bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effect of Hypoxia on Urocortin Production in Human Gestational Trophoblasts In Vitro
Authors: Choy Mei Y. , Leung Tse N. , Leung Po S. , Lau Tze K. 
Published: July 2004

Problem:  Urocortin is produced by the placenta throughout pregnancy but its regulation remains unknown. The effect of hypoxia on placental urocortin production is not known. The aim of this study was to determine the effect of in vitro hypoxia on human trophoblastic urocortin production.Method of study:  Placental explants and primary cultures were incubated in anaerobe hypoxic bags for 24 h in a humidified incubator. Urocortin peptide secretion and mRNA (messenger RNA) production was determined by enzyme‐linked immunosorbent assay and reverse transcription‐polymerase chain reaction, respectively. Morphological and functional integrity was verified by immunohistochemical analysis of urocortin expression. Vascular endothelial growth factor expression was used to verify the generation of cellular hypoxia in our in vitro system.Results:  Hypoxia did not affect urocortin secretion or mRNA expression in explant and single‐cell cultures. Production was greater from first trimester than term explants and from single‐cell primary cultures more than from explant cultures.Conclusions:  Hypoxia does not influence human placental urocortin secretion or mRNA expression in vitro.
Endometrial and Peritoneal Macrophages: Expression of Activation and Adhesion Molecules
Authors: Eidukaite Audrone , Tamosiunas Vytas 
Published: July 2004

Problem:  Macrophages are highly individualized in tissues and their activities are a reflection of systemic and local environmental signals. The expression of activation (CD69, CD71) and adhesion (CD54) molecules on the surface of CD14+ endometrial macrophages at various phases of the menstrual cycle was compared with the cell surface receptors of peritoneal fluid macrophages.Method of study:  Two‐colour‐flow cytometry was used to determine the peritoneal and endometrial macrophage phenotype.Results:  Endometrium macrophages expressed a lower level of CD69+ and CD54+ macrophages than peritoneal macrophages. However, CD71 receptors displayed similar expression in both macrophage populations, endometrium and peritoneal, except during the proliferative phase.Conclusion:  These findings demonstrate the differences between macrophages from endometrium and peritoneal fluid with regard to CD69, CD71 and CD54 expression. In addition, increased numbers of endometrial macrophages in the late secretory phase of the menstrual cycle suggest that they may play a role in menstruation.
Relationship between Cervical Mucus Interleukin‐8 Concentrations and Vaginal Bacteria in Pregnancy
Authors: Sakai Masatoshi , Ishiyama Akihiko , Tabata Mika , Sasaki Yasushi , Yoneda Satoshi , Shiozaki Arihiro , Saito Shigeru 
Published: July 2004

Problem:  High interleukin (IL)‐8 concentration in cervical mucus in the second trimester is a risk factor for premature birth. We investigated the relationship between vaginal pathogens and IL‐8 in cervical mucus.Method of study:  In 501 women with single pregnancy, vaginal secretions were cultured for bacteria and cervical mucus IL‐8 concentrations were measured between 20 and 24 gestational weeks.Results:  Lactobacillus species were detected in 56.0% of 84 subjects with high IL‐8 (≥377 ng/mL), significantly less often than in 417 subjects with IL‐8 below 377 ng/mL (84.7%; P < 0.0001). Anaerobic pathogens were detected in 83.3% of high IL‐8 subjects, significantly more often than in normal IL‐8 subjects (43.9%; P < 0.0001). By multivariate analysis, cervical IL‐8 was significantly high only in subjects without Lactobacillus species; they showed a significantly higher prematurity rate than Lactobacillus‐positive subjects.Conclusions:  Absence of vaginal Lactobacilli was associated with increased cervical IL‐8 and increased risk of premature delivery.
Leukemia Inhibitory Factor: An Important Regulator of Endometrial Function
Authors: Kondera‐Anasz Zdzisława , Sikora Justyna , Mielczarek‐Palacz Aleksandra 
Published: July 2004

Problem:  Leukemia inhibitory factor (LIF) is multifunctional cytokine that displays biological activities in different cells, including endometrial cells. The aim of this study is to describe implications of LIF on a physiological function of endometrium.Method of study:  The role of LIF in the endometrial function is reviewed and summarized from the available literature.Results:  LIF plays an important role in a physiological function of endometrium. In human endometrial LIF expression depends on cellular localizations, steroid hormones, menstrual stages and a local cytokine network. Stronger LIF expression exists in an endometrial epithelium during a luteal phase of the menstrual cycle, which coincides with the time of an implantation. The impairments of the endometrial LIF expression may play a significant role in the pathological processes involving implantation and the infertility.Conclusions:  There is a substantial evidence that LIF is a potential regulator of the endometrial function and might be one of the factors that play a key role in human reproduction.
Analysis of Clonal Composition of
Mycobacterium tuberculosis
Isolates in Primary Infections in Children
Authors: García de Viedma Darío , Marín Mercedes , Ruiz María Jesús , Bouza Emilio 
Published: August 2004

