bluuebunny/crossref_metadata_2025_split huggingface dataset:
Annexin 1 localisation in tissue eosinophils as detected by electron microscopy
Authors: Oliani Sonia M. , Damazo Amilcar S. , Perretti Mauro 
Published: January 2003

Background: Human and rodent leukocytes express high levels of the glucocorticoid‐inducible protein annexin 1 (ANXA1) (previously referred to as lipocortin 1). Neutrophils and monocytes have abundant ANXA1 levels.Aim: We have investigated, for the first time, ANXA1 ultrastructural expression in rat eosinophils and compared it with that of extravasated neutrophils. The effect of inflammation (carrageenin peritonitis) was also monitored.Methods: Electron microscopy was used to define the sub‐cellular localisation of ANXA1 in rat eosinophils and neutrophils extravasated in the mesenteric tissue. A pair of antibodies raised against the ANXA1 N‐terminus (i.e. able to recognise intact ANXA1, termed LCPS1) or the whole protein (termed LCS3) was used to perform the ultrastructural analysis.Results: The majority of ANXA1 was localised in the eosinophil cytosol (~60%) and nucleus (30‐40%), whereas a small percentage was found on the plasma membrane (< 10%). Within the cytosol, the protein was equally distributed in the matrix and in the granules, including those containing the typical crystalloid. The two anti‐ANXA1 antibodies gave similar results, with the exception that LCPS1 gave a lower degree of immunoreactivity in the plasma membrane. Inflammation (i.e. carrageenin injection) produced a modest increase in eosinophil‐associated ANXA1 reactivity (significant only in the cytoplasm compartment). Extravasated neutrophils, used for comparative purposes, displayed a much higher degree of immunoreactivity for the protein.Conclusion: We describe for the first time ANXA1 distribution in rat eosinophil by ultrastructural analysis, and report a different protein mobilisation from extravasated neutrophils, at least in this acute model of peritonitis.
Effects of methotrexate upon inflammatory parameters induced by carrageenan in the mouse model of pleurisy
Authors: Dalmarco Eduardo Monguilhott , Fröde Tânia Silvia , Medeiros Yara Santos 
Published: January 2003

Background: The model of pleurisy induced by carrageenan exhibits a biphasic response (4 and 48 h) and permits the quantification of exudate, cell migration and certain enzymes such as myeloperoxidase (MPO) and adenosine‐deaminase (ADA) that are markers of activated leukocytes.Aims: The present study evaluates whether there exists, in the pleurisy model, a significant inhibition of ADA and MPO enzymes, leukocyte kinetics and other markers of inflammation [nitric oxide (NO) levels, exudation] caused by methotrexate treatment by the intraperitoneal (i.p.) route.Methods: The pleurisy was induced by carrageenan (1%) in mice, and the parameters were analyzed 4 and 48 h after.Results: After the induction of inflammation (4 h), methotrexate (20 mg/kg, i.p., 24 h before pleurisy induction) inhibited the leukocyte infiltration
(p < 0.05),
NO levels and MPO activity (p < 0.01), but not ADA activity and fluid leakage (p > 0.05). Regarding the second phase of pleurisy (48 h), methotrexate (40 mg/kg, i.p., 0.5 h before pleurisy induction) inhibited the leukocyte infiltration (p < 0.05), fluid leakage, NO levels (p < 0.01),
and ADA and MPO activity (p < 0.05).Conclusions: These findings support the evidence that the acute administration of methotrexate has an important systemic anti‐inflammatory activity in the studied inflammatory model. This effect was due to a significant inhibition on both neutrophil and mononuclear cells, being less marked in relation to exudation 48 h after. In relation to the enzymes studied and to NO levels, the findings support the evidence that methotrexate inhibits both enzymes (MPO and ADA) from leukocytes at the site of injury, thus reflecting the activation of both neutrophils and lymphocytes, respectively. Furthermore, the inhibiting effect on NO in both phases of pleurisy induced by carrageenan (4 and 48 h) indicates that methotrexate acts on constitutive and/or inducible NO synthases by means of different cells of the pleural cavity.
Effect of soluble interleukin‐6 receptor α and interleukin‐6 secreted by polymorphonuclear leukocytes on tumor necrosis factor‐α expression and its production by peripheral blood mononuclear cells
Authors: Jablonska E. 
Published: January 2003

