bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of enteric adenoviruses by dot‐blot hybridization using a molecularly cloned viral DNA probe
Authors: Takiff Howard , Seidlin Mindell , Krause Philip , Rooney James , Straus Stephen E. , Brandt Carl , Rodriguez William , Yolken Robert 
Published: March 2007

AbstractEnteric adenoviruses (EAds) (candidate adenoviruses 40 and 41, subgroups F and G) have been implicated in the etiology of gastroenteritis in infants, but their clinical significance has been unclear because a rapid test to distinguish these agents from other adenovirus (Ad) types has not been available. We developed a dot‐blot hybridization assay for EAd DNA using a cloned DNA fragment that has little homology to non‐EAd DNAs. The dot‐blot system detected less than 20 pg of EAd DNA, while showing minima cross hybridization to representative strains from all other Ad groups. There was no detectable hybridization to extracts of samples known to contain other enteric viruses. It was further shown that low levels of EAds in specimens could be amplified by culturing for 1 day in 293 cells. Stool samples and tissue culture lysates prescreened by electron microscopy, cell culture or ELISA were tested in a blind fashion. Using endonuclease analysis as the standard for typing the isolates, we found the dot‐blot system to have a 91% sensitivity and 71% specificity for detecting EAds and distinguishing them from other Ads. False‐positive and equivocal dot‐blot results appeared to be caused by other Ads.
Detection of adenoviruses in stool specimens by nucleic acid spot hybridization
Authors: Stålhandske Per , Hyypiä Timo , Allard Annika , Halonen Pekka , Pettersson Ulf 
Published: March 2007

AbstractNucleic acid hybridization was used for the detection of adenovirus DNA in stool specimens, and the results were compared with those obtained by a radioimmunoassay (RIA) for adenovirus hexon antigen. DNA from 40 specimens, 18 of which were positive by RIA, were spotted onto nitrocellulose filters and analyzed by hybridization using radioactively labeled adenovirus‐2 DNA or a cloned DNA fragment from enteric adenovirus‐41 as probes. With the adenovirus‐2 DNA probe, 15 of the 18 RIA‐positive specimens were also positive in the hybridization assay, and one of the RIA negative specimens was also scored as positive. The cloned adenovirus‐41 fragment gave a positive signal with five specimens, all of which were also detected with the adenovirus‐2 DNA probe. The results show that hybridization is an alternative method for detection of adenovirus in stool specimens. The sensitivity of the assay is comparable to that of the RIA.
Natural killing (NK) of cytomegalovirus (CMV)‐infected fibroblasts: A comparison between two strains of CMV, uninfected fibroblasts, and K562 cells
Authors: Waner Joseph L. , Nierenberg Julia A. 
Published: March 2007

AbstractSpontaneous cytolysis of uninfected human fibroblasts (HF), K562 cells, and HF cells infected with cytomegalovirus (CMV) was associated with nonadherent peripheral blood leukocytes (PBL) that passed through a nylon wool column, and rosetted with sheep erythrocytes at low affinity; cytolysis was further associated with enriched preparations of large granular lymphocytes (LGL). The Leu‐7 marker did not correlate as a cell phenotype with functional activity in normal donors. The HF cells infected with the AD169 strain of CMV were as sensitive to cytolysis as K562 cells, although the kinetics of cytolysis differed; HF cells infected with the Davis strain of CMV were generally less sensitive to cytolysis than uninfected fibroblasts.Equivalent amounts of interferon were detected in NK assays containing Davis targets, AD169 targets, or K562 cells; interferon was not detected in NK assays of HF cells. Neither uninfected nor infected HF cells competed effectively against51Cr‐labeled K562 cells in cold competition experiments; K562 cells, however, competed effectively against51Cr‐labeled AD169‐CMV targets. Uninfected HF cells were efficient competitors of AD169‐CMV targets. Davis‐CMV‐infected cells were less effective, however, than uninfected HF cells in competition experiments, suggesting that fewer receptors for NK cells were present on these targets.
Immunostimulatory function of herpes simplex virus isolates from patients with frequent herpes labialis and a deficiency in immunespecific interferon production
Authors: Klieman Rachel L. , Green Jon A. , Spruance Spotswood L. 
Published: March 2007

