bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peritoneal Dialysis-Associated Peritonitis Caused by
Dermabacter hominis
Authors: Radtke Andreas , Bergh Kåre , Øien Cecilia M. , Bevanger Lars S. 
Published: July 2002

ABSTRACT

Dermabacter hominis
was the cause of a peritoneal dialysis-associated peritonitis.
D. hominis
was identified by phenotypic criteria and by sequencing the 16S rRNA gene. Clinical cure was achieved with cefuroxime treatment despite the isolate's reduced susceptibility to this drug (MIC, 12 mg/liter) on in vitro testing. The successful treatment was probably due to the high concentrations attained by intraperitoneal administration of the drug.
Evaluation of a Rapid PCR-Based Epidemiological Typing Method for Routine Studies of
Mycobacterium tuberculosis
Authors: Yates Malcolm D. , Drobniewski Francis A. , Wilson Stuart M. 
Published: July 2002

ABSTRACT

Restriction fragment length polymorphism (RFLP) based on the insertion sequence IS
6110
is used to investigate episodes of suspected transmission of infection of tuberculosis but usually takes a number of weeks from receipt of request to obtain a result. Often investigations would benefit from a more rapid method, possibly one containing an amplification step. The method employed uses a simple DNA extraction followed by a PCR step involving a single primer. Restriction enzyme analysis was performed when the patterns obtained from the PCR products were indistinguishable, especially when only single similar-size bands were obtained. The isolates used were strains of
Mycobacterium tuberculosis
submitted for epidemiological investigations as part of (i) possible contact-outbreak (22 episodes involving between 2 and 20 patients), (ii) possible incidents of laboratory cross-contamination (21 episodes), and (iii) possible change in drug resistance pattern or a case of reinfection (1 patient). The PCR products giving similar patterns were then subjected to restriction enzyme analysis. In conclusion it has been shown that this method is rapid, with results within 1 to 2 days of the request being received; is reproducible; and gives the same results as does RFLP. The restriction enzyme analysis stage has improved the efficiency of the technique.
New Simple and Rapid Test for Culture Confirmation of
Mycobacterium tuberculosis
Complex: a Multicenter Study
Authors: Hasegawa Naoki , Miura Takao , Ishii Koudou , Yamaguchi Kazuhiro , Lindner Thomas H. , Merritt Samuel , Matthews Janis D. , Siddiqi Salman H. 
Published: July 2002

ABSTRACT

Mycobacterial antigen MPB64 has been identified as a
Mycobacterium tuberculoisis
complex-specific secretory protein since 1984. Recently, a simple culture confirmation test for
M. tuberculosis
complex has been developed by using lateral flow immunochromatographic assay (ICA) to detect MPB64 with anti-MPB64 monoclonal antibody. The current multicenter study evaluated the performance of an ICA slide test for MPB64 antigen in the clinical setting. Primary positive cultures from clinical specimens, as well as stock cultures, were tested. Approximately 100 μl of positive liquid culture medium or suspension made from colonies on solid medium was placed into the test well of the plastic slide devise, and the test was read after 15 min. No processing or instrumentation was required. A total of 304 mycobacterial isolates consisting of
M. tuberculosis
complex (171 isolates) and mycobacteria other than
M. tuberculosis
(MOTT) complex (133 isolates) belonging to 18 different species were tested. Growth in liquid media (Mycobacteria Growth Indicator Tube [MGIT] and Radiometric 12B), as well as in solid (Löwenstein-Jensen and Middlebrook 7H10 agar) media, was evaluated. Results were compared with those obtained with nucleic acid-based and/or high-pressure liquid chromatography identification. All MOTT were found to be negative on the ICA slide with no cross-reaction. All
M. tuberculosis
and
M. africanum
cultures were found to be positive, whereas the results of
M. bovis
and
M. bovis
BCG cultures were variable since some of the BCG strains are known to lack MPB64 antigen production. The results did not change with prolonged storage of cultures. This low-tech rapid test with high sensitivity and specificity could provide an alternative to currently available identification methods, particularly for recently introduced nonradiometric liquid culture systems such as MGIT.
Ribosomal DNA-Directed PCR for Identification of
Achromobacter
(
Alcaligenes
)
xylosoxidans
Recovered from Sputum Samples from Cystic Fibrosis Patients
Authors: Liu Lixia , Coenye Tom , Burns Jane L. , Whitby Paul W. , Stull Terrence L. , LiPuma John J. 
Published: July 2002