Background: It has recently been shown that soluble interleukin‐6 receptor (sIL‐6R) alone or complexed with interleukin (IL)‐6, besides their regulatory role in a wide variety of both normal and abnormal biologic reactions mediated by IL‐6, could be an effective stimulator of the cell function.Aims: The key question of the present study is whether the sIL‐6Rα or sIL‐6R with IL‐6 released by polymorphonuclear leukocytes (PMN) can influence cytokine secretion such as tumor necrosis factor‐α (TNF‐α) by peripheral blood mononuclear cells (PBMC), which together with PMN develop the inflammatory and immune response of a host.Methods: Cells were isolated from heparinized whole blood of healthy persons. The PMN were cultured for 1 h at
37°C in 5% CO2. After incubation, the culture supernatant of PMN was removed and was added to PBMC. The PBMC were cultured for 1 h at 37°C in the same conditions. In the culture supernatants and lysates of PMN, we examined the concentrations of sIL‐6R by enzyme‐linked immunosorbent assay (ELISA). TNF‐α was measured at both protein and mRNA levels. Protein levels were determined by ELISA. To examine TNF‐α mRNA expression, we isolated mRNA from PBMC after culture, using TRIZOL Reagent. The quantity of mRNA TNF‐α was determined by the Quantikine mRNA assay.Results and conclusion: The results obtained revealed that sIL‐6R with IL‐6 secreted by PMN may play a regulatory role in the immune response by modulating the TNF‐α expression and its production by PBMC. This may have a significant influence on an early phase of the inflammation and other reactions mediated by TNF‐α.
Bloodstream Infection in Neutropenic Cancer Patients Related to Short-Term Nontunnelled Catheters Determined by Quantitative Blood Cultures, Differential Time to Positivity, and Molecular Epidemiological Typing with Pulsed-Field Gel Electrophoresis
Authors: Seifert Harald , Cornely Oliver , Seggewiss Kerstin , Decker Mathias , Stefanik Danuta , Wisplinghoff Hilmar , Fätkenheuer Gerd 
Published: January 2003

ABSTRACT
To determine the rate of catheter-related bloodstream infection (CRBSI) among cases of primary bloodstream infection (BSI) in febrile neutropenic cancer patients with short-term nontunnelled catheters, quantitative paired blood cultures (Isolator) from the central venous catheter (CVC) and peripheral vein were obtained between November 1999 and January 2001. Bactec blood culture bottles were obtained to determine the differential time to positivity (DTP). CRBSI was defined as a quantitative blood culture ratio of >5:1 (CVC versus peripheral) with proven identity of isolates from positive peripheral and CVC blood cultures as confirmed by pulsed-field gel electrophoresis. Forty-nine episodes of primary BSI were detected among 235 cancer patients with febrile neutropenia. Of these, 18 episodes (37%) were CRBSI and 31 (63%) were BSI with an unknown portal of entry. Coagulase-negative staphylococci were present in nine cases of CRBSI (50%). The identity of isolates from peripheral and CVC blood cultures was confirmed in all cases. Earlier positivity (>2 h) of CVC-drawn versus peripheral blood cultures was observed in 18 of 22 CRBSI-associated blood cultures (sensitivity, 82%; specificity, 88%; positive predictive value, 75%; negative predictive value, 92%). In summary, CRBSI accounted for 37% of cases of primary BSI in this population of neutropenic cancer patients. DTP compares favourably with quantitative blood cultures for the diagnosis of CRBSI and may be particularly useful for patients in whom catheter salvage is highly desirable.
Genetic Diversity of
Vibrio cholerae
O1 in Argentina and Emergence of a New Variant
Authors: Pichel Mariana , Rivas Marta , Chinen Isabel , Martín Fernando , Ibarra Cristina , Binsztein Norma 
Published: January 2003

ABSTRACT

The genetic diversity of
Vibrio cholerae
O1 strains from Argentina was estimated by random amplified polymorphic DNA (RAPD) analysis and pulsed-field gel electrophoresis (PFGE). Twenty-nine isolates carrying the virulence genes
ctxA
,
zot
,
ace
, and
tcpA
appeared to represent a single clone by both typing methods; while 11 strains lacking these virulence genes exhibited several heterogeneous RAPD and PFGE patterns. Among the last group, a set of isolates from the province Tucumán showed a single RAPD pattern and four closely related PFGE profiles. These strains, isolated from patients with diarrhea, did not produce the major
V. cholerae
O1 virulence determinants, yet cell supernatants of these isolates caused a heat-labile cytotoxic effect on Vero and Y-1 cells and elicited significant variations on the water flux and short-circuit current in human small intestine mounted in an Ussing chamber. All these effects were completely abolished by incubation with a specific antiserum against El Tor hemolysin, suggesting that this virulence factor was responsible for the toxic activity on both the epithelial cells and the small intestine specimens and may hence be involved in the development of diarrhea. We propose “Tucumán variant” as the designation for this new cluster of cholera toxin-negative
V. cholerae
O1 strains.
Evaluation of Two New Automated Assays for Hepatitis B Virus Surface Antigen (HBsAg) Detection: IMMULITE HBsAg and IMMULITE 2000 HBsAg
Authors: Weber Bernard , Dengler Thomas , Berger Annemarie , Doerr Hans Wilhelm , Rabenau Holger 
Published: January 2003

