bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Etest Method for Determining Caspofungin (MK-0991) Susceptibilities of 726 Clinical Isolates of
Candida
Species
Authors: Pfaller M. A. , Messer S. A. , Mills K. , Bolmström A. , Jones R. N. 
Published: July 2002

ABSTRACT

The performance of the Etest for testing the susceptibilities to caspofungin (MK-0991) of 726 isolates of
Candida
spp. was assessed against the National Committee for Clinical Laboratory Standards (NCCLS) microdilution broth method. The NCCLS method employed RPMI 1640 broth medium, and MICs were read after incubation for 48 h at 35°C. MICs were determined by Etest for all 726 isolates with RPMI agar containing 2% glucose (RPG) and were read after incubation for 48 h at 35°C. The
Candida
isolates included
Candida albicans
(
n
= 486),
Candida glabrata
(
n
= 96),
Candida tropicalis
(
n
= 51),
Candida parapsilosis
(
n
= 47),
Candida krusei
(
n
= 11),
Candida lusitaniae
(
n
= 2), and
Candida guilliermondii
(
n
= 33). In addition, a subset of 314 isolates were also tested by Etest using Casitone agar (CAS) and antibiotic medium 3 agar (AM3). The Etest results obtained using RPG correlated well with reference MICs. Overall agreement was 94% with RPG, 82% with CAS, and 79% with AM3. When RPG was used, agreement ranged from 79% for
C. parapsilosis
to 100% for
C. krusei
,
C. lusitaniae
, and
C. guilliermondii
. When CAS was used, agreement ranged from 0% for
C. lusitaniae
to 100% for
C. glabrata
. With AM3, agreement ranged from 0% for
C. lusitaniae
to 100% for
C. guilliermondii
. All three media supported growth of each of the
Candida
species. Etest results were easy to read, with sharp zones of inhibition. In most instances (75%) where a discrepancy was observed between the Etest and the reference method, the Etest MIC was lower. The Etest method using RPG appears to be useful for determining caspofungin susceptibilities of
Candida
species.
Isolations of
Leclercia adecarboxylata
from a Patient with a Chronically Inflamed Gallbladder and from a Patient with Sepsis without Focus
Authors: de Baere Thierry , Wauters Georges , Huylenbroeck Anne , Claeys Geert , Peleman Renaat , Verschraegen Gerda , Allemeersch Daniel , Vaneechoutte Mario 
Published: July 2002

ABSTRACT

Leclercia adecarboxylata
was isolated from a patient with a chronically inflamed gallbladder, together with
Enterococcus
sp. The organism was considered clinically significant and was susceptible to all antibiotics tested. Another strain of
L. adecarboxylata
was cultured from blood, together with
Escherichia hermannii
and
E. faecalis
, from a patient with sepsis.
Identification of
Actinobacillus actinomycetemcomitans
Serotypes by Multiplex PCR
Authors: Suzuki Nao , Nakano Yoshio , Yoshida Yasuo , Ikeda Daisuke , Koga Toshihiko 
Published: July 2002

ABSTRACT

Oligonucleotide primers specific for gene clusters involved in the biosynthesis of serotype-specific polysaccharide antigens were designed to identify
Actinobacillus actinomycetemcomitans
serotypes a to e using the multiplex PCR. This method may be useful for serotype-specific genotyping rapidly and directly from clinical samples containing various organisms.
Genomic Diversity of Group A Rotavirus Strains Infecting Humans in Eastern India
Authors: Das Soma , Sen A. , Uma G. , Varghese V. , Chaudhuri S. , Bhattacharya S. K. , Krishnan T. , Dutta P. , Dutta D. , Bhattacharya M. K. , Mitra U. , Kobayashi N. , Naik T. N. 
Published: July 2002