ABSTRACT

The opportunistic human pathogen
Achromobacter
(
Alcaligenes
)
xylosoxidans
has been recovered with increasing frequency from respiratory tract culture of persons with cystic fibrosis (CF). However, confusion of this species with other closely related respiratory pathogens has limited studies to better elucidate its epidemiology, natural history, and pathogenic role in CF. Misidentification of
A. xylosoxidans
as
Burkholderia cepacia
complex is especially problematic and presents a challenge to effective infection control in CF. To address the problem of accurate identification of
A. xylosoxidans
, we developed a PCR assay based on a 16S ribosomal DNA sequence. In an analysis of 149 isolates that included 47
A. xylosoxidans
and several related glucose-nonfermenting species recovered from CF sputum, the sensitivity and specificity of this PCR assay were determined to be 100 and 97%, respectively. The availability of this assay will enhance identification of
A. xylosoxidans
, thereby facilitating study of the pathogenic role of this species and improving infection control efforts in CF.
Simultaneous Differentiation and Typing of
Entamoeba histolytica
and
Entamoeba dispar
Authors: Zaki Mehreen , Meelu Parool , Sun Wei , Clark C. Graham 
Published: July 2002

ABSTRACT

Sequences corresponding to some of the polymorphic loci previously reported from
Entamoeba histolytica
have been detected in
Entamoeba dispar
. Comparison of nucleotide sequences of two loci between
E. dispar
strain SAW760 and
E. histolytica
strain HM-1:IMSS revealed significant differences in both repeat and flanking regions. The tandem repeat units varied not only in sequence but also in number and arrangement between the two species at both the loci. Using the sequences obtained, primer pairs aimed at amplifying species-specific products were designed and tested on a variety of
E. histolytica
and
E. dispar
samples. Amplification results were in complete agreement with the original species classification in all cases, and the PCR products displayed discernible size and pattern variations among the isolates.
Evaluation of Eight Rapid Screening Tests for Acute Leptospirosis in Hawaii
Authors: Effler Paul V. , Bogard April K. , Domen Harry Y. , Katz Alan R. , Higa Henry Y. , Sasaki David M. 
Published: July 2002

ABSTRACT
Leptospirosis is a major public health problem throughout the world. Clinical recognition of leptospirosis is challenging, and the definitive serologic diagnostic assay, the microscopic agglutination test, is time-consuming and difficult to conduct. Various serologic screening tests have been developed, but their performance among ill persons in the United States has not been established. Eight screening tests were compared using 379 serum samples obtained in 1998 and 1999 from a series of 236 patients (33 with confirmed infection). The median number of days between illness onset and specimen collection was 9. The overall sensitivity, by specimen, for each test was as follows: indirect hemagglutination assay (MRL Diagnostics, Cypress, Calif.), 29%; INDX Leptospira Dip-S-Tick (PanBio InDx, Inc., Baltimore, Md.), 52%; Biognost IgM IFA test (Bios GmbH Labordiagnostik, Gräfelfing, Germany), 40%; Biolisa IgM ELISA (Bios GmbH, Labordiagnostik), 48%; Leptospira IgM ELISA (PanBio Pty Ltd., Brisbane, Australia), 36%; SERION ELISA classic Leptospira (Institut Virion•Serion GmbH, Würzburg, Germany), 48%; LEPTO Dipstick(Organon-Teknika, Ltd., Amsterdam, The Netherlands), 34%; Biosave latex agglutination test (LATEX; Bios GmbH Labordiagnostik), 86%. Test specificity ranged from 85 to 100% among all tests except LATEX, for which the specificity was significantly lower, at 10%. Test sensitivity was particularly low (<25%) for all tests (except LATEX) on specimens collected during the first week of illness. This is the most comprehensive field trial of leptospirosis screening tests reported to date. The data indicate that immunoglobulin M detection tests have limited utility for diagnosing leptospirosis during the initial evaluation of patients seen in Hawaii, a time when important therapeutic decisions are made. Improved leptospirosis screening tests are needed.
AMF-1/Gps2 Binds p300 and Enhances Its Interaction with Papillomavirus E2 Proteins
Authors: Peng Yu-Cai , Breiding David E. , Sverdrup Francis , Richard James , Androphy Elliot J. 
Published: July 2002