ABSTRACT
In recent years the diagnostic industry has developed new automated immunoassays for the qualitative detection of hepatitis B virus (HBV) surface antigen (HBsAg) in serum and plasma samples that are performed on analyzers that permit a high-speed throughput, random access, and primary tube sampling. The aim of the present study was the evaluation of two new automated HBsAg screening assays, IMMULITE HBsAg and IMMULITE 2000 HBsAg, from Diagnostic Products Corporation. The new HBsAg assays were compared to well-established tests (Auszyme Monoclonal [overnight incubation, version B], IMx HBsAg, AxSYM HBsAg, and Prism HBsAg [all from Abbott] and Elecsys HBsAg [Roche Diagnostics]). In the evaluation were included seroconversion panels, sera from the acute and chronic phases of infection, dilution series of various HBsAg standards, HBV subtypes and S gene mutants. To challenge the specificity of the new assays, sera from HBsAg-negative blood donors, pregnant women, and dialysis and hospitalized patients and potentially cross-reactive samples were investigated. IMMULITE HBsAg and IMMULITE 2000 HBsAg, although not as sensitive as the Elecsys HBsAg assay, were equivalent to the AxSYM HBsAg assay and showed a higher sensitivity than the Auszyme Monoclonal B and IMx HBsAg systems for detection of acute infection in seroconversion panels. The specificities (100%) of both IMMULITE assays on unselected blood donors and potentially interfering samples were comparable to those of the alternative assays after repeated testing. In conclusion, the new IMMULITE HBsAg and IMMULITE 2000 HBsAg assays show a good sensitivity for HBsAg detection compared to other well-established tests. The specificity on repeatedly tested samples was equivalent to that of the alternative assays. The rapid turnaround time, primary tube sampling, and on-board dilution make it an interesting assay system for clinical laboratory diagnosis.
Comparative Genotyping of
Campylobacter jejuni
by Amplified Fragment Length Polymorphism, Multilocus Sequence Typing, and Short Repeat Sequencing: Strain Diversity, Host Range, and Recombination
Authors: Schouls Leo M. , Reulen Sanne , Duim Birgitta , Wagenaar Jaap A. , Willems Rob J. L. , Dingle Kate E. , Colles Frances M. , Van Embden Jan D. A. 
Published: January 2003

ABSTRACT

Three molecular typing methods were used to study the relationships among 184
Campylobacter
strains isolated from humans, cattle, and chickens. All strains were genotyped by amplified fragment length polymorphism (AFLP) analysis, multilocus sequence typing (MLST), and sequence analysis of a genomic region with short tandem repeats designated clustered regularly interspaced short palindromic repeats (CRISPRs). MLST and AFLP analysis yielded more than 100 different profiles and patterns, respectively. These multiple-locus typing methods resulted in similar genetic clustering, indicating that both are useful in disclosing genetic relationships between
Campylobacter jejuni
isolates. Group separation analysis of the AFLP analysis and MLST data revealed an unexpected association between cattle and human strains, suggesting a common source of infection. Analysis of the polymorphic CRISPR region carrying short repeats allowed about two-thirds of the typeable strains to be distinguished, similar to AFLP analysis and MLST. The three methods proved to be equally powerful in identifying strains from outbreaks of human campylobacteriosis. Analysis of the MLST data showed that intra- and interspecies recombination occurs frequently and that the role of recombination in sequence variation is 50 times greater than that of mutation. Examination of strains cultured from cecum swabs revealed that individual chickens harbored multiple
Campylobacter
strain types and that some genotypes were found in more than one chicken. We conclude that typing of
Campylobacter
strains is useful for identification of outbreaks but is probably not useful for source tracing and global epidemiology because of carriage of strains of multiple types and an extremely high diversity of strains in animals.
Occurrence and Phenotypic Characteristics of Extended-Spectrum β-Lactamases among Members of the Family
Enterobacteriaceae
at the Tel-Aviv Medical Center (Israel) and Evaluation of Diagnostic Tests
Authors: Navon-Venezia Shiri , Hammer-Munz Orly , Schwartz David , Turner Dan , Kuzmenko Boris , Carmeli Yehuda 
Published: January 2003