ABSTRACT
Between 1998 and 2000, a total of 266 samples were found positive for group A rotaviruses by RNA electrophoresis. Samples were collected from patients admitted to two leading hospitals in Calcutta. Serotyping could be done only with 22% of the positive samples, leaving 78% untypeable. The G (VP7 genotypes) and P (VP4 genotypes) types were determined for 159 samples by reverse transcription and multiplex PCR. The predominant genotype was G1P[8] (20%), followed by G2P[4] (15%) and G4P[8] (6%). A number of uncommon genotypes, G1P[4] (4%), G2P[8] (2.5%), G2P[6] (0.6%), G4P[4] (2.5%), and G4P[6] (1.25%), were also detected during this study period. Twenty two percent of specimens showed mixed infections, 38 (24%) of the total samples remained untypeable for either VP7 or VP4, while only 4 (2.5%) of the samples were untypeable for both genes. Eleven specimens collected from Manipur were also genotyped and revealed a very high degree of genomic reassortment.
Testing of Susceptibility of
Mycobacterium tuberculosis
to Pyrazinamide with the Nonradiometric BACTEC MGIT 960 System
Authors: Pfyffer Gaby E. , Palicova Frantiska , Rüsch-Gerdes Sabine 
Published: July 2002

ABSTRACT

The reliability of the novel BACTEC MGIT 960 pyrazinamide (PZA) kit (Becton Dickinson Microbiology Systems, Sparks, Md.) was assessed for testing of susceptibility of
Mycobacterium tuberculosis
to PZA. Results generated by the BACTEC MGIT 960 system (Becton Dickinson) were compared with those obtained with the BACTEC 460TB system. Extensive proficiency testing (phase I) and reproducibility testing (phase II) as well as susceptibility testing of blinded strains of
M. tuberculosis
from the Centers for Disease Control and Prevention (phase III) were performed prior to testing 58 strains isolated from clinical specimens (phase IV). After resolution of discrepant results obtained by the two BACTEC methods by two other laboratories which acted as independent arbiters (phase V), overall agreement of the BACTEC MGIT 960 system with the BACTEC 460TB system for PZA testing of phase IV strains was 96.6%. Between the two systems there was no statistically significant difference in time until results were obtained, i.e., 6.8 days (BACTEC MGIT 960) versus 5.4 days (BACTEC 460TB), the latter not counting the time required for a subculture with a growth index of 200, however. The new BACTEC MGIT PZA susceptibility testing procedure works equally well for inocula prepared from liquid (MGIT) and solid (Löwenstein-Jensen) cultures. PZA MGIT medium in plastic tubes yielded results equivalent to medium dispensed in glass tubes.
Detection, Subgroup Specificity, and Genotype Diversity of Rotavirus Strains in Children with Acute Diarrhea in Paraguay
Authors: Coluchi Norma , Munford Veridiana , Manzur Julio , Vazquez Cynthia , Escobar Mabel , Weber Ernesto , Mármol Perla , Rácz Maria Lucia 
Published: July 2002

ABSTRACT
Of a total of 220 stool specimens from children with acute diarrhea, mostly under the age of 3 years, collected in Paraguay between January 1999 and March 2000, 70 (31.8%) were found positive for rotaviruses (RV). Positive samples were characterized by electropherotyping and subgrouping. Sixty-one (87.1%) were classified as group A, subgroup II; one (1.4%) was classified as group A, subgroup I; six (8.6%) were group A, non-I non-II; and two (2.9%) were not tested. RV strains were G and P genotyped by reverse transcription-PCR. The following G types were detected: G4 (34.3%), G1 (21.4%), G2 (1.4%), and G9 (5.7%). Mixtures of human and animal genotypes were detected in 15 (21.4%) samples, and 11 samples (15.7%) were nontypeable. The following P types were detected: P[8] (48.6%), P[4] (1.4%), and P[1] (1.4%). A mixed type was found in 10% of samples, and an unexpectedly high percentage (38.6%) of nontypeable samples was found. The common human G- and P-type combinations P[8], G4 (15.7%) and P[8], G1 (14.2%) were detected. Mixed human and animal genotypes were observed as the following combinations: G4 + G5, G4 + G5 + G10, and G1 + G10 for G types and P[8]-P[1] for P types. The emerging G9 genotype was detected in four samples. These results show for the first time the diversity of RV circulating among children in Paraguay and contribute to the knowledge of this pathogen required to devise strategies to prevent diarrheal illness in this country. The finding of mixed genotypes may indicate interspecies transmission of RV between humans and animals.
Evaluation of the DipStreak, a New Device with an Original Streaking Mechanism for Detection, Counting, and Presumptive Identification of Urinary Tract Pathogens
Authors: Scarparo Claudio , Piccoli Paola , Ricordi Paolo , Scagnelli Mariuccia 
Published: July 2002

