bluuebunny/crossref_metadata_2025_split huggingface dataset:
Nationwide German Multicenter Study on Prevalence of Antibiotic Resistance in Staphylococcal Bloodstream Isolates and Comparative In Vitro Activities of Quinupristin-Dalfopristin
Authors: von Eiff Christof , Reinert Ralf René , Kresken Michael , Brauers Johannes , Hafner Dieter , Peters Georg 
Published: December 2019

ABSTRACT

Antibiotic-resistant gram-positive bacteria have become an increasing problem in the last two decades. In order to evaluate the prevalence of antibiotic resistance in staphylococcal bloodstream isolates in Germany, 2,042 staphylococci collected in 21 tertiary-care hospitals were investigated during a 3-year period (March 1996 to March 1999). Altogether, 1,448
S. aureus
isolates and 594 coagulase-negative staphylococci (CoNS) that comprised 13 different species were included. Furthermore, the antistaphylococcal activities of quinupristin-dalfopristin were compared with those of eight other compounds by the broth microdilution method. The rates of oxacillin resistance in
Staphylococcus aureus
,
S. epidermidis
,
S. haemolyticus
, and other CoNS were 13.5, 69, 90, and 34%, respectively. In oxacillin-resistant strains high rates of resistance (up to 100%) to erythromycin, clindamycin, ciprofloxacin, and gentamicin were also observed. However, no strain appeared to be resistant to vancomycin or quinupristin-dalfopristin. The streptogramin combination exhibited excellent in vitro activity against all staphylococcal species tested, regardless of the patterns of resistance to other drug classes. In terms of MICs at which 90% of the isolates are inhibited, quinupristin-dalfopristin was 2 times more active against
S. aureus
isolates, 4 to 16 times more active against
S. haemolyticus
, and 8 to 32 times more active against
S. epidermidis
than vancomycin or teicoplanin.
Comparison of an Established Antibody Sandwich Method with an Inhibition Method of
Histoplasma capsulatum
Antigen Detection
Authors: Garringer Todd O. , Wheat L. Joseph , Brizendine Edward J. 
Published: December 2019

ABSTRACT

The
Histoplasma
antigen immunoassay utilizes an antibody sandwich method that provides a rapid and reliable means of diagnosing the more severe forms of histoplasmosis. Inhibition assays have been developed for antigen detection and offer at least one potential advantage, namely, reduced antibody requirements. We have developed an inhibition assay using the polyclonal antibody employed in our standard sandwich assay. Urine and serum specimens from patients with culture-proven histoplasmosis and controls were tested using both methods. The two methods had similar sensitivities for detection of antigen in urine (antibody sandwich = 92.5% versus inhibition = 87.5%,
P
= 0.500) and serum (82.5% versus 80.0%,
P
= 1.000). With serum, the specificities of both methods were similar (antibody sandwich assay = 95.0% versus inhibition assay = 92.5%,
P
= 1.000), and with urine, the specificity of the antibody sandwich method was superior (97.5% versus 80.0%,
P
= 0.039). While the overall reproducibility of both methods was excellent (with urine, antibody sandwich assay intraclass correlation coefficient = 0.9975 and with serum = 0.9949; correlation coefficient of the inhibition assay with urine = 0.9736 and with serum = 0.9850), that of the inhibition method was only fair to poor for the controls: urine = −0.0152, serum = 0.5595. Reproducibility was good for the controls using the sandwich method: urine = 0.7717, serum = 0.9470. Cross-reactivity was observed in specimens from patients infected with
Blastomyces dermatitidis
,
Paracoccidioides brasiliensis
, and
Penicillium marneffei
. In conclusion, the decreased specificity and inferior reproducibility with control specimens suggest that the inhibition assay has poorer precision toward the lower end of the detection range.
Sequence Analysis of the
ank
Gene of Granulocytic Ehrlichiae
Authors: Massung Robert F. , Owens Jessica H. , Ross David , Reed Kurt D. , Petrovec Miroslav , Bjoersdorff Anneli , Coughlin Richard T. , Beltz Gerald A. , Murphy Cheryl I. 
Published: December 2019