ABSTRACT

We assessed the prevalence and phenotypic characteristics of extended-spectrum β-lactamase (ESBL) producers among cefuroxime-resistant (CXM-R) (MIC ≥ 32 μg/ml) members of the family
Enterobacteriaceae
in our institution. The 438 CXM-R clinical isolates obtained from nonurine sources among inpatients were screened. ESBL production was confirmed by disk diffusion assay using cefpodoxime (CPD), cefotaxime (CTX), and ceftazidime (CTZ) with and without clavulanate (CLAV). A difference of ≥5 mm in the size of the zone of inhibition in the presence of CLAV for at least one of the agents was considered representative of the ESBL phenotype: 186 isolates (42.5%) were confirmed as ESBL producers. The isolates tested and the rates of ESBL producers were as follows:
Klebsiella
spp. (
n
= 81), 79%;
Proteus
spp. (
n
= 58), 62%;
Escherichia coli
(
n
= 64), 53%;
Enterobacter
spp. (
n
= 69), 42%;
Serratia
spp. (
n
= 70), 14%;
Citrobacter
spp. (
n
= 25), 24%;
Providencia
spp. (
n
= 21), 24%;
Morganella
spp. (
n
= 41), 5%; and
Kluyvera
(
n
= 3), 0%. The overall sensitivity of isolated ESBL confirmatory tests was 79% for CPD-CLAV, 66% for CTZ-CLAV, and 91% for CTX-CLAV. Sensitivities of CTZ-CLAV confirmatory tests for
Klebsiella
spp.,
Proteus
spp.,
E. coli
, and
Enterobacter
spp. were 84, 22, 76, and 62%, respectively, and those for CTX-CLAV were 95, 97, 94, and 83%, respectively. They were 90% for CPD-CLAV and CTZ-CLAV, 95% for CPD-CLAV and CTX-CLAV, and 100% for CTZ-CLAV and CTX-CLAV. ESBL production was highly prevalent among
Enterobacteriaceae.
Using resistance to CXM as an ESBL screening criterion is a suitable option in high-incidence areas where
Klebsiella
spp. are not the dominant ESBL producers. This screening criterion may simplify the screening test and improve its sensitivity, although at the price of testing more isolates. The CTX-CLAV combination confirmed ESBL producers better than the CTZ-CLAV combination, with sensitivity varying between species. Combined CTZ-CLAV and CTX-CLAV testing detected all these strains; CPD-CLAV provided no additional benefit.
Frequent Recovery of a Single Clonal Type of Multidrug-Resistant
Staphylococcus aureus
from Patients in Two Hospitals in Taiwan and China
Authors: Aires de Sousa M. , Crisóstomo M. I. , Santos Sanches I. , Wu J. S. , Fuzhong J. , Tomasz A. , de Lencastre H. 
Published: January 2003

ABSTRACT

One hundred thirty-two methicillin-resistant
Staphylococcus aureus
(MRSA) isolates recovered from patients with
S. aureus
infections between January 1998 and February 1999 in two hospitals, one located in Taipei, Taiwan, and another in Nanjing, People's Republic of China, were examined for antibiotic susceptibility and for clonal type by a combination of three methods: hybridization of
Cla
I restriction digests with
mecA
- and Tn
554
-specific DNA probes and pulsed-field gel electrophoresis of chromosomal
Sma
I digests. Selected isolates representing each clonal type were also analyzed by
spaA
typing, multilocus sequence typing, and a multiplex PCR method capable of identifying the structural type of the staphylococcal cassette chromosome
mec
(SCC
mec
) carried by the bacteria. The overwhelming majority of isolates (126 of 132 or 95%) belonged to minor variants of a single clonal type resembling the Brazilian and Hungarian epidemic MRSA clones, which showed a common
spaA
type and which were either sequence type 239 (ST239) or ST241 (a single-locus variant of ST239) in association with SCC
mec
type III or IIIA.
Multicenter Evaluation of the Performance Characteristics of the NucliSens HIV-1 QT Assay Used for Quantitation of Human Immunodeficiency Virus Type 1 RNA
Authors: Ginocchio Christine C. , Kemper Marti , Stellrecht Kathleen A. , Witt Donald J. 
Published: January 2003