ABSTRACT

DipStreak is a new urine culture device with two types of agar attached back-to-back on a plastic paddle. It combines dip-slide technology and an original streaking inoculation mechanism, allowing for bacterial counting and colony isolation. The performance of the DipStreak device with two different medium formulations, CHROMagar and MacConkey media in study A and UriSelect 3 and MacConkey media in study B, was evaluated and compared to that of the reference streak method by using plates of cystine-lactose-electrolyte-deficient (CLED) agar, tryptic soy agar with 5% sheep blood, and UriSelect 3 medium. In study A, 2,000 urine specimens were processed and 511 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave the same detection rate, 99.7%. For the direct identification of
Escherichia coli
,
Proteus mirabilis
, and
Enterococcus
sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 97 and 93.4%, respectively. In study B, 3,000 urine specimens were processed and 714 cultures were found positive. The DipStreak device and the UriSelect 3 and CLED medium plates gave detection rates of 99.4, 99.9, and 99.2%, respectively. For the direct identification of
E. coli
,
P. mirabilis
, and
Enterococcus
sp. isolates, the DipStreak device and the UriSelect 3 medium plate showed overall sensitivities of 88 and 94.4%, respectively. In conclusion, the DipStreak device with both medium formulations represents an attractive and excellent screening method for the reliable detection, counting, and presumptive identification of urinary tract pathogens. It enables bedside urine inoculation and provides a valid means of transporting the sample back to the laboratory, decreasing drastically the rate of false-positive results due to bacterial overgrowth and reducing associated costs.
Expression of the Bundle-Forming Pilus by Enteropathogenic
Escherichia coli
Strains of Heterologous Serotypes
Authors: Gismero-Ordoñez Juana , Dall'Agnol Monique , Trabulsi Luiz R. , Girón Jorge A. 
Published: July 2002

ABSTRACT

The production of the bundle-forming pilus subunit BfpA was investigated in 44 enteropathogenic
Escherichia coli
(EPEC) diarrheal isolates after growth on several conventional bacteriological media. In all, the use of brucella agar supplemented with 4.5 mM sodium bisulfite resulted in a higher detection of BfpA-producing EPEC.
Pneumonia Due toBordetella bronchisepticain a Cystic Fibrosis Patient: 16S rRNA Sequencing for Diagnosis Confirmation
Authors: Wallet Frédéric , Perez Thierry , Armand Sylvie , Wallaert Benoit , Courcol René J. 
Published: July 2002

ABSTRACTBordetella bronchisepticawas identified as an unusual etiologic agent of pulmonary recurrent exacerbations and pneumonia in a cystic fibrosis (CF) patient by utilizing a 16S rRNA molecular kit in our hospital's clinical laboratory. This method appears to be a useful approach for identifying new emerging CF pathogens when discrepancies exist between phenotypical tests.
Functional Analysis of theenvOpen Reading Frame in Human Endogenous Retrovirus IDDMK1,222 Encoding Superantigen Activity
Authors: Lapatschek Matthias , Dürr Susanne , Löwer Roswitha , Magin Christine , Wagner Hermann , Miethke Thomas 
Published: July 2002

