bluuebunny/crossref_metadata_2025 huggingface dataset:
Transduction-Mediated Transfer of Unmarked Deletion and Point Mutations through Use of Counterselectable Suicide Vectors
Authors: Kang Ho Young , Dozois Charles M. , Tinge Steven A. , Lee Tae Ho , Curtiss Roy
Published: July 2002

A challenge in strain construction is that unmarked deletion and nucleotide substitution alleles generally do not confer selectable phenotypes. We describe here a rapid and efficient strategy for transferring such alleles via generalized transduction. The desired allele is first constructed and introduced into the chromosome by conventional allelic-exchange methods. The suicide vector containing the same allele is then integrated into the mutant chromosome, generating a tandem duplication homozygous for that allele. The resulting strain is used as a donor for transductional crosses, and selection is made for a marker carried by the integrated suicide vector. Segregation of the tandem duplication results in haploid individuals, each of which carries the desired allele. To demonstrate this mutagenesis strategy, we used bacteriophage P22H
int
for generalized transduction-mediated introduction of unmarked mutations to
Salmonella enterica
serovar Typhimurium. This method is applicable to any species for which generalized transduction is established.
Mutational Analysis of the TonB1 Energy Coupler ofPseudomonas aeruginosa
Authors: Zhao Qixun , Poole Keith
Published: July 2002

Siderophore-mediated iron transport inPseudomonas aeruginosais dependent upon the cytoplasmic membrane-associated TonB1 energy coupling protein for activity. To assess the functional significance of the various regions of this molecule and to identify functionally important residues, thetonB1gene was subjected to site-directed mutagenesis, and the influence on iron acquisition was determined. The novel N-terminal extension of TonB1, which is absent in all other examples of TonB, was required for TonB1 activity in bothP. aeruginosaandEscherichia coli. Appending it to the N terminus of the nonfunctional (inP. aeruginosa)Escherichia coliTonB protein (TonBEc) rendered TonBEcweakly active inP. aeruginosaand did not compromise the activity of this protein inE. coli. Elimination of the membrane-spanning, presumed membrane anchor sequence of TonB1 abrogated TonB1 activity inP. aeruginosaandE. coli. Interestingly, however, a conserved His residue within the membrane anchor sequence, shown to be required for TonBEcfunction inE. coli, was shown here to be essential for TonB1 activity inE. colibut not inP. aeruginosa. Several mutations within the C-terminal end of TonB1, within a region exhibiting the greatest similarity to other TonB proteins, compromised a TonB1 contribution to iron acquisition in bothP. aeruginosaandE. coli, including substitutions at Tyr264, Glu274, Lys278, and Asp304. Mutations at Pro265, Gln293, and Val294 also impacted negatively on TonB1 function inE. colibut not inP. aeruginosa. The Asp304 mutation was suppressed by a second mutation at Glu274 of TonB1 but only inP. aeruginosa. Several TonB1-TonBEcchimeras were constructed, and assessment of their activities revealed that substitutions at the N or C terminus of TonB1 compromised its activity inP. aeruginosa, although chimeras possessing anE. coliC terminus were active inE. coli.
Multiyear Prospective Study of Intestinal Parasitism in a Cohort of Peace Corps Volunteers in Guatemala
Authors: Herwaldt Barbara L. , de Arroyave Kathleen R. , Wahlquist Susanne P. , de Merida Anna Maria , Lopez Adriana S. , Juranek Dennis D.
Published: July 2002

We conducted a prospective, longitudinal study in a cohort of 36 Peace Corps volunteers (PCVs) in Guatemala to study the incidence and natural history of intestinal parasitic infections during the PCVs' >2-year overseas stay. PCVs collected stool specimens at least monthly and when ill with gastrointestinal symptoms. Of the 1,168 specimens tested, 453 (38.8%) were positive for at least one parasite and 48 (4.1%) were positive for a pathogenic parasite. A median interval of 187 days (range, 14 to 752 days) elapsed before the first documented parasitic infection, and the median intervals from arrival until subsequent infections (e.g., second or third) were >300 days. The PCVs had 116 episodes of infection with 11 parasites, including up to 4 episodes per PCV with specific nonpathogens and
Blastocystis hominis
. The incidence, in episodes per 100 person-years, was highest for
B. hominis
(65), followed by
Entamoeba coli
(31),
Cryptosporidium parvum
(17), and
Entamoeba hartmanni
(17). The PCVs'
B. hominis
episodes lasted 6,809 person-days (28.7% of the 23,689 person-days in the study), the
E. coli
episodes lasted 2,055 person-days (8.7%), and each of the other types of episodes lasted <2% of the person-days in the study. Gastrointestinal symptoms were somewhat more common and more persistent, but not significantly so, in association with pathogen episodes than with
B. hominis
and nonpathogen episodes. Although infections with pathogenic parasites could account for only a minority of the PCVs' diarrheal episodes, the continued acquisition of parasitic infections throughout the PCVs' >2-year stay in Guatemala suggests that PCVs repeatedly had fecal exposures and thus were at risk for infections with both parasitic and nonparasitic pathogens throughout their overseas service.
Molecular Epidemiology of
Scedosporium apiospermum
Infection Determined by PCR Amplification of Ribosomal Intergenic Spacer Sequences in Patients with Chronic Lung Disease
Authors: Williamson Emma C. M. , Speers David , Arthur Ian H. , Harnett Gerald , Ryan Gerard , Inglis Tim J. J.
Published: July 2002

