunoglobulin G (IgG), IgA, and IgM antibodies by capture enzyme immunoassay (EIA) using Chinese hamster ovary (CHO) cell‐expressed B19 antigen, and tested for circulating B19 DNA by polymerase chain reaction (PCR). A significant rise in specific IgG and IgA antibodies was detected in 87% and 77%, respectively, of persons from whom acute‐ and convalescent‐phase serum specimens were available. Specific IgA antibodies were detected in single serum specimens from 90% of cases and were present in 22 (18%) of 120 persons from a control group without a history of recent exposure to B19. Specific IgM antibodies were detected i n 97% of cases and one person (IYO) from the control group. B19 DNA was detected in 94% of cases and was absent in 20 persons from the control group positive for both IgG and IgA antibodies. Serum specimens obtained between 4 and 6 months after onset of illness from six additional persons were also tested. All had specific IgG antibodies, four (67%) had IgA, five (83%) had IgM, and none had detectable B19 DNA. Our data indicate that 1) specific IgA antibodies are too persistent to be a useful indicator of recent B19 infection; 2) specific IgM antibodies are the most sensitive indicator of acute B19 infection in immunologically normal persons but can persist up to 6 months; and 3) 619 DNA can often be detected up to 2 months after onset of illness even in immunologically normal hosts and might be a useful adjunct test for diagnosis of acute B19 infection.
Diagnosis of acute and latent varicella‐zoster virus infections using the polymerase chain reaction
Authors: Dlugosch Dorothea , Eis‐Hübinger Anna M. , Kleim Jörg‐P. , Kaiser Rolf , Bierhoff Erhard , Schneweis Karl E.
Published: March 2007

A simplified assay for the diagnosis of varicellazoster virus (VZV) infections based on the polymerase chain reaction (PCR) is described. Omitting the procedures for extraction and purification of DNA, the crude vesicle fluid materials were used for PCR. Moreover, hybridization was not necessary for detection of the amplification products because they were already visible after ethidium bromide staining of the electrophoresis gel. Results could therefore be obtained within one day. In comparison to virus isolation, PCR proved much more rapid, highly sensitive, and specific. DNA extraction and a double PCR assay with nested primers were necessary for detection of latent VZV infections in trigeminal and thoracic ganglia. The data suggest that the procedures described are universally applicable to several types of specimens dependent on the calculated amount of target DNA.
A prospective study of in vitro anti‐HBs producing B cells (spot‐ELISA) following primary and supplementary vaccination with a recombinant hepatitis B vaccine in insulin dependent diabetic patients and matched controls
Authors: Wismans P. J. , Van Hattum J. , De Gast G. C. , Bouter K. P. , Diepersloot R. J. A. , Maikoe T. J. , Mudde G. C.
Published: March 2007

A prospective study of the immune response after hepatitis B vaccination was carried out in 32 insulin dependent diabetes mellitus (IDDM) patients and their age and sex matched healthy controls. A sensitive, immunoenzymatic technique was used, able to detect in vitro specific antibody production by mitogen stimulated individual B cells.

In‐vivo serologic response after vaccinationwith a standard scheme (0, 1 and 6 months) of 20 pg recombinant hepatitis B (HB) vaccine was significantly impaired in the IDDM patients both with respect to the number of nonresponders (25 versus 3%, P < 0.05) and antibody titers reached (1,377 vs. 9,060 IU/L, P < 0.05). The total number of in vitro IgM‐ and IgG‐class immunoglobulin producing B cells as detected by the spot‐ELISA, was found to be comparable in both groups. Specific IgG anti‐HBs (and to a lesser extent IgM anti‐HBs) showed impairment in the diabetic population as a whole. The number of IgG anti‐HBs producing B cells was mark‐ edly depressed one month following vaccination, which is probably a reflection of homing of B cells outside the circulation.

Responding subjects were identified early during their vaccination by the detection of in vitro anti‐HBs production using the spot‐ELISA. Nonresponding healthy subjects and IDDM patients as a group showed a low number of IgG anti‐HBs spots, suggesting a reduced specific memory B cell frequency. In 13 of 15 hypo‐ and nonresponders with positive IgG anti‐HBs spots supplementary vaccination(s) resulted in improved anti‐HBs levels.
Direct isolation of inoue‐melnick virus from human colorectal carcinoma tissues but not from stomach carcinoma tissues
Authors: Inoue Y. K. , Nishibe Y. , Nakamura Y.
Published: March 2007

Inoue‐Melnick virus (IMV) was isolated directly from all of the seven human colorectal adenocarcinoma tissues studied, whereas IMV was not isolated from five human stomach carcinoma tissues. Seven isolates from the Japanese colorectal carcinoma tissues were identified as type 1 IMV by the neutralization test and were sensitive to phosphonoacetic acid, a growth inhibitor of IMV. Furthermore, type 1 IMV‐antibody was detected in six of seven sera of the patients whose colorectal carcinoma tissues were positive for virus isolation. In contrast, IMV‐antibody was not detected in the five sera of the patients whose stomach carcinoma tissues were tested and found negative for IMV.
Frequent isolation of intertypic poliovirus recombinants with serotype 2 specificity from vaccine‐associated polio cases
Authors: Lipskaya Galina Y. , Muzychenko A. R. , Kutitova Olga K. , Maslova Svetlana V. , Equestre M. , Drozdov S. G. , Bercoff R. Perez , Agol V. I.
Published: March 2007

Five representatives from a collection of 21 Sabin type 2‐like poliovirus strains isolated from paralytic poliomyelitis cases in two regions of the USSR have been subjected t o limited nucleotide sequencing. All proved to be intertypic recombinants having the genes encoding capsid proteins of Sabin 2 origin and a 3'‐end portion of the genome derived from either type 1 (3 isolates) or type 3 (2 isolates) Sabin strains. The crossover points in all the 5 genomes have been mapped to different loci of the P3 region. At least 6 additional isolates from the same collection (and 2 isolates from healthy contacts), appeared to have a type 2/type 1 recombinant genome, as judged by oligonucleotide mapping. The biological significance of frequent occurrence of recombinants among field isolates of vaccine‐related strains is discussed.