ABSTRACTMice harbor a family of endogenous retroviruses, the mouse mammary tumor viruses (MMTV), which encode superantigens. These superantigens are responsible for the deletion of T cells expressing certain Vβ chains of the T-cell receptor in the thymus. Human T cells are able to recognize MMTV-encoded superantigens presented by human major histocompatibility complex class II-positive cells. Owing to this and to the similarity of the human and murine immune systems, it was speculated that human endogenous retroviruses might also code for superantigens. Recently, it was reported that a proviral clone (IDDMK1,222) of the human endogenous retrovirus family HTDV/HERV-K encodes a superantigen. The putative superantigen gene was located within theenvregion of the virus. Stimulated by these findings, we amplified by PCR and cloned into eucaryotic expression vectors open reading frames (ORFs) which were identical or very similar to IDDMK1,222. When we transfected these vectors into A20 cells, a murine B-cell lymphoma, we were able to demonstrate mRNA expression and protein production. However, we did not find any evidence that the ORF stimulated human or murine T cells in a Vβ-specific fashion, the most prominent feature of superantigens.
The Murine Gammaherpesvirus 68 v-Cyclin Is a Critical Regulator of Reactivation from Latency
Authors: van Dyk Linda F. , Virgin Herbert W. , Speck Samuel H. 
Published: July 2002

ABSTRACTGamma-2 herpesviruses encode a homolog of mammalian D-type cyclins. The v-cyclin encoded by murine gammaherpesvirus 68 (γHV68) induces cell cycle progression and is an oncogene (L. F. van Dyk, J. L. Hess, J. D. Katz, M. Jacoby, S. H. Speck, and H. W. Virgin IV, J. Virol. 73:5110–5122, 1999). However, the role of the pro-proliferative v-cyclins in gamma-2 herpesvirus pathogenesis is not known. Here we report the generation and characterization of a γHV68 v-cyclin mutant (v-cyclin.LacZ) that is unable to express a functional v-cyclin protein. Notably, although the γHV68 v-cyclin is expressed from an early-late lytic transcript, v-cyclin.LacZ replicated normally in fibroblasts in vitro and during acute infection in the spleen, liver, and lungs in vivo. Moreover, v-cyclin.LacZ exhibited wild-type (wt) virulence in mice with severe combined immunodeficiency. In addition, in a model of γHV68-induced chronic disease in mice lacking the gamma interferon receptor (IFNγR−/−), v-cyclin.LacZ virus was similar to wt γHV68 in terms of the incidence of mortality and vasculitis. Further analysis revealed that the frequencies of splenocytes and peritoneal cells harboring the latent γHV68 genome in normal and B-cell-deficient mice infected with wt γHV68 or v-cyclin.LacZ were very similar. However, v-cyclin.LacZ was significantly compromised in its capacity to reactivate from latency. This phenotype was conclusively mapped to the v-cyclin gene by (i) generating a marker rescue virus (v-cyclin.MR) from the v-cyclin.LacZ mutant, which restored the frequency of cells in which virus reactivated from latency to the levels observed with wt γHV68; and (ii) generating a second v-cyclin mutant virus containing a translation stop codon within the v-cyclin gene (v-cyclin.stop), which was compromised in reactivation from latency. These studies demonstrate that despite expression as a lytic cycle gene, the pro-proliferative γHV68 v-cyclin is not required for γHV68 replication either in vitro or during acute infection in vivo but rather is a critical determinant of reactivation from latency.
Identification of Mouse Hepatitis Virus Papain-Like Proteinase 2 Activity
Authors: Kanjanahaluethai Amornrat , Baker Susan C. 
Published: July 2002