Respiratory tract colonization with
Scedosporium apiospermum
in patients with chronic suppurative lung disease is a significant concern for lung transplantation candidates, since
Scedosporium
infections occurring posttransplantation are usually untreatable. Up to 10% of patients with cystic fibrosis attending our respiratory medicine unit have had
Scedosporium
organisms isolated from sputum samples. We therefore developed a molecular typing method to examine these isolates. Typing by PCR amplification of ribosomal intergenic spacer sequences demonstrated 20 different types from 52 isolates collected from the respiratory medicine unit and elsewhere in Australia. A single common type was isolated from 11 respiratory medicine unit inpatients. Two other types were isolated from more than one source: one from two respiratory medicine unit inpatients and one from two epidemiologically linked nonhuman sources. Multiple isolates were obtained from nine patients. This method demonstrated persistent carriage of isolates of the same type in one patient for 7 months. Two patients showed carriage of isolates with multiple typing patterns within a 3-month period. The high rate of isolation and the predominance of isolates with a single typing pattern from respiratory medicine unit patients may suggest transmission to patients from a source in the unit. There was no epidemiological evidence of direct patient-to-patient spread, and
Scedosporium
organisms were not isolated from dust, soil, or air samples from the unit. The source and route of transmission have yet to be determined.
Epidemiologic Usefulness of Spoligotyping for Secondary Typing of
Mycobacterium tuberculosis
Isolates with Low Copy Numbers of IS
6110
Authors: Cronin Wendy A. , Golub Jonathan E. , Magder Laurence S. , Baruch Nancy G. , Lathan Monica J. , Mukasa Leonard N. , Hooper Nancy , Razeq Jafar H. , Mulcahy Donna , Benjamin William H. , Bishai William R.
Published: July 2002

Restriction fragment length polymorphism (RFLP) analysis of IS
6110
is commonly used to DNA fingerprint
Mycobacterium tuberculosis
. However, low-copy (≤5) IS
6110 M. tuberculosis
strains are poorly differentiated, requiring secondary typing. When spoligotyping was used as the secondary method, only 13% of Maryland culture-positive tuberculosis (TB) patients with low-copy IS
6110
-spoligotyped clustered strains had epidemiologic linkages to another patient, compared to 48% of those with high-copy strains clustered by IS
6110
alone (
P
< 0.01). Spoligotyping did not improve a population-based molecular epidemiologic study of recent TB transmission.
Flagella as a Potential Marker forCampylobacter jejuniStrains Associated with Guillain-Barré Syndrome
Authors: Tsang Raymond S. W. , Figueroa Guillermo , Bryden Louis , Ng Lai-King
Published: July 2002

Campylobacter jejunirecovered from patients with Guillain-Barré syndrome (GBS) in different geographical locations and bearing different heat-labile and heat-stable antigens were found to have identical amino acid sequences in their flagellarflaAshort variable region, suggesting that it may be a potentially useful marker for GBS association.
Comparison of Two Methods for Serotyping
Campylobacter
spp
Authors: McKay Dorothy , Fletcher June , Cooper Peggy , Thomson-Carter Fiona M.
Published: July 2002

Two serotyping schemes (Penner and Laboratory of Enteric Pathogens [LEP]) based on soluble heat-stable antigens were used to analyze 3,788
Campylobacter
sp. isolates. A significant percentage (36.6%) was untypeable using LEP serotyping; greater cross-reaction was also observed. The relative discrimination capabilities of the techniques were similar. Penner serotyping fulfils more of the requisite criteria for typing methods.
Evaluation of Methods for SubtypingCampylobacter jejuniduring an Outbreak Involving a Food Handler
Authors: Fitzgerald Collette , Helsel Leta O. , Nicholson Mabel A. , Olsen Sonja J. , Swerdlow David L. , Flahart Robert , Sexton June , Fields Patricia I.
Published: July 2002

In October 1998, the Centers for Disease Control and Prevention (CDC) assisted in an investigation of an outbreak of campylobacteriosis at a school in Salina, Kansas. Twenty-two isolates were submitted from the Kansas state public health laboratory to CDC, 9 associated with the outbreak and 13 epidemiologically unrelated sporadic isolates. Pulsed-field gel electrophoresis (PFGE) usingSmaI andSalI was initially used to validate the epidemiologic data. We then tested the ability of other subtyping techniques to distinguish the outbreak-associated isolates from unrelated sporadic isolates. The methods employed were somatic O serotyping, PCR-restriction fragment length polymorphism (RFLP) analysis offlaA, DNA sequence analysis of 582 bp offlaAthat included the short variable region (SVR), and sequencing of the entireflaAgene. PFGE was the most discriminatory technique, yielding 11SmaI and 10SalI restriction profiles. All outbreak isolates were indistinguishable by PFGE, somatic O serotyping, and sequencing of the 582-bp region of theflaAgene.flatyping by PCR-RFLP grouped one sporadic isolate with the outbreak strain. Analysis of the DNA sequence of a 582-bp segment offlaAproduced strain groupings similar to that generated by PCR-RFLP but further differentiated twoflaAPCR-RFLP types (with a 1-bp difference in the 582-bp region). Two sporadic strains were distinct byflaAPCR-RFLP but differed only by a single base substitution in the 582-bp region. The entireflaAgene was sequenced from strains differing by a single base pair in the 582-bp region, and the data revealed that additional discrimination may in some cases be obtained by sequencing outside the SVR. PFGE was superior to all other typing methods tested for strain discrimination; it was crucial for understanding the Kansas outbreak and, whenSmaI was used, provided adequate discrimination between unrelated isolates.
Evaluation of the OptiMAL Rapid Antigen Test and Species-Specific PCR To Detect Placental
Plasmodium falciparum
Infection at Delivery
Authors: Mankhambo Limangeni , Kanjala Maxwell , Rudman Sarah , Lema Valentino M. , Rogerson Stephen J.
Published: July 2002