AbstractApproximately 30% of persons with frequent episodes of herpes labialis are deficient in the production of HSV‐induced immunespecific interferon (IFN) (Green, 1985). Herpes simplex virus (HSV) strains isolated from persons who make immunespecific IFN and from persons who do not make it were examined for their immunostimulatory capabilities. HSV isolated from the primary oral lesions of two patients deficient in immunespecific IFN production, one person with an intact immunespecific IFN response, HSV types 1 and 2 laboratory strains, and Newcastle disease virus (NDV) were added to cultures of peripheral blood mononuclear cells (PBML) from HSV seropositive donors. All HSV‐isolates induced comparable titers of immunespecific IFN. These studies suggest that failure of some patients to develop an immunespecific IFN response is determined by the host, not the virus.
Detection of RS‐virus IgG‐subclass‐ specific antibodies: Variation according to age in infants and small children and diagnostic value in RS‐virus‐infected small infants
Authors: Hornsleth Allan , Bech‐Thomsen Niels , Friis Birgitte 
Published: March 2007

AbstractThe variation according to age of RS‐virus IgG‐subclass specific antibodies was investigated by ELISA in 140 acute phase sera from 140 infants and children 1–47 mo of age who were hospitalized with acute respiratory disease. Sixty‐nine of these serum samples were obtained from patients with RS‐virus infections. Mean OD values of IgG‐1 antibodies decreased corresponding to the decrease in maternal antibodies. In patients with RS‐virus infections, the mean IgG‐1 OD values increased in children who were 3 yr old, but in patients without RS‐virus infections this increase was observed in children at 1 yr of age. RS‐virus IgG‐3 antibodies were detected only in children 1–3 yrs of age, and only in 18% of samples from children without RS‐virus infections. The concept of the transient nature of RS‐virus IgG‐3 antibodies as compared with RS‐virus IgG‐1 antibodies is supported by these results.RS‐virus subclass specific antibodies were studied in paired serum samples from 12 infants 1–5 mo of age with acute lower RS‐virus disease. IgG‐1 antibodies developed in six infants, but only in one of six infants aged 1–3 mo. IgG‐3 antibodies developed in nine infants, including four of six infants who were 1–3 mo old. These results suggest that in small infants with RS‐virus infections the detection of IgG‐3 antibodies is of higher diagnostic value than the detection of IgG‐1 antibodies.
Detection of BK virus antibodies measured by enzyme‐linked immunosorbent assay (ELISA) and two haemagglutination inhibition methods: A comparative study
Authors: Flaegstad Trond , Traavik Terje 
Published: March 2007

AbstractWe developed a solid phase enzyme‐linked immunosorbent assay (ELISA) for detecting IgG antibodies to BK virus (BKV). This test was compared to two haemagglutination inhibition methods, one using receptor‐destroying enzyme and the other NaIO4 to remove nonspecific inhibitors. The results show that this single dilution ELISA is a sensitive and specific test. The inherent advantages of this method make it an attractive alternative for the seroepidemiology of BKV.
Detection of cytomegalovirus in urine samples by enzyme‐linked immunosorbent assay
Authors: McKeating J. A. , Stagno S. , Stirk P. R. , Griffiths P. D. 
Published: March 2007

AbstractAn enzyme‐linked immunosorbent assay (ELISA) was developed for the detection of cytomegalovirus (CMV) in urine using monoclonal antibodies directed against CMV as a capture for viral antigen. The assay was capable of detecting virus at 102.3TCID50/ml as determined by titration of stock virus, strain Ad169.The assay was found to have a sensitivity of 65% and a specificity of 100% when 73 coded stored urine specimens were examined. Assuming that the poor sensitivity was due to loss of antigen following storage, we proceeded to analyse fresh urine specimens. Surprisingly, the assay gave negative results with 46 fresh urines known to contain CMV; however, following storage at +4°C for two weeks, 35 (76%) of these samples gave ELISA results in the positive range. This detection of CMV, after storage at +4°C, could be due to degradation of virus particles leading to release of soluble glycoproteins into the medium or to the presence of an inhibitory substance in fresh urine that is destroyed during storage.
Relative frequency of human rotavirus subgroups 1 and 2 in Japanese children with acute gastroenteritis
Authors: Nakagomi Osamu , Nakagomi Toyoko , Oyamada Hajime , Suto Tsunehisa 
Published: March 2007

AbstractRecently developed monoclonal antibodies against the 42,000‐dalton major inner capsid protein were used in an enzyme immunoassay to subgroup a total of 156 rotavirus specimens obtained from Japanese infants and young children with acute gastroenteritis during the period between December 1981 and April 1983. Only 2 specimens (1.3%) were identified as subgroup 1, whereas 154 specimens (98.7%) were identified as subgroup 2. The percentage of subgroup 2 rotaviruses obtained in this study was the highest among similar studies thus far performed in different areas of the world. One of the subgroup 1 isolates had fast‐moving 10th and 11th gene segments (the “long” pattern) typical of the subgroup 2 human rotaviruses.
Human antibody response to immunization with 17D yellow fever and inactivated TBE vaccine
Authors: Kayser M. , Klein H. , Paasch I. , Pilaski J. , Blenk H. , Heeg K. 
Published: March 2007