ABSTRACT

The
ank
gene of the agent of human granulocytic ehrlichiosis (HGE) codes for a protein with a predicted molecular size of 131.2 kDa that is recognized by serum from both dogs and humans infected with granulocytic ehrlichiae. As part of an effort to assess the phylogenetic relatedness of granulocytic ehrlichiae from different geographic regions and in different host species, the
ank
gene was PCR amplified and sequenced from a variety of sources. These included 10 blood specimens from patients with confirmed human granulocytic ehrlichiosis (three from New York, four from Wisconsin, two from Slovenia, and one from Sweden). Also examined was a canine granulocytic ehrlichia sample obtained from Minnesota,
Ehrlichia equi
from California,
Ehrlichia phagocytophila
from Sweden, and the granulocytic ehrlichia isolate USG3. The sequences showed a high level of homology (>95.5% identity), with the lowest homology occurring between a New York HGE agent and the Swedish
E. phagocytophila
. Several 3-bp deletions and a variable number of 51- and 81-bp direct repeats were noted. Although the North American HGE sequences showed the highest conservation (>98.1% identity), phylogenetic analyses indicated that these samples represent two separate clades, one including the three New York HGE samples and the USG3 strain and another with the Wisconsin HGE and Minnesota canine sequences. Two of the New York samples and the USG3 strain showed 100% identity over the entire 3,696-bp product. Likewise, three of the Wisconsin human samples and the Minnesota dog sample were identical (3,693 bp). Whereas phylogenetic analysis showed that the
E. equi
sequence was most closely related to the Upper Midwest samples, analysis of the repeat structures showed it to be more similar to the European samples. Overall, the genetic analysis based on the
ank
gene showed that the granulocytic ehrlichiae are closely related, appear to infect multiple species, and can be grouped into at least three different clades, two North American and one European.
Mapping of IS
6110
Insertion Sites in Two Epidemic Strains of
Mycobacterium tuberculosis
Authors: Beggs Marjorie L. , Eisenach Kathleen D. , Cave M. Donald 
Published: December 2019

ABSTRACT

A widely distributed strain designated 210 was identified in a study of the diversity of
Mycobacterium tuberculosis
DNA fingerprints from three geographically separate states in the United States. This strain is characterized by a 21-band fingerprint pattern when probed with IS
6110
, and the pattern is similar to that displayed by strains designated W. Intracellular growth of strain 210 isolates in human macrophages is significantly faster than that of isolates from other clusters or nonclustered isolates. The purpose of this study was to identify the sites of IS
6110
insertions in strain 210 and compare these to IS
6110
insertion sites in strain W. Our hypothesis is that an IS
6110
insertion site(s) could possibly be responsible for a strain's increased capacity for transmission and/or replication. In this report, the insertion sites in strains 210 and W are described and referenced to their location in the
M. tuberculosis
H37Rv genome sequence. The W and 210 strains have 17 identical sites of IS
6110
insertion and additional sequence not found in H37Rv but present in other clinical isolates. The IS
6110
insertion site in the 36-bp direct repeat (DR) region of strains 210 and W has 15 spacers in the left flanking region. The DR region on the right side of IS
6110
has been deleted. Five sites of insertion in strain 210 not found in strain W are described, as well as two unique sites in strain W. One copy of IS
6110
was found to reside 55 bp in the
ctpD
gene. This gene is expressed, indicating that IS
6110
can provide a promoter sequence for the transcription of genes.
Molecular Analysis of H Antigens Reveals that Human Diarrheagenic
Escherichia coli
O26 Strains That Carry the
eae
Gene Belong to the H11 Clonal Complex
Authors: Zhang Wen-Lan , Bielaszewska Martina , Bockemühl Jochen , Schmidt Herbert , Scheutz Flemming , Karch Helge 
Published: December 2019

ABSTRACT

Fifty-seven
Escherichia coli
O26 strains isolated from patients in six countries were investigated by PCR restriction fragment length polymorphism (RFLP) analysis of the flagellin-encoding (
fliC
) gene (
fliC
RFLP analysis). The strains were determined by serotyping to belong to five different H types or were nonmotile. The
fliC
RFLP analysis revealed only two different patterns among the 57 strains. One
fliC
RFLP pattern was displayed by 54 strains and was identical to that of
E. coli
H11 reference strain Su4321-41. The other
fliC
RFLP pattern was observed for three strains and was identical to that for
E. coli
H32 reference strain K10. The 54 strains with the H11
fliC
RFLP pattern included 22 strains of serotype O26:H11, 23 nonmotile strains, and 9 strains that were initially serotyped as H2, H8, H21, and H32 but that were confirmed to express H11 by repeat serotyping. All 54 strains with the H11
fliC
RFLP pattern contained the attaching-and-effacing (
eae
) gene. The three strains with the H32
fliC
RFLP pattern belonged to serotype O26:H32, and all were
eae
negative. The
fliC
genes of 14 selected
E. coli
O26:H11 strains isolated between 1964 and 1999 had identical nucleotide sequences. Our results demonstrate that
E. coli
O26 strains that carry the
eae
gene belong exclusively to the H11 clonal complex. Since there were no H11
fliC
allelic variations among the O26 strains tested,
E. coli
O26:H11 may have emerged recently. The
fliC
PCR-RFLP test is a reliable, easy-to-perform, and rapid method for determination of the H types of
E. coli
O26 isolates.
Clinical Significance and Antibiotic Resistance Patterns of
Leminorella
spp., an Emerging Nosocomial Pathogen
Authors: Blekher Lidia , Siegman-Igra Yardena , Schwartz David , Berger Stephen A. , Carmeli Yehuda 
Published: December 2019