During pregnancy,
Plasmodium falciparum
infection of the placenta frequently occurs in the absence of parasites in peripheral blood. We investigated the abilities of the OptiMAL rapid immunochromatographic strip test for
P. falciparum
lactate dehydrogenase and species-specific PCR performed on peripheral blood to detect placental infection or malaria-associated low birth weight. Of 509 Malawian women screened by microscopy, 76 had malaria infection. Among these 509 women, the frequency of peripheral blood parasitemia was low. The OptiMAL test gave positive results in 37 of 171 women tested (one of whom had placental but not peripheral blood parasitemia) and had sensitivities of 71% for peripheral parasitemia and 38% for placental parasitemia compared to the microscopy values. The specificity for peripheral parasitemia was 94%. In 135 women, PCR had sensitivities of 94% for peripheral blood malaria detected by microscopy and 72% for placental infection. In samples examined by PCR, the prevalence of malaria in peripheral blood increased from 26.7% by microscopy to 51.9%. Women with placental malaria and women with malaria in peripheral blood samples by microscopy or OptiMAL testing, but not women with malaria detected only by PCR, had lower-birth-weight babies than did women without malaria by these criteria. Positive results by PCR in the absence of microscopic parasitemia were not associated with low birth weight. Neither OptiMAL nor PCR testing of peripheral blood is adequately sensitive to detect all placental malaria infection, but a positive result by OptiMAL testing identifies women with a high proportion of low-birth-weight babies.
Detection of Measles Virus-Specific Immunoglobulin M in Dried Venous Blood Samples by Using a Commercial Enzyme Immunoassay
Authors: Riddell Michaela A. , Leydon Jennie A. , Catton Mike G. , Kelly Heath A.
Published: July 2002

The optical densities (ODs) of 216 dried venous blood (DVB) samples submitted to the Victorian Infectious Diseases Reference Laboratory as part of enhanced measles surveillance were compared to the ODs of the corresponding serum samples collected at the same time. DVB samples, stored for up to 24 months at 4°C, were tested by the Dade Behring Enzygnost Anti-Measles-Virus/IgM immunoassay. Elution and testing conditions were optimized with the use of spiked DVB samples. The assay showed an overall sensitivity of 90.2% and a specificity of 98.8% for DVB samples compared to the results for serum. When the results were analyzed according to the length of time that the DVB sample had been stored, the assay was 100% sensitive and 97% specific according to the ODs for those samples stored for less than 6 months compared to the results for the corresponding serum samples, with 97.7% agreement between the results for the two sample types. These results demonstrate the potential for the use of DVB samples for the diagnosis of measles in routine diagnostic laboratories.
Successful Outcome of Treatment of a Disseminated Infection Due toFusarium dimerumin a Leukemia Patient
Authors: Letscher-Bru Valérie , Campos France , Waller Jocelyn , Randriamahazaka Raoul , Candolfi Ermanno , Herbrecht Raoul
Published: July 2002

We report the first case of proven disseminated infection due toFusarium dimerumassociated with a favorable outcome in a patient with acute leukemia and prolonged neutropenia. The patient presented persistent fever, multiple necrotic skin lesions, and bilateral pneumopathy.F. dimerumwas first isolated from three blood cultures and then from a skin biopsy and a mouth wash. Microscopy of positive blood cultures showed hyphae with phialides and few curved unicellular and some rare bicellular phialoconidia, permitting immediate presumptive identification of the genusFusarium. The patient failed to respond to conventional amphotericin B but recovered after treatment was switched to amphotericin B-lipid complex and neutrophil recovery.
Detection and Characterization of Rotaviruses in Hospitalized Neonates in Blantyre, Malawi
Authors: Cunliffe N. A. , Rogerson S. , Dove W. , Thindwa B. D. M. , Greensill J. , Kirkwood C. D. , Broadhead R. L. , Hart C. A.
Published: July 2002

In five separate fecal collections spanning three years, group A rotaviruses were detected by enzyme-linked immunosorbent assay in 35 (25%) of 142 specimens obtained from nondiarrheic, hospitalized neonates in Blantyre, Malawi. Molecular characterization of each strain identified, for the first time in neonates, a short electropherotype, genotype P[6], G8 strain type, similar to the dominant, cocirculating community strain detected in symptomatic infants in Blantyre. Partial sequence analysis of the VP4 and NSP4 genes of neonatal and community strains failed to identify changes which could explain the differences in clinical outcome. Neonatal serotype G8 rotaviruses should be considered as potential rotavirus vaccine candidates for use in Malawi.
A Hypothesis for DNA Viruses as the Origin of Eukaryotic Replication Proteins
Authors: Villarreal Luis P. , DeFilippis Victor R.
Published: July 2002

The eukaryotic replicative DNA polymerases are similar to those of large DNA viruses of eukaryotic and bacterial T4 phages but not to those of eubacteria. We develop and examine the hypothesis that DNA virus replication proteins gave rise to those of eukaryotes during evolution. We chose the DNA polymerase from phycodnavirus (which infects microalgae) as the basis of this analysis, as it represents a virus of a primitive eukaryote. We show that it has significant similarity with replicative DNA polymerases of eukaryotes and certain of their large DNA viruses. Sequence alignment confirms this similarity and establishes the presence of highly conserved domains in the polymerase amino terminus. Subsequent reconstruction of a phylogenetic tree indicates that these algal viral DNA polymerases are near the root of the clade containing all eukaryotic DNA polymerase delta members but that this clade does not contain the polymerases of other DNA viruses. We consider arguments for the polarity of this relationship and present the hypothesis that the replication genes of DNA viruses gave rise to those of eukaryotes and not the reverse direction.
Incoming Human Cytomegalovirus pp65 (UL83) Contained in Apoptotic Infected Fibroblasts Is Cross-Presented to CD8+T Cells by Dendritic Cells
Authors: Arrode Géraldine , Boccaccio Claire , Lulé Jacqueline , Allart Sophie , Moinard Nathalie , Abastado Jean-Pierre , Alam Antoine , Davrinche Christian
Published: July 2002