AbstractThe antibody response against flaviviruses tick‐borne encephalitis (TBE), Kyasanur Forest disease (KFD), Murray Valley encephalitis (MVE), West Nile fever (WNF), Japanese B encephalitis (JE), dengue 2 (DEN‐2), and yellow fever (YF) was studied in humans after administration of an inactivated TBE virus vaccine. Individuals were either prevaccinated with 17D yellow fever (experimental group) or without any previous exposure to flaviviruses (control group). The appearance of serum titres of homologous and heterologous haemagglutination inhibition (HI) antibodies, heterotypic DEN‐2 neutralizing antibodies, and TBE enzyme‐linked immunosorbent assay (ELISA) antibodies were examined.Individuals prevaccinated with the 17D yellow fever developed an antibody pattern that contrasted with that of the control group. This pattern was characterized as follows: (1) Predominantly anti‐TBE IgG antibodies appeared earlier and in higher titres than in the control group, (2) heterologous HI antibodies crossreacting with the WN flavivirus subgroup preceded the appearance of homologous HI antibodies, (3) a broad spectrum HI response was observed against all flaviviruses tested, and (4) low titre heterotypic DEN‐2 neutralizing antibodies were formed in about half of the cases. These observations are discussed in the context of cross‐reactivity, cross‐protection and virus infection enhancement.
Genetic relatedness among human rotaviruses
Authors: Flores Jorge , Perez‐Schael Irene , Boeggeman Elizabeth , White Laura , Perez Mireya , Purcell Robert , Hoshino Yasutaka , Midthun Karen , Chanock Robert M. , Kapikian Albert Z. 
Published: March 2007

AbstractThe genetic relatedness of 81 clinical rotavirus isolates to the human rotavirus prototype strains Wa (subgroup 2, serotype 1) and DS‐1 (subgroup 1, serotype 2) was examined by RNA hybridization techniques. Labeled single‐stranded ( + ) transcripts of Wa or DS‐1 virus were incubated with denatured genomic rotaviral RNAs, and the resulting hybrids were subjected to gel electrophoresis and autoradiography. Nineteen of the specimens contained subgroup 1 rotavirus with a “short” RNA migration pattern. These viruses were found to be closely related to the DS‐1 strain and were associated with illness of short duration. The remaining 62 isolates belonged to subgroup 2 and exhibited a “long” RNA migration pattern. Fifty‐four of these isolates exhibited significant hybridization with the Wa strain probe. Four isolates yielded multiple hybrid bands with the Wa probe but also possessed at least one gene segment homologous to the DS‐1 strain. The remaining four subgroup 2 rotaviruses did not exhibit significant homology in the form of labeled hybrid bands when tested with either the Wa or DS‐1 probe. These findings suggest that most clinical rotavirus isolates belong to one of two human rotavirus “families” defined as Wa‐like or DS‐1‐like. Our observations also suggest that reassortment occurs in vivo between rotaviruses belonging to the two human rotavirus “families” and that there are one or more additional families of human rotavirus.
Relationship between HBcAg in serum and liver and HBV replication in patients with HBsAg‐positive chronic liver disease
Authors: Pontisso Patrizia , Chemello Liliana , Fattovich Giovanna , Alberti Alfredo , Realdi Giuseppe , Brechot Christian 
Published: March 2007

AbstractThe expression of hepatitis B core antigen (HBcAg) in serum and in hepatocytes was evaluated in relation to HBV replication. Fifty chronic HBsAg carriers with histological evidence of liver disease were studied, including 24 HBeAg‐positive patients, 2 HBeAg/anti‐HBe‐negative patients, and 24 anti‐HBe‐positive cases, two of them with evidence of delta agent infection. Serum HBV‐DNA was evaluated in all patients and related to HBcAg examined at the same time in frozen liver biopsies by immunofluorescence and to HBcAg detected in the corresponding serum by a recently developed radioimmunoassay.HBV‐DNA was present in serum in 20 (83%) HBeAg‐positive patients, all positive for serum HBcAg, whereas liver core antigen was detected in 14 (73%) of 19 cases. Among HBeAg‐negative patients, 50% showed the presence of circulating DNA viral sequences, and HBcAg was identified in five of 26 (19%) cases in serum and in six of 24 (25%) in the liver respectively. In 15 patients, liver fragments permitted examination in parallel by immunofluorescence for HBcAg and molecular hybridization for viral DNA in liver cells. A DNA pattern characteristic of viral replication was found in cases with evidence of active virion production, independently from HBeAg and anti‐HBe, and in these patients HBcAg was present both in serum and in hepatocytes. In two cases with free HBV‐DNA, without evidence of replicative activity, core antigen was not detected in the liver, but in one patient HBcAg was found in the serum. A similar finding was also noted in another patient, in whom the hybridization pattern was consistent with integration of viral genome into high‐molecular‐weight cellular DNA. Whether serum HBcAg detected in these patients without HBV‐DNA in serum reflects the presence of defective viral particles or of core antigen released as a viral protein remains to be determined.
Detection of BK virus IgM antibodies by two enzyme‐linked immunosorbent assays (ELISA) and a hemagglutination inhibition method
Authors: Flaegstad Trond , Traavik Terje 
Published: March 2007