ABSTRACT
The cellular protein AMF-1 (Gps2) positively modulates gene expression by the papillomavirus E2 protein (D. E. Breiding et al., Mol. Cell. Biol. 17:7208–7219, 1997). We show here that AMF-1 also binds the transcriptional coactivator p300 in vitro and in vivo. E2 interacted weakly with p300. These observations led to a model in which AMF-1 recruits p300 into a complex with E2. Cotransfection of AMF-1 or p300 stimulated levels of E2-dependent transcription, while cotransfection of both AMF-1 and p300 showed an additive effect. The functional significance of p300 recruitment for E2 transactivation was evidenced by repression of E2-activated transcription by adenovirus E1A, which inhibits both coactivator and acetylase activities of p300. Antibodies to AMF-1 or E2 immunoprecipitated histone acetylase activity from cell lysates. Western blotting using antibody against acetyl-lysine failed to detect acetylation of AMF-1 or E2 in complex with p300. These results suggest that AMF-1 facilitates the recruitment of p300 and its histone acetylase activity into complexes with E2 and represents a novel mechanism of transcriptional activation.
Respiratory Syncytial Virus G and/or SH Glycoproteins Modify CC and CXC Chemokine mRNA Expression in the BALB/c Mouse
Authors: Tripp Ralph A. , Jones Les , Anderson Larry J. 
Published: July 2002

ABSTRACTChemokine mRNA expression by pulmonary leukocytes following infection of BALB/c mice with two strains of respiratory syncytial virus (RSV) and one strain of parainfluenza virus type 3 (PIV-3) was determined. The results suggest that RSV G and/or SH proteins inhibit early MIP-1α, MIP-1β, MIP-2, MCP-1, and IP-10 mRNA expression. TCA-3 mRNA expression was found to be increased during PIV-3 infection.
Novel Mouse Type D Endogenous Proviruses and ETn Elements Share Long Terminal Repeat and Internal Sequences
Authors: Mager Dixie L. , Freeman J. Douglas 
Published: July 2002

ABSTRACTThe repetitive ETn (early transposon) family of sequences represents an active “mobile mutagen” in the mouse genome. The presence of long terminal repeats (LTRs) and other diagnostic features indicate that ETns are retrotransposons but they contain no long open reading frames or documented similarity to the genes of known retroviruses or other retroelements. Thus, the mechanisms responsible for the mobility of this family have been unknown. In this study, we used computer searches to detect a small region of previously unrecognized type D retroviralpolhomology within ETn elements. This small region was used to isolate two mouse endogenous proviral elements withgag,pro, andpolgenes similar to simian type D viruses. This new family of mouse endogenous proviruses, termed MusD, is present in several hundred copies in the genome. Interestingly, the MusD LTRs, 3′ internal region, and the 5′ region expected to contain the packaging signal are very closely related to members of the ETn subfamily that have recently transposed. Analysis of different mouse strains indicates that MusD elements predate the existence of the mobile subfamily of ETns. These findings indicate that the ETn family was likely created via recombination events resulting in a near complete substitution of MusD coding sequences with unrelated DNA. Furthermore, these results suggest that ETn transcripts retrotranspose using proteins provided by MusD proviruses.
Processing of α-Globin and ICP0 mRNA in Cells Infected with Herpes Simplex Virus Type 1 ICP27 Mutants
Authors: Ellison Kimberly S. , Rice Stephen A. , Verity Robert , Smiley James R. 
Published: July 2002

ABSTRACT

Herpes simplex virus (HSV) ICP27 is an essential and multifunctional regulator of viral gene expression that modulates RNA splicing, polyadenylation, and nuclear export. We have previously reported that ICP27 causes the cytoplasmic accumulation of unspliced α-globin pre-mRNA. Here we examined the effects of a series of ICP27 mutations that alter important functional regions of the protein on the processing and nuclear transport of α-globin and HSV ICP0 RNA. The results demonstrate that ICP27 mutants that are impaired for growth in noncomplementing cells, including mutants in the N- and C-terminal regions, are defective in the accumulation of α-globin pre-mRNA. Unexpectedly, several mutants that are competent to repress the expression of reporter genes in transient transfection assays failed to accumulate unspliced RNA, implying that different mechanisms are responsible for transrepression and pre-mRNA accumulation. Several mutants caused a marked increase in the length and heterogeneity of the α-globin mRNA poly(A) tail, suggesting that ICP27 may directly or indirectly affect the regulation of poly(A) polymerase. ICP27 was also required for the accumulation of multiple ICP0 intron-bearing transcripts, but this effect displayed a mutational sensitivity profile different from that of accumulation of unspliced α-globin RNA. Moreover, unlike spliced and unspliced α-globin RNAs, which were efficiently exported to the cytoplasm, spliced and intron-containing ICP0 transcripts were predominantly nuclear in localization, and ICP27 was not required for nuclear retention of the spliced message. We propose that these transcript- and ICP27 allele-specific differences may be explained by the presence of a strong
cis
-acting ICP27 response element in the α-globin transcript.
Upregulation of Tyrosine Kinase TKT by the Epstein-Barr Virus Transactivator Zta
Authors: Lu Jean , Chen Shao-Yin , Chua Huey-Huey , Liu Yu-Sheng , Huang Yu-Tzu , Chang Yao , Chen Jen-Yang , Sheen Tzung-Shiahn , Tsai Ching-Hwa 
Published: July 2002