Human cytomegalovirus (HCMV) infection is well controlled mainly by cytotoxic CD8+T lymphocytes (CTL) directed against the matrix protein pp65 despite the numerous immune escape mechanisms developed by the virus. Dendritic cells (DCs) are key antigen-presenting cells for the generation of an immune response which have the capacity to acquire antigens via endocytosis of apoptotic cells and thus present peptides to major histocompatibility complex class I-restricted T cells. We examined whether this mechanism could contribute to the activation of anti-pp65 CTL. In this study, we show that infection by HCMV AD169 induced sensitization of MRC5 fibroblasts to tumor necrosis factor alpha-mediated apoptosis very early after virus inoculation and that pp65 contained in apoptotic cells came from the delivery of the matrix protein into the cell. We observed that immature DCs derived from peripheral monocytes were not permissive to HCMV AD169 infection but were able to internalize pp65-positive apoptotic infected MRC5 cells. We then demonstrated that following exposure to these apoptotic bodies, DCs could activate HLA-A2- or HLA-B35-restricted anti-pp65 CTL, suggesting that they acquired and processed properly fibroblast-derived pp65. Together, our data suggest that cross-presentation of incoming pp65 contained in apoptotic cells may provide a quick and efficient way to prime anti-HCMV CD8+T cells.
Adenovirus Types 11p and 35p Show High Binding Efficiencies for Committed Hematopoietic Cell Lines and Are Infective to These Cell Lines
Authors: Segerman Anna , Mei Ya-Fang , Wadell Göran
Published: July 2002

Hematopoietic cells are attractive targets for gene therapy. However, no satisfactory vectors are currently available. A major problem with the most commonly used adenovirus vectors, based on adenovirus type 2 (Ad2) or Ad5, is their low binding efficiency for hematopoietic cells. In this study we identify two adenovirus serotypes with high affinity for hematopoietic cells. The binding efficiency of prototype serotypes Ad4p, Ad11p, and Ad35p for different committed hematopoietic cell lines representing T cells (Jurkat), B cells (DG75), monocytes (U937-2), myeloblasts (K562), and granulocytes (HL-60) was evaluated and compared to that of Ad5v, the commonly used adenovirus vector, using flow cytometry. In contrast to Ad5v, which bound to less than 10% of the cells in all experiments, Ad11p and Ad35p showed high binding efficiency for all of the different hematopoietic cell lines. Ad4p bound to the lymphocytic cell lines to some extent but less well to the myelomonocytic cell lines. The abilities of the different serotypes to infect, replicate, and form complete infectious particles in the hematopoietic cell lines were also investigated by immunostaining,
35
S labeling of viral proteins, and titrations of cell lysates. Ad11p and Ad35p infected the highest proportion of cells, and Ad11p infected all of the cell lines investigated. The Ad11p hexon was expressed equally well in K562 and A549 cells. Jurkat cells also showed high levels of expression of Ad11p hexons, but the production of infectious particles was low. The binding properties of virions were correlated to their ability to infect and be expressed.
Viral Load Increases in SJL/J Mice Persistently Infected by Theiler's Virus after Inactivation of the β2mGene
Authors: Aubagnac Stéphanie , Brahic Michel , Bureau Jean-François
Published: July 2002

We show that inactivating the β2mgene increases the viral load of SJL/J mice persistently infected by Theiler's virus. Together with previous results, this shows that the characteristics ofTmevp1, a locus which controls the amount of viral RNA that persists in the central nervous system, are those of anH-2class I gene.
Molecular Basis for Attenuation of Neurovirulence of a Yellow Fever Virus/Japanese Encephalitis Virus Chimera Vaccine (ChimeriVax-JE)
Authors: Arroyo Juan , Guirakhoo Farshad , Fenner Sabine , Zhang Zhen-Xi , Monath Thomas P. , Chambers Thomas J.
Published: July 2002

A yellow fever virus (YFV)/Japanese encephalitis virus (JEV) chimera in which the structural proteins prM and E of YFV 17D are replaced with those of the JEV SA14-14-2 vaccine strain is under evaluation as a candidate vaccine against Japanese encephalitis. The chimera (YFV/JEV SA14-14-2, or ChimeriVax-JE) is less neurovirulent than is YFV 17D vaccine in mouse and nonhuman primate models (F. Guirakhoo et al., Virology 257:363–372, 1999; T. P. Monath et al., Vaccine 17:1869–1882, 1999). Attenuation depends on the presence of the JEV SA14-14-2 E protein, as shown by the high neurovirulence of an analogous YFV/JEV Nakayama chimera derived from the wild JEV Nakayama strain (T. J. Chambers, A. Nestorowicz, P. W. Mason, and C. M. Rice, J. Virol. 73:3095–3101, 1999). Ten amino acid differences exist between the E proteins of ChimeriVax-JE and the YFV/JEV Nakayama virus, four of which are predicted to be neurovirulence determinants based on various sequence comparisons. To identify residues that are involved in attenuation, a series of intratypic YFV/JEV chimeras containing either single or multiple amino acid substitutions were engineered and tested for mouse neurovirulence. Reversions in at least three distinct clusters were required to restore the neurovirulence typical of the YFV/JEV Nakayama virus. Different combinations of cluster-specific reversions could confer neurovirulence; however, residue 138 of the E protein (E
138
) exhibited a dominant effect. No single amino acid reversion produced a phenotype significantly different from that of the ChimeriVax-JE parent. Together with the known genetic stability of the virus during prolonged cell culture and mouse brain passage, these findings support the candidacy of this experimental vaccine as a novel live-attenuated viral vaccine against Japanese encephalitis.
Role of Hck in the Pathogenesis of Encephalomyocarditis Virus-Induced Diabetes in Mice
Authors: Choi K. S. , Jun H. S. , Kim H. N. , Park H. J. , Eom Y. W. , Noh H. L. , Kwon H. , Kim H. M. , Yoon J. W.
Published: July 2002