ABSTRACT

Mouse hepatitis virus (MHV) is a 31-kb positive-strand RNA virus that is replicated in the cytoplasm of infected cells by a viral RNA-dependent RNA polymerase, termed the replicase. The replicase is encoded in the 5′-most 22 kb of the genomic RNA, which is translated to produce a polyprotein of >800 kDa. The replicase polyprotein is extensively processed by viral and perhaps cellular proteinases to give rise to a functional replicase complex. To date, two of the MHV replicase-encoded proteinases, papain-like proteinase 1 (PLP1) and the poliovirus 3C-like proteinase (3CLpro), have been shown to process the replicase polyprotein. In this report, we describe the cloning, expression, and activity of the third MHV proteinase domain, PLP2. We show that PLP2 cleaves a substrate encoding the first predicted membrane-spanning domain (MP1) of the replicase polyprotein. Cleavage of MP1 and release of a 150-kDa intermediate, p150, are likely to be important for embedding the replicase complex in cellular membranes. Using an antiserum (anti-D11) directed against the C terminus of the MP1 domain, we verified that p150 encompasses the MP1 domain and identified a 44-kDa protein (p44) as a processed product of p150. Pulse-chase experiments showed that p150 is rapidly generated in MHV-infected cells and that p44 is processed from the p150 precursor. Protease inhibitor studies revealed that unlike 3CLpro activity, PLP2 activity is not sensitive to cysteine protease inhibitor E64d. Furthermore, coexpression studies using the PLP2 domain and a substrate encoding the MP1 cleavage site showed that PLP2 acts efficiently in
trans
. Site-directed mutagenesis studies confirmed the identification of cysteine 1715 as a catalytic residue of PLP2. This study is the first to report enzymatic activity of the PLP2 domain and to demonstrate that three distinct viral proteinase activities process the MHV replicase polyprotein.
Human Immunodeficiency Virus Type 1- and Cytomegalovirus-Specific Cytotoxic T Lymphocytes Can Persist at High Frequency for Prolonged Periods in the Absence of Circulating Peripheral CD4+T Cells
Authors: Spiegel Hans M. L. , Ogg Graham S. , DeFalcon Elizabeth , Sheehy Megan E. , Monard Simon , Haslett Patrick A. J. , Gillespie Geraldine , Donahoe Sean M. , Pollack Henry , Borkowsky William , McMichael Andrew J. , Nixon Douglas F. 
Published: July 2002

ABSTRACTCD4+T cells are thought to be critical in the maintenance of virus-specific CD8+cytotoxic T-cell (CTL) responses. In human immunodeficiency virus type 1 (HIV-1) infection, a selective decline in HIV-1-specific CTL as the CD4+T-cell count decreases has been reported. Using HLA-peptide tetrameric complexes, we show the presence at high frequency of HIV-1- and cytomegalovirus-specific CD8+T cells when the peripheral CD4+T-cell count was low or zero in three HIV-1-infected patients. No direct virus-specific CD8+-mediated effector activity was seen in these subjects, suggesting antigen unresponsiveness, although tetramer-sorted cells could be expanded in vitro in the presence of interleukin-2 into responsive effector cells. Thus, virus-specific CD8+T cells can be maintained in the peripheral circulation at high frequency in the absence of circulating peripheral CD4+T cells, but these cells may lack direct effector activity. Strategies designed to overcome this antigen unresponsiveness may be of value in therapies for the treatment of AIDS.
Recombinant Measles Viruses Efficiently Entering Cells through Targeted Receptors
Authors: Schneider Urs , Bullough Frances , Vongpunsawad Sompong , Russell Stephen J. , Cattaneo Roberto 
Published: July 2002