ABSTRACT

Although
Leminorella
spp., members of the family
Enterobacteriaceae
, were previously isolated from feces and urine specimens, clinical correlates have not been studied. We conducted a retrospective study to investigate the clinical significance and disease spectrum of these organisms, as well as their antibiotic susceptibility patterns. Identification and susceptibility testing were performed by an automated system. Eighteen cases were identified retrospectively during a 28-month period (1/97 to 4/99), representing an incidence of 11 cases per 100,000 patient admissions. The medical records of 14 patients were reviewed. The average patient age was 67 years, and 78% were males. Patients had multiple and diverse underlying conditions which might have predisposed them to infection.
Leminorella
spp. were classified as definite pathogens in 43% of the cases, probable pathogens in 29%, and possible pathogens in 21%. In one case of asymptomatic bacteriuria, the isolate had no clinical significance. All infections but one were nosocomial. Clinical syndromes included urinary tract infection in six patients, surgical site infection in three patients, and primary bacteremia, peritonitis, respiratory tract infection, and soft tissue infection in one patient each. Isolates were uniformly susceptible to imipenem. Other beta-lactam agents had poor activity against the isolates. We conclude that
Leminorella
spp. are significant nosocomial pathogens that are capable of causing a variety of clinical syndromes and are resistant to multiple antibiotic agents.
Urine
d
-Arabinitol/
l
-Arabinitol Ratio in Diagnosis of Invasive Candidiasis in Newborn Infants
Authors: Sigmundsdóttir Gudrún , Christensson Bertil , Björklund Lars J. , Håkansson Kristina , Pehrson Christina , Larsson Lennart 
Published: December 2019

ABSTRACT

Infants treated in neonatal intensive care units suffer an increased risk for invasive candidiasis, but the diagnosis is sometimes difficult.
d
-arabinitol is a metabolite of most pathogenic
Candida
species. An elevated urine
d
-arabinitol/
l
-arabinitol (DA/LA) ratio is a sensitive sign of invasive candidiasis in children with cancer, but the method has not been previously evaluated for newborn infants. We therefore enrolled 117 infants in a neonatal intensive care unit, and 411 urine samples were obtained on filter paper. The DA/LA ratio was measured by gas chromatography-mass spectrometry. For 81 infants with no suspicion of superficial or invasive candidiasis, the urine DA/LA ratio was 2.7 ± 0.7 (mean ± standard deviation [SD]). The upper cutoff level was set at 4.8 (mean plus 3 SD). Of 22 infants with mucocutaneous candidiasis and not given systemic antifungal treatment, two had elevated DA/LA ratios, which normalized after removal of intravascular catheters. Eight other infants were given empiric antifungal treatment but had negative cultures; five of these had repeatedly elevated DA/LA ratios. Six infants with culture-positive invasive candidiasis all had one or more samples with elevated ratios. For seven infants, three with suspected and four with confirmed invasive candidiasis (for which follow-up samples were available), ratios normalized during antifungal treatment. In conclusion, urine DA/LA ratio determination is a rapid test and can be used for newborns. It is possibly more sensitive than fungal blood cultures in the diagnosis of invasive candidiasis and can also be used for monitoring the effect of antifungal treatment.
Molecular Identification of
Dialister pneumosintes
in Subgingival Plaque of Humans
Authors: Doan N. , Contreras A. , Flynn J. , Slots J. , Chen C. 
Published: December 2019