Soluble mediators such as interleukin-1β, tumor necrosis factor alpha (TNF-α), and inducible nitric oxide synthase (iNOS) produced from activated macrophages play an important role in the destruction of pancreatic β cells in mice infected with a low dose of the D variant of encephalomyocarditis (EMC-D) virus. The tyrosine kinase signaling pathway was shown to be involved in EMC-D virus-induced activation of macrophages. This investigation was initiated to determine whether the Src family of kinases plays a role in the activation of macrophages, subsequently resulting in the destruction of β cells, in mice infected with a low dose of EMC-D virus. We examined the activation of p59/p56Hck, p55Fgr, and p56/p53Lynin macrophages from DBA/2 mice infected with the virus. We found that p59/p56Hckshowed a marked increase in both autophosphorylation and kinase activity at 48 h after infection, whereas p55Fgrand p56/p53Lyndid not. The p59/p56Hckactivity was closely correlated with the tyrosine phosphorylation level of Vav. Treatment of EMC-D virus-infected mice with the Src kinase inhibitor, PP2, resulted in the inhibition of p59/p56Hckactivity and almost complete inhibition of the production of TNF-α and iNOS in macrophages and the subsequent prevention of diabetes in mice. On the basis of these observations, we conclude that the Src kinase, p59/p56Hck, plays an important role in the activation of macrophages and the subsequent production of TNF-α and nitric oxide, leading to the destruction of pancreatic β cells, which results in the development of diabetes in mice infected with a low dose of EMC-D virus.
Genetic Analysis of
cis
Regulatory Elements within the 5′ Region of the Human Papillomavirus Type 31 Upstream Regulatory Region during Different Stages of the Viral Life Cycle
Authors: Sen Ellora , Bromberg-White Jennifer L. , Meyers Craig
Published: July 2002

The function of the 5′ region of the upstream regulatory region (URR) in regulating E6/E7 expression in cancer-associated papillomaviruses has been largely uncharacterized. In this study we used linker-scanning mutational analysis to identify potential
cis
regulatory elements contained within a portion of the 5′ region of the URR that are involved in regulating transcription of the E6/E7 promoter at different stages of the viral life cycle. The mutational analysis illustrated differences in the transcriptional utilization of specific regions of the URR depending on the stage of the viral life cycle. This study identified (i) viral
cis
elements that regulate transcription in the presence and absence of any viral gene products or viral DNA replication, (ii) the role of host tissue differentiation in viral transcriptional regulation, and (iii)
cis
regulatory regions that are effected by induction of the protein kinase C pathway. Our studies have provided an extensive map of functional elements in the 5′ region (nuncleotides 7259 to 7510) of the human papillomavirus type 31 URR that are involved in the regulation of p99 promoter activity at different stages of the viral life cycle.
Regulation of DNA-Raised Immune Responses by Cotransfected Interferon Regulatory Factors
Authors: Sasaki Shin , Amara Rama Rao , Yeow Wen-Shuz , Pitha Paula M. , Robinson Harriet L.
Published: July 2002

Interferon regulatory factor 1 (IRF-1), IRF-3, and IRF-7 have been tested as genetic adjuvants for influenza virus hemagglutinin (HA) and nucleoprotein vaccine DNAs. Cotransfection of HA with IRF-3 and IRF-7 increased CD4 T-cell responses by 2- to 4-fold and CD8 T-cell responses by more than 10-fold. Following intramuscular deliveries of DNA, both CD4 and CD8 T cells were biased towards type 1 immune responses and the production of gamma interferon. Following gene gun bombardments of DNA, both were biased towards type 2 immune responses and the production of interleukin-4. The biases of the T-cell responses towards type 1 or type 2 were stronger for immunizations with IRF-3 as an adjuvant than for immunizations with IRF-7 as an adjuvant. Moderate adjuvant effects for antibody were observed. The isotypes of the antibody responses reflected the method of DNA delivery; intramuscular deliveries of DNA predominantly raised immunoglobulin G2a (IgG2a), whereas gene gun deliveries of DNA predominantly raised IgG1. These biases were enhanced by the codelivered IRFs. Overall, under the conditions of our experiments, IRF-3 had good activity for T cells, IRF-7 had good activity for both antibody and T cells, and IRF-1 had good activity for antibody.
Construction and Characterization of a Replication-Competent Retroviral Shuttle Vector Plasmid
Authors: Oh Jangsuk , Julias John G. , Ferris Andrea L. , Hughes Stephen H.
Published: July 2002