ABSTRACTThe Zta protein is a key transactivator involved in initiating the Epstein-Barr virus (EBV) lytic cascade. In addition to transactivating many viral genes, Zta has the capacity to influence host cellular signals by binding to promoter regions or by interacting with several important cellular factors. Based on the observation that tyrosine kinases play central roles in determining the fate of cells, a kinase display assay was used to investigate whether cells expressing Zta have an altered pattern of kinase expression. The assay revealed that TRK-related tyrosine kinase (TKT) is expressed at significant levels in Zta transfectants but not in control cells. Additional evidence was obtained from Northern and Western blotting. Importantly, the upregulation of phosphorylated TKT and TKT downstream effector matrix metalloproteinase 1 in Zta transfectants hinted that TKT might initiate a signaling cascade in Zta-expressing cells. In addition, deletion analysis of the Zta protein revealed that the transactivation and dimerization domains were both essential for the upregulation of TKT transcription. Moreover, correlation of expression levels of Zta and TKT transcripts in nasopharyngeal carcinoma biopsy specimens was clearly demonstrated by quantitative PCR (Q-PCR), which provides the first evidence for an effect of Zta on cellular gene expression in vivo. These findings offer insight into the virus-cell interactions and may help us elucidate the role of EBV in tumorigenesis.
Differential Dynamics of CD4+and CD8+T-Lymphocyte Proliferation and Activation in Acute Simian Immunodeficiency Virus Infection
Authors: Kaur Amitinder , Hale Corrina L. , Ramanujan Saroja , Jain Rakesh K. , Johnson R. Paul 
Published: July 2002

ABSTRACTAlthough lymphocyte turnover in chronic human immunodeficiency virus and simian immunodeficiency virus (SIV) infection has been extensively studied, there is little information on turnover in acute infection. We carried out a prospective kinetic analysis of lymphocyte proliferation in 13 rhesus macaques inoculated with pathogenic SIV. A short-lived dramatic increase in circulating Ki-67+lymphocytes observed at 1 to 4 weeks was temporally related to the onset of SIV replication. A 5- to 10-fold increase in Ki-67+CD8+T lymphocytes and a 2- to 3-fold increase in Ki-67+CD3−CD8+natural killer cells accounted for >85% of proliferating lymphocytes at peak proliferation. In contrast, there was little change in the percentage of Ki-67+CD4+T lymphocytes during acute infection, although transient increases in Ki-67−and Ki-67+CD4+T lymphocytes expressing CD69, Fas, and HLA-DR were observed. A two- to fourfold decline in CD4+T lymphocytes expressing CD25 and CD69 was seen later in SIV infection. The majority of Ki-67+CD8+T lymphocytes were phenotypically CD45RA−CD49dhiFashiCD25−CD69−CD28−HLA-DR−and persisted at levels twofold above baseline 6 months after SIV infection. Increased CD8+T-lymphocyte proliferation was associated with cell expansion, paralleled the onset of SIV-specific cytotoxic T-lymphocyte activity, and had an oligoclonal component. Thus, divergent patterns of proliferation and activation are exhibited by CD4+and CD8+T lymphocytes in early SIV infection and may determine how these cells are differentially affected in AIDS.
Helicase and Capping Enzyme Active Site Mutations in Brome Mosaic Virus Protein 1a Cause Defects in Template Recruitment, Negative-Strand RNA Synthesis, and Viral RNA Capping
Authors: Ahola Tero , den Boon Johan A. , Ahlquist Paul 
Published: July 2002