ABSTRACT

The analytical performance of the NucliSens HIV-1 QT assay, a highly sensitive test based on nucleic acid sequence-based amplification technology, was evaluated in a multicenter trial. Assay specificity was evaluated with 502 plasma (EDTA) specimens from human immunodeficiency virus type 1 (HIV-1)-seronegative volunteer donors. No HIV-1 RNA was reported in any of the donor specimens. Analytical sensitivity and reproducibility were estimated with panels prepared from a high-titer well-characterized HIV-1 RNA stock (5.84 × 10
8
RNA copies/ml). The assay's dynamic range was linear from 10
6
to 10
1
HIV-1 RNA copies, with a lower detectable limit of 25 copies/ml and a 95% detection rate of 176 copies/ml. Sensitivity of the assay to detect HIV-1 RNA in clinical specimens from patients (
n
= 101) and in commercially available or prepared panels (
n
= 24) was compared with NASBA HIV-1 RNA QT (an earlier version of NucliSens HIV-1 QT) and with the Food and Drug Administration-approved standard and ultrasensitive AMPLICOR HIV-1 MONITOR, version 1.0, assays. Detection of HIV-1 RNA was reproducible over a 5-log range (mean standard deviation = 0.15 log). The NucliSens and the standard AMPLICOR assays were equivalent in detection of HIV-1 RNA (concentration, 10
3
to 10
5
copies/ml) in 57 clinical specimens. The NucliSens assay was more sensitive in detecting HIV-1 RNA at lower concentrations (≤10
2
copies/ml) (44 of 44) than either the standard AMPLICOR test (12 of 19) or the NASBA assay (10 of 25). A 25% increase in HIV-1 RNA detection frequency with panels was observed with the NucliSens assay (23 of 24) compared with the standard AMPLICOR test (17 of 24). The new assay was highly specific and demonstrated good sensitivity with a broad linear dynamic range.
Pneumocystis jiroveci
Internal Transcribed Spacer Types in Patients Colonized by the Fungus and in Patients with Pneumocystosis from the Same French Geographic Region
Authors: Nevez Gilles , Totet Anne , Jounieaux Vincent , Schmit Jean-Luc , Dei-Cas Eduardo , Raccurt Christian 
Published: January 2003

ABSTRACT

Pneumocystis jiroveci
(human-derived
Pneumocystis
) infections can display a broad spectrum of clinical presentations, of which pulmonary colonization with the fungus may represent an important part, occurring frequently in patients with various underlying diseases and presenting alternative diagnoses of acute pneumocystosis (
Pneumocystis carinii
pneumonia [PCP]). There are few data concerning the
P. jiroveci
genotypes involved in pulmonary colonization, whereas several genotypes responsible for PCP in immunocompromised patients have been described. In this study,
P. jiroveci
genotypes have retrospectively been investigated and compared in 6 colonized patients and in 11 patients with PCP who were in the same hospital. Seventeen archival bronchoalveolar lavage samples were genotyped at internal-transcribed spacer 1 (ITS1) and ITS2 of the nuclear rRNA operon. Fourteen different genotypes were identified, of which 1 was found only in colonized patients, 10 were found only in patients with PCP, and 3 were found in both patient populations. Mixed infections were diagnosed in 2 of the 6 colonized patients and in 6 of the 11 patients with PCP. The results show that similar genotypes can be responsible for PCP as well as pulmonary colonization. There is a high diversity of genotypes in colonized patients and in patients with PCP. Mixed infections may occur in these two patient populations. These shared features of
P. jiroveci
ITS genotypes in colonized patients and patients with PCP suggest that human populations infected by
P. jiroveci
, whatever the clinical manifestation, may play a role as a common reservoir for the fungus.
Characteristics of
Massilia timonae
and
Massilia timonae
-Like Isolates from Human Patients, with an Emended Description of the Species
Authors: Lindquist David , Murrill Donna , Burran W. Patrick , Winans Gloria , Janda J. Michael , Probert Will 
Published: January 2003

ABSTRACT

The description of
Massilia timonae
, a nonfermentative aerobic gram-negative rod, was based on a single strain. A subsequent report of a second isolate has been recently published. Phenotypic descriptions of these two strains were based primarily on commercial test kit results. We have identified three additional strains as
M. timonae
by 16S rRNA sequence analysis and have characterized them phenotypically in parallel with the type strain of
M. timonae
, CIP 105350, by conventional test methods. A fourth strain, designated
M. timonae
-like, was also characterized. All four strains were isolated from human patients: two were blood isolates, one was isolated from cerebrospinal fluid, and one was isolated from bone. The four strains and the type strain were quite similar phenotypically. However, in contrast to the original description, the strains were found to be oxidase positive and arginine dihydrolase negative and to have lateral flagella as well as a single polar flagellum. Additionally the strains produced acid oxidatively from some carbohydrates. Other phenotypic characteristics, including cellular fatty acids, agreed with the original description. Based on our emended description,
M. timonae
and
M. timonae
-like strains can be differentiated from other aerobic nonfermentative gram-negative rods by conventional biochemical tests combined with cellular fatty acid analysis.