We constructed two versions of an RCASBP-based retroviral shuttle vector, RSVP (RCASBP shuttle vector plasmid), containing either the zeocin or blasticidin resistance gene. In this vector, the drug resistance gene is expressed in avian cells from the long terminal repeat (LTR) promoter, whereas in bacteria the resistance gene is expressed from a bacterial promoter. The vector contains a bacterial origin of replication (ColE1) to allow circular viral DNA to replicate as a plasmid in bacteria. The vector also contains the
lac
operator sequence, which binds to the
lac
repressor protein, providing a simple and rapid way to purify the vector DNA. The RSVP plasmid contains the following sequence starting with the 5" end: LTR,
gag
,
pol
,
env
, drug resistance gene,
lac
operator, ColE1, LTR. After this plasmid was transfected into DF-1 cells, we were able to rescue the circularized unintegrated viral DNA from RSVP simply by transforming the Hirt DNA into
Escherichia coli.
Furthermore, we were able to rescue the integrated provirus. DNA from infected cells was digested with an appropriate restriction enzyme (
Cla
I) and the vector-containing segments were enriched using
lac
repressor protein and then self-ligated. These enriched fractions were used to transform
E. coli
. The transformation was successful and we did recover integration sites, but higher-efficiency rescue was obtained with electroporation. The vector is relatively stable upon passage in avian cells. Southern blot analyses of genomic DNAs derived from successive viral passages under nonselective conditions showed that the cassette (drug resistance gene-
lac
operator-ColE1) insert was present in the vector up to the third viral passage for both resistance genes, which suggests that the RSVP vectors are stable for approximately three viral passages. Together, these results showed that RSVP vectors are useful tools for cloning unintegrated or integrated viral DNAs.
Differential Expression of Viral Bcl-2 Encoded by Kaposi's Sarcoma-Associated Herpesvirus and Human Bcl-2 in Primary Effusion Lymphoma Cells and Kaposi's Sarcoma Lesions
Authors: Widmer Isabelle , Wernli Marion , Bachmann Felix , Gudat Fred , Cathomas Gieri , Erb Peter
Published: July 2002

Expression of human herpesvirus 8 viral Bcl-2 protein was demonstrated in spindle cells of late-stage Kaposi's sarcoma lesions but not in primary effusion lymphoma cell lines. In contrast, strong expression of human Bcl-2 was found in stimulated primary effusion lymphoma cells, whereas in Kaposi's sarcoma lesions preferential mononuclear cells, and to a lesser extent spindle cells, stained positive.
Correlation of Equilibrium Data for Multicomponent Liquid-Liquid Systems
Authors: Sovilj Milan , Tolic Aleksandar , Maksimovic Zoran
Published: July 2002

Liquid liquid equilibrium studies have been carried out for five multicomponent systems comprising aromatics, paraffins and selected solvents (tetraethylene glycol, sulfolane, dimethyl sulfoxide, N-methylpyrrolidone and trimethyl phosphate). The experimental equilibrium data were fitted by empirical correlations suggested by Hand, Othmer-Tobias, Bulatov-Yachmenev and Rod. The best fit was obtained with the relation by Rod. The solvent capacities and selectivities are also compared.
Ceramic Foam in Catalytic Combustion of Methane
Authors: Jirátová Květa , Morávková Lenka , Malecha Jiří , Koutský Bohumil
Published: July 2002

Catalytic combustion of methane has been studied in the presence of catalysts supported on ceramic foams. Low concentrations of NOx (in the range of 5 - 50 mg m-3) were observed with all catalysts used. Concentrations of CO depended strongly on the type of the catalyst, its composition, methane throughputs and varied from 4 400 to 0 mg m-3. Minimum CO concentration (0 - 8 mg m-3) was gained with 0.04% Pt on ceramic foam modified with both alumina Al2O3 and lanthana La2O3.
Picosecond Tryptophan Fluorescence of Human Blood Serum Orosomucoid
Authors: Hof Martin , Vajda Stefan , Fidler Vlastimil , Karpenko Vladimír
Published: July 2002

The state of three tryptophyl residues in human serum orosomucoid was estimated by prediction methods based on parameters characterizing their hydrophobicity either directly, or in terms of buried surfaces of the individual amino acid residues. It is shown that tryptophan 25 is the most buried, while Trp 160 is the most exposed to the solvent. Trp 122 is in this respect in an intermediate state. The fluorescence decay behaviour was determined using a picosecond single photon counting system. The multiwavelength data were analyzed using a global analysis as well as a distribution of lifetimes program. Both procedures yielded the existence of four wavelength independent lifetimes (0.22 ns, 1.0 ns, 2.5 ns, and 8.4 ns). A tentative assignment of the decay associated spectra of the four components to the three individual tryptophans is presented.
On the Synthesis of 2-Substituted (η6-Indan-1,3-dione)tricarbonylchromium Complexes
Authors: Hrnčiar Peter , Hrnčiar Pavel , Gajda Vladimír , Švanygová Eva , Toma Štefan
Published: July 2002

Classical methods for the synthesis of indan-1,3-dione derivatives failed when applied to their tricarbonylchromium complexes. The desired 2-substituted (η6-indan-1,3-dione)tricarbonylchromium complexes were prepared starting from protected indan-1,3-dione derivatives such as bis(dioxolanes) and enol ethers. Tricarbonylchromium complexes of indan-1,3-dione having at least one hydrogen at C-2 are unstable. Nevertheless, ethylation of 2-methyl(η6-indan-1,3-dione)tricarbonylchromium and methylation of 2-ethyl(η6-indan-1,3-dione)tricarbonylchromium were possible and proceeded with high stereoselectivity.
Determination of the Charge Transfer Parameters of 4,4'-(Ferrocene-1,1'-diyldivinylene)bis(benzo-15-crown-5) at Glassy Carbon Electrode
Authors: Ghoneim Mohamed El-Metwally , Moharram Youssef Ibrahim , Taylor Norman
Published: July 2002