AbstractWe have used an antigen solid‐phase enzyme‐linked immunosorbent assay (SP‐ELISA) and an IgM antibody capture ELISA (MACELISA) for detecting IgM antibodies to human polyomavirus BK (BKV). These tests were compared with the standard hemagglutination inhibition test (HAI) of IgM serum fractions following sucrose density gradient fractionation. The SP‐ and MACELISA were not influenced by concomitant BKV‐IgG, but high levels of both BKV‐IgG and rheumatoid factor could cause false positive results by SPELISA, but not by MACELISA. The MACELISA gave much higher positive to negative ratios than the SPELISA.The sensitivity and specificity of the two tests were high compared to the IgM‐HAI method. The sera could be tested in a single dilution (1:160), and thus the ELISA‐tests are useful for testing large numbers of sera.
Identification of genital tract papillomaviruses HPV‐6 and HPV‐16 in warts of the oral cavity
Authors: Naghashfar Zohreh , Sawada Edward , Kutcher Mark J. , Swancar James , Gupta Jean , Daniel Richard , Kashima Haskins , Woodruff J. Donald , Shah Keerti 
Published: March 2007

AbstractWarty lesions of the oral cavity were examined for etiologic association with genital tract papillomaviruses HPV‐6, HPV‐11, and HPV‐16. DNAs extracted from ten oral biopsies were screened for HPV genomic sequences by Southern transfer hybridization with 32P‐labeled viral DNA probes. Nonstringent hybridization with an HPV‐6 probe revealed papillomavirus DNA sequences in four of seven tissues with histologic evidence of papillomatosis, in none of two tissues without histologic evidence of papillomatosis, and in one tissue that was not examined by histology. Stringent hybridization tests with HPV‐6 and HPV‐16 probes identified the genome in one tissue as being HPV‐16, in a second tissue as being HPV‐6 subtype a, and in a third tissue as HPV‐6 (subtype unidentified); papillomavirus DNA sequences in two tissues are as yet not identified. An additional case of HPV‐6 or HPV‐11 related oral cavity lesion was diagnosed by in situ hybridization of paraffin sections with a 35S‐labeled, mixed HPV‐6 + HPV‐11 probe. The hybridization in the positive section was extensive and confined to epithelial nuclei. The oral lesions associated with genital tract papillomaviruses were asymptomatic, multiple or single, and were located in different parts of the oral cavity, for example, on the gingivae, on the tongue, on the lip, on the tonsillar pillar, and on the floor of the mouth.
Comparison of long‐term systemic and secretory antibody responses in children given live, attenuated, or inactivated influenza A vaccine
Authors: Johnson Philip R. , Feldman Sandor , Thompson Juliette M. , Mahoney Josephine D. , Wright Peter F. 
Published: March 2007

AbstractA comparison of inactivated intramuscular and live intranasal influenza A vaccines in young children undergoing primary immunization might be expected to show differences in serum and local mucosal antibody responses. To demonstrate such differences, serum and local respiratory tract antibody responses of young children vaccinated with intranasal live, attenuated, cold‐adapted (H3N2 or HINI), or intramuscular inactivated (H3N2) influenza A vaccines were examined for one year after vaccination. Antibody responses were measured by hemagglutination inhibition (HAI) and class‐specific enzyme‐linked immunosorbent assay (ELISA). One year after vaccination, live intranasal vaccinees had significantly less decay of serum HAI (p = 0.025) and IgG antibody (p = 0.01) directed against the influenza hemagglutinin and neuraminidase than did intramuscular inactivated vaccinees. Nasal secretory IgA developed almost exclusively in live vaccinees and persisted for up to one year. Persistent nasal secretory IgG was detected in both live and inactivated vacinees. Live vaccination not only stimulates a more durable serum antibody response, but also induces long‐lasting local respiratory tract IgA antibody that may play an important role in host protection.
Cellular immune responses in mice challenged with an amyocarditic variant of coxsackievirus B3
Authors: Lutton C. W. , Gudvangen R. J. , Nealon T. J. , Paque R. E. , Gauntt C. J. 
Published: March 2007