ABSTRACT

Brome mosaic virus (BMV) encodes two RNA replication proteins: 1a, which contains RNA capping and helicase-like domains, and 2a, which is related to polymerases. BMV 1a and 2a can direct virus-specific RNA replication in the yeast
Saccharomyces cerevisiae
, which reproduces the known features of BMV replication in plant cells. We constructed single amino acid point mutations at the predicted capping and helicase active sites of 1a and analyzed their effects on BMV RNA3 replication in yeast. The helicase mutants showed no function in any assays used: they were strongly defective in template recruitment for RNA replication, as measured by 1a-induced stabilization of RNA3, and they synthesized no detectable negative-strand or subgenomic RNA. Capping domain mutants divided into two groups. The first exhibited increased template recruitment but nevertheless allowed only low levels of negative-strand and subgenomic mRNA synthesis. The second was strongly defective in template recruitment, made very low levels of negative strands, and made no detectable subgenomes. To distinguish between RNA synthesis and capping defects, we deleted chromosomal gene
XRN1
, encoding the major exonuclease that degrades uncapped mRNAs.
XRN1
deletion suppressed the second but not the first group of capping mutants, allowing synthesis and accumulation of large amounts of uncapped subgenomic mRNAs, thus providing direct evidence for the importance of the viral RNA capping function. The helicase and capping enzyme mutants showed no complementation. Instead, at high levels of expression, a helicase mutant dominantly interfered with the function of the wild-type protein. These results are discussed in relation to the interconnected functions required for different steps of positive-strand RNA virus replication.
Generation of Mutant Murine Cytomegalovirus Strains from Overlapping Cosmid and Plasmid Clones
Authors: Ehsani Mariam E. , Abraha Tshge W. , Netherland-Snell Cecile , Mueller Niklaus , Taylor Meghan M. , Holwerda Barry 
Published: July 2002

ABSTRACT
We have developed a cosmid and plasmid system to generate mutant strains of murine cytomegalovirus (MCMV). The system is based on a series of seven overlapping cosmid clones that regenerate MCMV when cotransfected into mouse cells. The unaltered cosmids produce MCMV that is indistinguishable from wild-type MCMV based on restriction enzyme digest patterns of virus DNA and growth rates both in vitro and in vivo. Analysis of viral DNA from plaque-purified recombinant isolates taken from in vitro and in vivo stocks indicated that regeneration did not introduce novel mutations in the recombinant viral genomes. Isolation of specific genes and subsequent generation of specific mutant MCMVs was accomplished by replacement of cosmids with overlapping plasmid subclones. A new vector, PmeSUB, featuring a multiple cloning site and a stringent origin of replication, was constructed to make large subclones for use with smaller subclones containing the gene of interest. The utility of this system was demonstrated by the generation of two different mutant MCMVs from different combinations of overlapping plasmid subclones of one cosmid. The advantages of this system are that (i) target genes are maintained as small clones making them amenable to standard in vitro mutagenesis manipulations and that (ii) no reporter or selection genes are necessary to identify mutants.
Direct Inhibitory Effect of Rotavirus NSP4(114-135) Peptide on the Na
+
-
d
-Glucose Symporter of Rabbit Intestinal Brush Border Membrane
Authors: Halaihel Nabil , Liévin Vanessa , Ball Judith M. , Estes Mary K. , Alvarado Francisco , Vasseur Monique 
Published: July 2002

ABSTRACT

The direct effect of a rotavirus nonstructural glycoprotein, NSP4, and certain related peptides on the sodium-coupled transport of
d
-glucose and of
l
-leucine was studied by using intestinal brush border membrane vesicles isolated from young rabbits. Kinetic analyses revealed that the NSP4(114-135) peptide, which causes diarrhea in young rodents, is a specific, fully noncompetitive inhibitor of the Na
+
-
d
-glucose symporter (SGLT1). This interaction involves three peptide-binding sites per carrier unit. In contrast, the Norwalk virus NV(464-483) and mNSP4(131K) peptides, neither of which causes diarrhea, both behave inertly. The NSP4(114-135) and NV(464-483) peptides inhibited Na
+
-
l
-leucine symport about equally and partially via a different transport mechanism, in that Na
+
behaves as a nonobligatory activator. The selective and strong inhibition caused by the NSP4(114-135) peptide on SGLT1 in vitro suggests that during rotavirus infection in vivo, NSP4 can be one effector directly causing SGLT1 inhibition. This effect, implying a concomitant inhibition of water reabsorption, is postulated to play a mechanistic role in the pathogenesis of rotavirus diarrhea.