Electrochemical parameters of 4,4'-(ferrocene-1,1'-diyldivinylene)bis(benzo-15-crown-5) were collected using cyclic voltammetry, chronoamperometry and chronopotentiometry at a glassy carbon electrode. One-electron quasi-reversible oxidation wave was obtained at Epa of 0.532 V and Epc of 0.387 V. The peak separation, ∆Ep, standard heterogeneous rate constant, k0, and diffusion coefficient, D, of the electrochemical species have been processed by convolution and deconvolution transformations. Digital simulations using the kinetic parameters were also performed.
Stereoselective Synthesis of 2-Substituted Pyrrolidines
Authors: Deloisy Sandrine , Tietgen Heiko , Kunz Horst
Published: July 2002

Using O-pivaloyl protected D-galactopyranosylamine and D-arabinopyranosylamine, (S) or (R) configured α-substituted homoallylamines are synthesized with high diastereoselectivity by reaction of the corresponding aldimines with allyltributylstannane. Electrophile-induced endo-trig-cyclization of these N-glycosylhomoallylamines gave the 2-substituted pyrrolidines of high diastereomeric purity.
Simultaneous bilateral quadriceps tendon rupture while playing basketball
Authors: Shah M , Jooma N
Published: July 2002

Simultaneous bilateral quadriceps tendon rupture is an uncommon injury in healthy people and only a few cases have been reported in athletes. This is the first report of a patient with simultaneous bilateral quadriceps tendon rupture incurred while playing basketball. The injury was surgically repaired and the patient had a good functional outcome.
No evidence of persistent mumps virus infection in inflammatory bowel disease
Authors: Iizuka M , Saito H , Yukawa M , Itou H , Shirasaka T , Chiba M , Fukushima T , Watanabe S
Published: July 2002

BACKGROUND AND AIM

There is controversy regarding whether paramyxovirus infection is causally associated with inflammatory bowel disease (IBD). The latest cohort study claimed that atypical measles and mumps infections in childhood may be risk factors for later IBD. This study was conducted to clarify the validity of a causal link between persistent mumps virus infection and IBD.

SUBJECTS AND METHODS

(1) Amplification of the mumps virus genome was performed in both intestinal specimens (ulcerative colitis 15, Crohn's disease 15, control 10) and peripheral blood lymphocytes (PBL) (ulcerative colitis seven, Crohn's disease six, control three) by reverse transcription-polymerase chain reaction (RT-PCR) followed by Southern hybridisation using primers specific to the viral genome encoding phosphoprotein or haemagglutinin-neuraminidase. (2) Titre of serum antimumps IgG was measured in 16 patients with ulcerative colitis, in 16 patients with Crohn's disease, and in 16 normal controls using an enzyme linked immunosorbent assay.

RESULTS

(1) The mumps virus genome was not detected by RT-PCR in intestinal specimens or PBL in any case. (2) Antimumps IgG titre was positive in 7/16 ulcerative colitis, 10/16 Crohn's disease, and 11/16 control specimens. The mean (SEM) titre of antimumps IgG was 12.281 (7.831) in ulcerative colitis, 7.675 (1.608) in Crohn's disease, and 8.637 (1.969) in controls, with no significant difference between the three groups.

CONCLUSION

We could not find any evidence to support a causal link between persistent mumps virus infection and IBD.
Prevalence of mental illness in a rehabilitation unit for older adults
Authors: Shah D C , Evans M , King D
Published: July 2002

The prevalence of psychiatric disorders was studied in 78 elderly people in a rehabilitation unit for older adults. The patients were assessed using the Evans Liverpool depression rating scale, Hospital Anxiety and Depression scale and Mini-Mental State Examination. Twenty-eight (35.9%) patients were found to be depressed, 15 of these also had raised anxiety. Thirty-one (41.0%) patients had significant cognitive impairment and 14 of these had associated depression. Only 33 (42%) had no evidence of either cognitive impairment or mood disorder. On discharge, 20 (25.6%) patients were on antidepressant treatment but only 50% of those had this diagnosis recorded on the discharge summary. Our results showed higher prevalence of depression in this situation compared with the reported prevalence of 20–30% in the acute hospital setting. We recommend that all patients undergoing rehabilitation should be routinely screened for depression as it is common and treatment will improve the overall outcome.
Causes of anorexia in untreated hyperthyroidism: a prospective study
Authors: Dai Wei-Xin , Meng Xun-Wu
Published: July 2002

Seventeen consecutive patients (mean (SD) 46 (11) years) with untreated hyperthyroidism and anorexia and 29 patients (35 (9) years) with untreated hyperthyroidism without anorexia were studied. The study was conducted at the thyroid clinic of the PUMC Hospital, Beijing, China from March to August 1997. The patients' ages, serum free calcium, liver function and emotional state, specifically the level of anxiety (using the self anxiety scale, Chinese version), were compared before and/or after antithyroid drug treatment in the two groups. This prospective study suggested that the causes of anorexia in untreated hyperthyroidism are complicated. Older age, abnormal liver function, and the level of anxiety are significantly related to anorexia in untreated hyperthyroidism, but hypercalcaemia was not confirmed to be related to anorexia in the study.
Tobacco sponsorship of Formula One and CART auto racing: tobacco brand exposure and enhanced symbolic imagery through co-sponsors' third party advertising
Authors: Dewhirst T , Hunter A
Published: July 2002

Through third party advertising of events such as Formula One and CART auto racing, tobacco brand names continue to attain visibility to a vast audience
A molecularmorphological approach solves taxonomic controversy in arcticDraba(Brassicaceae)
Authors: Scheen Anne-Cathrine , Elven Reidar , Brochmann Christian
Published: July 2002