ABSTRACT
We sought proof of principle that one of the safest human vaccines, measles virus Edmonston B (MV-Edm), can be genetically modified to allow entry via cell surface molecules other than its receptor CD46. Hybrid proteins consisting of the epidermal growth factor (EGF) or the insulin-like growth factor 1 (IGF1) linked to the extracellular (carboxyl) terminus of the MV-Edm attachment protein hemagglutinin (H) were produced. The standard H protein gene was replaced by one coding for H/EGF or H/IGF1 in cDNA copies of the MV genome. Recombinant viruses were rescued and replicated to titers approaching those of the parental strain. MV displaying EGF or IGF1 efficiently entered CD46-negative rodent cells expressing the human EGF or the IGF1 receptor, respectively, and the EGF virus caused extensive syncytium formation and cell death. Taking advantage of a factor Xa protease recognition site engineered in the hybrid H proteins, the displayed domain was cleaved off from virus particles, and specific entry in rodent cells was abrogated. These studies prove that MV can be engineered to selectively eliminate cells expressing a targeted receptor and provide insights into the mechanism of MV entry.
TT Virus Infection in Nonhuman Primates and Characterization of the Viral Genome: Identification of Simian TT Virus Isolates
Authors: Abe Kenji , Inami Tomoko , Ishikawa Koichi , Nakamura Shin , Goto Shunji 
Published: July 2002

ABSTRACT
Newly discovered TT virus (TTV) is widely distributed in human populations. To understand more about the relationship between TTV and its hosts, we tested 400 sera from various nonhuman primates for the presence of TTV DNA by PCR assay. We collected serum samples from 24 different species of nonhuman primates. TTV DNA was determined by PCR with primers designed from the 5′-end region of the TTV genome. Nucleotide sequencing and phylogenetic analysis of viral genomes were also performed. TTV DNA was detected in 87 of 98 (89%) chimpanzees and 3 of 21 (14%) crab-eating macaques. Nucleotide sequences of the PCR products obtained from both animals were 80 to 100% identical between two species. In contrast, the sequences differed from TTV isolates in humans by 24 to 33% at the nucleotide level and 36 to 50% at the amino acid level. Phylogenetic analysis demonstrated that all TTV isolates obtained from simians were distinct from the human TTV isolates. Furthermore, TTV in simians, but not in humans, was classified into three different genotypes. Our results indicate that TTV in simians represents a group different from, but closely related to, TTV in humans. From these results, we tentatively named this TTV simian TTV (s-TTV). The existence of the s-TTV will be important in determining the origin, nature, and transmission of human TTV and may provide useful animal models for studies of the infection and pathogenesis of this new DNA virus.
Infection of Primary Human Monocytes by Epstein-Barr Virus
Authors: Savard Martin , Bélanger Carole , Tardif Mélanie , Gourde Pierrette , Flamand Louis , Gosselin Jean 
Published: July 2002

ABSTRACTPrevious studies have reported that infection of monocytes by viruses such as cytomegalovirus and human immunodeficiency virus weakens host natural immunity. In the present study, we demonstrated the capability of Epstein-Barr virus (EBV) to infect and replicate in freshly isolated human monocytes. Using electron microscopy analysis, we observed the presence of EBV virions in the cytoplasm and nuclei of approximately 20% of monocytes. This was confirmed by Southern blot analysis of EBV genomic DNA sequences in isolated nuclei from monocytes. Infection of monocytes by EBV leads to the activation of the replicative cycle. This was supported by the detection of immediate-early lytic mRNA BZLF-1 transcripts, and by the presence of two early lytic transcripts (BALF-2, which appears to function in DNA replication, and BHRF-1, also associated with the replicative cycle). The late lytic BcLF-1 transcripts, which code for the major nucleocapsid protein, were also detected, as well as EBNA-1 transcripts. However, attempts to detect EBNA-2 transcripts have yielded negative results. Viral replication was also confirmed by the release of newly synthesized infectious viral particles in supernatants of EBV-infected monocytes. EBV-infected monocytes were found to have significantly reduced phagocytic activity, as evaluated by the quantification of ingested carboxylated fluoresceinated latex beads. Taken together, our results suggest that EBV infection of monocytes and alteration of their biological functions might represent a new mechanism to disrupt the immune response and promote viral propagation during the early stages of infection.