AbstractAn amyocarditic variant of a temperature‐sensitive (ts) mutant derived from the parent myocarditic variant Coxsackievirus B3 (CVB3m) was studied in a murine model of CVB3m‐induced myocarditis to assess virus‐induced antigens and their possible role in the disease process. Amyocarditic variant ts5R induced a heart tissue antigen(s), extractable by hypertonic KCI, which inhibited migration of peritoneal exudate cells from CVB3‐inoculated myocarditic mice in an agarose droplet cell‐migration‐inhibition assay. The ts5R variant was amyocarditic at inoculum doses of 103 to 108 plaque‐forming units per mouse, but in cyclophosphamide‐immunosuppressed mice, ts5R induced myocarditis. Viable ts5R served as a vaccine and protected mice against CVB3m‐induced myocarditis. Murine neonatal skin fibroblasts (MNSF) infected with either virus served as in vitro targets and were lysed by splenic cytotoxic T lymphocytes from mice inoculated with either virus variant. ts5R and CVB3m replicated to similar titers in murine neonatal skin fibroblasts (MNSF) at 24 hr postinoculation (pi), but differences in titers were found by 72 hr pi. Levels of natural killer cell activities in spleens of ts5R‐inoculated mice were slightly lower than in spleens of CVB3m‐inoculated mice at 7 days pi. The data suggest that viral induction of new antigens on target cells and viral induction of specific cytotoxic T lymphocytes that recognize these antigenic changes do not always result in induction of myocarditis.
Oral bromovinyldeoxyuridine therapy for herpes simplex and varicella‐zoster virus infections in severely immunosuppressed patients: A preliminary clinical trial
Authors: Tricot G. , De Clercq E. , Boogaerts M. A. , Verwilghen R. L. 
Published: March 2007

AbstractTwenty‐five patients with haematological diseases were treated orally with the highly potent and selective anti‐herpes agent, bromovinyldeoxyuridine (BVDU), in a dosage of 7.5 mg/kg/day (divided over three or four doses a day) for 5 days for an intercurrent mucocutaneous herpesvirus infection. Of these 25 patients, 8 were severely granulocytopenic at the time of the viral infection, and 12 recently had undergone bone‐marrow transplantation; 5 were under cytotoxic therapy for a lymphoproliferative disorder; 13 had herpes simplex virus type 1 (HSV‐1); 1 had herpes simplex virus type 2 (HSV‐2); and 11 had varicella‐zoster virus (VZV) infection. In all but two patients, BVDU arrested progression of the HSV or VZV infection within 1–2 days after treatment was started. One of the two patients who failed to respond to BVDU had an HSV‐2 infection. The other had an HSV‐1 infection, which was highly sensitive to BVDU in vitro; BVDU may have failed in this patient because of incomplete drug intake or profuse diarrhoea, or both. The results of this preliminary uncontrolled clinical trial suggest that BVDU may be an effective and safe drug for the oral treatment of HSV‐1 and VZV infections in severely immunosuppressed patients.
Analysis of the genome of molluscum contagiosum virus by restriction endonuclease analysis and molecular cloning
Authors: Darai G. , Reisner H. , Scholz J. , Schnitzler P. , de Ruiz H. Lorbacher 
Published: March 2007

AbstractVirions of molluscum contagiosum virus (MCV), a member of the poxviridae, were isolated directly from lesions of individual patients and characterized by restriction enzyme analysis. The comparative analysis of the cleavage patterns and Southern blot hybridization of 14 independently isolated virus samples revealed that MCV isolates can be classified into two different types. The majority of MCV isolated from clinically typical skin lesions (13 of 14) showed similar DNA cleavage patterns and were termed MCV type 1, whereas one isolate derived from a vaginal lesion showed a completely different DNA cleavage pattern and therefore was termed MCV type 2. For detailed investigation of the viral genome, a defined gene library of MCV DNA sequences was established. The Bam HI DNA fragments of the viral genome of MCV type 1 prototype isolate 1/80 was inserted into the bacterial plasmid vector pAT153. With the exception of terminal fragments (fragments A and B) of the viral genome, all other DNA fragments were cloned. All cloned Bam HI DNA fragments were individually identified by digestion of the recombinant plasmid DNA with different restriction enzymes and screened by hybridization of plasmid DNA to viral DNA.