We used a combined molecular and morphological approach to solve a long-term controversy in the taxonomically complex genus Draba L. (Brassicaceae): the delimitation of Draba lactea Adams versus Draba fladnizensis Wulfen. We also tested hypotheses on the origin of D. lactea, which has been reported as hexaploid and possibly derived from the diploids D. fladnizensis, Draba nivalis Liljeblad, and (or) Draba subcapitata Simmons. In an initial analysis of large population samples of three of these autogamous species, intrapopulational isozyme variation was low or absent (mean genotypic diversity, D, was 0.10), the diploids were almost invariably homozygous, and D. lactea was highly fixed-heterozygous. In the main analysis, we examined 3547 populations of the four tentative species from the arctic archipelago of Svalbard for variation in isozymes, random amplified polymorphic DNAs (RAPDs), and 36 morphological characters. Multivariate analyses of the RAPD data revealed four very distinct groups of multilocus phenotypes. These groups also differed in several morphological characters and corresponded to the four tentative species. The species were less differentiated at isozyme loci, in particular the diploids D. fladnizensis and D. subcapitata, but D. lactea was clearly distinguished from D. fladnizensis based on all three data sets. Contrary to most earlier suggestions, the hexaploid D. lactea was genetically more similar to D. subcapitata than to D. nivalis, but our analyses of the material from Svalbard did not give conclusive evidence on the origin of this widespread arctic hexaploid.Key words: arctic, Draba, isozymes, morphology, polyploidy, RAPDs.
Effets de quelques pratiques horticoles de conduite des plantes sur la production de roses coupées
Authors: Gudin Serge , Coulon Anne , Bris Manuel Le
Published: July 2002

Four different plant management treatments, corresponding with different existing growing practices, were compared for flower production of Rosa hybrida L. cv. Meiqualis plants grown for 1 year in a greenhouse in the south of France. Two of them used shoot bending during plant formation and subsequent production management. The harvest level practiced on the thickest stems differentiated these two treatments. Two other treatments consisted in using shoot removal and pruning during plant formation. In the first one, during production management, unmarketable shoots were removed by pinching, whereas they were bent and left on the plant in the second one. This last treatment resulted in the highest yield of marketable stems and stem quality after 1 year, although the treatment using shoot bending during plant formation and production associated with a high harvest level on the thickest stems produced the largest quantity during autumn and winter. The results are discussed with reference to different yield components determined by the different plant management treatments applied, such as inter-flush cycle time, bud sprouting, occurrence of certain types of unmarketable stems (blind shoots and bowed peduncled stems), and development of new bottom breaks.Key words: shoot bending, bottom breaks, blind shoots, bowed stems, production, Rosa hybrida L.
A stage-explicit expression of the von Bertalanffy growth model for understanding age at first reproduction of Great Lakes fishes
Authors: He Ji X , Stewart Donald J
Published: July 2002

An initial annual growth rate of body length and its regular decrease with increasing age has general linkages with age at first reproduction (tR). We clarify their combinations and develop predictive functions. We use a complete FordWalford plot with yearling size (L1) on the y axis and show a slope transition between the relative yearling growth rate (ρy) and the FordWalford slope (ρ). The three stage-specific variables define a complete body-length trajectory over ages, including all von Bertalanffy growth parameters and the FordWalford intercept (Lint). The difference between asymptotic length (Linf) and yearling length is growth potential after the first annulus. Yearling growth is a transition period, so growth potential can be adjusted as ρLinf or Linf  Lint. Changes in the three life-stage variables have contrasting effects on growth potential and von Bertalanffy growth parameters, so they have contrasting relations with tR. For most invertebrate-eating fishes in the Laurentian Great Lakes, dominant changes in growth trajectories were reflected in ρ, so tR was predicted by the von Bertalanffy growth coefficient, K. For walleye (Stizostedion vitreum) populations around the Great Lakes, dominant changes in growth trajectories were from yearling size or yearling growth, so tR was predicted using Lint. Our results have clear implications for understanding fish population dynamics.
Expressed sequence tags for the chicken genome from a normalized 10-day-old white leghorn whole-embryo cDNA library. 3. DNA sequence analysis of genetic variation in commercial chicken populations
Authors: Smith E J , Shi L , Smith G
Published: July 2002

Single nucleotide polymorphisms (SNPs) have emerged as a major class of DNA markers with the advantage of permitting the development of high-density genetic maps adequate for quantitative trait loci (QTL) identification by linkage-disequilibrium analysis. Here we describe results of a relatively high-depth survey of chicken broiler and layer populations for SNPs in targeted genomic regions of chicken expressed sequence tag (EST) sites. The sequences scanned, representing the composite sequence of 12 amplified fragments for a total of 6489 bp, were randomly distributed, occurring on six different chromosomes or linkage groups in the chicken genome. Although one of the genomic DNA sequences did not match the reference cDNA sequence, another contained an intron that separated two putative exons. The number of SNPs observed within each of the 12 EST-targeted genomic regions ranged from 0 to 10 for a total of 44 and a frequency of 0.7%. About 70% of the polymorphisms were shared between layer and broiler populations. The average heterozygosity within the populations ranged from 0.15 to 0.48, with the layer populations showing the higher heterozygosity. SNPs and oligonucleotides described will provide a resource for genetic analysis in commercial chicken populations. The data appear to indicate that the relative frequency of SNPs in the targeted regions scanned is higher than the frequency reported for any of the other regions scanned to date in other eukaryotic genomes. Additionally, the results suggest that the use of DNA pools may offer an efficient approach to SNP detection in chickens, as has been shown in other vertebrates.Key words: DNA sequence analysis, SNPs, expressed sequence tags, chicken.