ABSTRACT

Dialister pneumosintes
is a nonfermentative, anaerobic, gram-negative rod that grows with small, circular, transparent, shiny, smooth colonies on blood agar. Even though
D. pneumosintes
has been recovered from deep periodontal pockets, little is known about the relationship between the organism and destructive periodontal disease. This study describes a rapid PCR method to identify
D. pneumosintes
in periodontal samples. The PCR identification method detected as little as 10 pg of
D. pneumosintes
DNA or about 1 to 10 cells without nonspecific amplification of various periodontopathic bacteria. Twelve of 22 subgingival samples from adult periodontitis lesions yielded
D. pneumosintes
either by culture or by PCR identification. In culture-positive samples,
D. pneumosintes
averaged 3.9% (0.001 to 10.8%) of total isolates. Studies are needed to delineate virulence factors of
D. pneumosintes
pertinent to periodontal disease.
Diagnostic Utility of a Multiplex Herpesvirus PCR Assay Performed with Cerebrospinal Fluid from Human Immunodeficiency Virus-Infected Patients with Neurological Disorders
Authors: Quereda C. , Corral I. , Laguna F. , Valencia M. E. , Tenorio A. , Echeverria J. E. , Navas E. , Martín-Dávila P. , Moreno A. , Moreno V. , Gonzalez-Lahoz J. M. , Arribas J. R. , Guerrero A. 
Published: December 2019

ABSTRACT
We used a multiplex nested-PCR assay for the simultaneous detection in cerebrospinal fluid (CSF) of five human herpesviruses (HVs) (cytomegalovirus [CMV], Epstein-Barr virus [EBV], varicella-zoster virus [VZV], herpes simplex virus [HSV], and human herpesvirus 6 [HHV-6]) in a clinical evaluation of human immunodeficiency virus (HIV)-infected patients with neurological disorders. This method, which has the advantages of being rapid and economical, would be of particular interest for the diagnosis of neurological syndromes caused by more than one HV. We studied 251 CSF samples from 219 patients. HV DNA was demonstrated in 93 (37%) of the CSF samples (34% of the patients). CMV was the HV most frequently detected in our patients (25%), while EBV, VZV, HSV, and HHV-6 DNAs were present in significantly fewer cases (7, 4, 3, and 1%, respectively). When results were compared with the final etiological diagnoses of the patients, the multiplex HV PCR showed high specificity for the diagnosis of CMV and VZV neurological diseases and for cerebral lymphoma (0.95, 0.97, and 0.99, respectively). The sensitivity of the assay was high for CMV disease (0.87), was low for cerebral lymphoma (0.33), and was not evaluable for VZV disease due to the small number of patients with this diagnosis. Nevertheless, detection of VZV DNA had possible diagnostic value in four of the nine cases, and EBV DNA amplification always predicted the diagnosis of cerebral lymphoma in patients with cerebral masses. Detection of HSV DNA was frequently associated with CMV amplification and fatal encephalitis. HHV-6 was not considered to have a pathogenetic role in the three cases in which it was detected. This multiplex HV PCR assay is a specific and clinically useful method for the evaluation of HIV-infected patients with neurological disorders related to HV.
Rapid Detection of
Campylobacter jejuni
in Stool Specimens by an Enzyme Immunoassay and Surveillance for
Campylobacter upsaliensis
in the Greater Salt Lake City Area
Authors: Hindiyeh Musa , Jense Sandra , Hohmann Sheri , Benett Hilary , Edwards Cheryl , Aldeen William , Croft Ann , Daly Judy , Mottice Susan , Carroll Karen C. 
Published: December 2019

ABSTRACT

The Alexon-Trend, Inc. (Ramsey, Minn.), ProSpecT
Campylobacter
microplate assay was compared with culture on a Campy-CVA plate (Remel, Lenexa, Kans.) and blood-free campylobacter agar with cefoperazone (20 μg/ml), amphotericin B (10 μg/ml), and teicoplanin (4 μg/ml) (CAT medium; Oxoid Limited, Hampshire, England) with 631 patient stool samples. The CAT medium was used to isolate
Campylobacter upsaliensis
. The enzyme immunoassay (EIA) had a sensitivity and a specificity of 89 and 99%, respectively, and the positive and negative predictive values were 80 and 99%, respectively. Even though we extensively looked for
C. upsaliensis
in stool samples from patients from the greater Salt Lake City area, we did not isolate this species during the study period. The overall excellent specificity of the EIA allows rapid detection and treatment of positive patients; however, a negative result should be confirmed by culture when clinical suspicion is high.
Isolation of
Legionella oakridgensis
from Two Patients with Pleural Effusion Living in the Same Geographical Area
Authors: Lo Presti François , Riffard Serge , Jarraud Sophie , Le Gallou Florence , Richet Hervé , Vandenesch François , Etienne Jerome 
Published: December 2019

ABSTRACT

Two cases of Legionnaire's disease caused by
Legionella oakridgensis
were diagnosed at the university hospital in Nantes, France. The two patients' isolates were identified by means of phenotyping and genotyping methods. Epidemiological investigations concluded that the first case was hospital acquired while the second case was considered community acquired.
Repression of the A8L Gene, Encoding the Early Transcription Factor 82-Kilodalton Subunit, Inhibits Morphogenesis of Vaccinia Virions
Authors: Hu Xiaolei , Wolffe Elizabeth J. , Weisberg Andrea S. , Carroll Lawrence J. , Moss Bernard 
Published: December 2019

ABSTRACT

The vaccinia virus early transcription factor (VETF) is a DNA binding protein comprised of 70- and 82-kDa subunits encoded by the D6R and A8L genes, respectively. A previous investigation suggested a novel role for the 70-kDa subunit in the morphogenesis of vaccinia virus particles. The principal objectives of the present study were to determine if the 82-kDa subunit of VETF is also required for morphogenesis and, if so, whether the block occurs before or after the incorporation of the genome into the assembling virus particle. To address these and other questions, we constructed and characterized a conditionally lethal recombinant vaccinia virus in which the A8L gene is stringently repressed by the
Escherichia coli lac
operator system. The amount of 82-kDa protein synthesized could be regulated by the amount of inducer: from undetectable to higher than normal levels. Virus replication, as determined by plaque formation or virus yield upon synchronous infection, was dependent on inducer. Nevertheless, de novo synthesis of the 82-kDa subunit was not required for viral early, intermediate, and late gene expression or DNA replication. Overexpression of the A8L gene alone, produced by high concentrations of inducer, inhibited viral late protein synthesis, whereas overexpression of the D6R gene alone or both VETF genes simultaneously had little inhibitory effect. Laser confocal fluorescence and quantitative electron microscopic analyses revealed that immature and DNA-containing intermediate stage particles accumulated in the absence of inducer, indicating that the A8L protein has a role in morphogenesis of the core and subsequent events.
Transgenic Mice Expressing Human Immunodeficiency Virus Type 1 in Immune Cells Develop a Severe AIDS-Like Disease
Authors: Hanna Zaher , Kay Denis G. , Cool Marc , Jothy Serge , Rebai Najet , Jolicoeur Paul 
Published: December 2019

ABSTRACT
We have constructed transgenic (Tg) mice expressing the entire human immunodeficiency virus type 1 (HIV-1) coding sequences in cells targeted by HIV-1 infection in humans. These Tg mice developed a severe AIDS-like disease leading to early death (<1 month). They developed muscle wasting, severe atrophy and fibrosis of lymphoid organs, tubulointerstitial nephritis, and lymphoid interstitial pneumonitis. In addition the expression of RANTES was increased in various tissues of these Tg mice relative to that in the normal controls. Disease appearance was correlated with the levels of transgene expression. The numerous pathologies observed in these mice are remarkably similar to those observed in human AIDS and, more specifically, in pediatric AIDS.
Hepatitis B Virus X Protein Interferes with Cellular DNA Repair
Authors: Becker Sherry A. , Lee Teh-Hsiu , Butel Janet S. , Slagle Betty L. 
Published: December 2019

ABSTRACT
The hepatitis B virus X protein (HBx) is a broadly acting transactivator implicated in the development of liver cancer. Recently, HBx has been reported to interact with several different cellular proteins, including our report of its binding to XAP-1, the human homolog of the simian repair protein UVDDB. In the present study, several HBx mutants were used to localize the minimal domain of HBx required for binding to XAP-1/UVDDB to amino acids 55 to 101. The normal function of XAP-1/UVDDB is thought to involve binding to damaged DNA, the first step in nucleotide excision repair (NER); therefore, we hypothesized that this interaction may affect the cell’s capacity to correct lesions in the genome. When tested in two independent assays that measure NER (unscheduled DNA synthesis and host cell reactivation), the expression of HBx significantly inhibited the ability of cells to repair damaged DNA. Under the assay conditions, HBx was expressed at a level similar to that previously observed during natural viral infection and was able to transactivate several target reporter genes. These results are consistent with a model in which HBx acts as a cofactor in hepatocarcinogenesis by preventing the cell from efficiently repairing damaged DNA, thus leading to an accumulation of DNA mutations and, eventually, cancer. An adverse effect on cellular DNA repair processes suggests a new mechanism by which a tumor-associated virus might contribute to carcinogenesis.