ABSTRACT

The assumption that
Mycobacterium tuberculosis
infections should be considered clonally homogeneous has been weakened in the last few years. Recent studies have shown (i) the isolation of different
M. tuberculosis
strains from sequential episodes, (ii) mixed infections by two
M. tuberculosis
strains, and (iii) genetic variations in
M. tuberculosis
subpopulations due to microevolution events. Nevertheless, it is unknown whether clonal heterogeneity could be found in the initial steps of
M. tuberculosis
infection, i.e., the primary infection. In the present study we analyzed the clonal composition of the
M. tuberculosis
isolates causing primary infections in children. Cultures were clonally homogeneous in most cases (11 of 12). In 1 of the 12 cases (8.3%), clonal heterogeneity among the
M. tuberculosis
isolates was found by spoligotyping and IS
6110
-based restriction fragment length polymorphism analysis. This case occurred in a 2-year-old child in whom microevolution events were unlikely and who had no risk factors for overexposure to
M. tuberculosis
. Clonal heterogeneity should also be considered in primary
M. tuberculosis
infections, including circumstances in which it is usually unexpected.
Sex Pheromone Response, Clumping, and Slime Production in Enterococcal Strains Isolated from Occluded Biliary Stents
Authors: Donelli Gianfranco , Paoletti Claudia , Baldassarri Lucilla , Guaglianone Emilio , Di Rosa Roberta , Magi Gloria , Spinaci Cinzia , Facinelli Bruna 
Published: August 2004

ABSTRACT

Bile-resistant bacteria, particularly gram-positive
Enterococcus faecalis
and
Enterococcus faecium
, play an important role in biliary stent occlusion, because their sessile mode of growth protects them against host defenses and antimicrobial agents. Twelve
E. faecalis
and seven
E. faecium
strains isolated from occluded biliary stents have been investigated for slime production, presence of aggregation substance genes, and ability to adhere to Caco-2 cells. Ten isolates were strong producers of slime, and seven isolates produced clumps when exposed to pheromones of
E. faecalis
JH2-2 and/or OG1RF. The small
E. faecium
clumps differed from the large clumps of
E. faecalis
and were similar to those of
E. faecium
LS10(pBRG1) carrying a pheromone response plasmid. After induction with pheromones, the adhesion to Caco-2 cells of clumping-positive strains was found to increase from two- to fourfold. Amplicons of the expected size were detected in three clumping-positive and three clumping-negative
E. faecalis
isolates by using primers (
agg
) internal to a highly conserved region of the
E. faecalis
pheromone response plasmids pAD1, pPD1, and pCF10 and primers internal to
prgB
of the
E. faecalis
plasmid pCF10. The
agg
/
prgB
-positive
E. faecalis
strains were also positive in Southern hybridization experiments with a
prgB
-specific probe. No PCR products were obtained with the same primers from four clumping-positive isolates (one
E. faecalis
and three
E. faecium
strains), which were also Southern hybridization negative. Our results demonstrate that slime production and pheromone response are both present in isolated enterococci, suggesting that clinical strains with these features might have a selective advantage in colonizing biliary stents.
Suicide PCR on Skin Biopsy Specimens for Diagnosis of Rickettsioses
Authors: Fournier Pierre-Edouard , Raoult Didier 
Published: August 2004

ABSTRACTAs rickettsioses may be severe diseases andRickettsia prowazekiiis a potential agent of bioterrorism, highly efficient diagnostic techniques are required to detect rickettsiae in patients. We developed a nested PCR assay using single-use primers targeting single-use gene fragments present in the genomes of bothRickettsia conoriiandR. prowazekii. We used this “suicide” PCR with DNA from 103 skin biopsy specimens from patients who definitely had a rickettiosis, 109 skin biopsy specimens from patients who possibly had a rickettsiosis, and 50 skin biopsy specimens from patients with nonrickettsial diseases. The suicide PCR detected “R. conorii conorii” in 38 biopsy specimens,R. africaein 28 biopsy specimens,R. slovacain 12 biopsy specimens, “R. sibirica mongolotimonae” in 5 biopsy specimens,R. aeschlimanniiin 2 biopsy specimens, and “R. conorii caspia” and “R. sibirica sibirica” in 1 biopsy specimen each. The technique had a specificity of 100% and a sensitivity of 68%. It was 2.2 times more sensitive than culture (P< 10−2) and 1.5 times more sensitive than regular PCR (P< 10−2). The efficacy of the suicide PCR was reduced by antibiotic therapy prior to biopsy (P< 10−2) and was increased when it was performed with eschar biopsy specimens (P= 0.03). We propose the use of the suicide PCR as a sensitive, specific, and versatile technique for improving the diagnosis of rickettsioses, especially when it is used on eschar biopsy specimens taken prior to antibiotic therapy.
Characterization of a Trinucleotide Repeat Sequence (CGG)
5
and Potential Use in Restriction Fragment Length Polymorphism Typing of
Mycobacterium tuberculosis
Authors: Otsuka Yayoi , Parniewski Pawel , Zwolska Zofia , Kai Masatake , Fujino Tomoko , Kirikae Fumiko , Toyota Emiko , Kudo Koichiro , Kuratsuji Tadatoshi , Kirikae Teruo 
Published: August 2004

ABSTRACT

The genomes of 28 bacterial strains, including mycobacterial species
Mycobacterium tuberculosis
and
Mycobacterium bovis
, were analyzed for the presence of a special class of microsatellite, that of trinucleotide repeat sequences (TRS). Results of a search of all 10 possible TRS motifs (i.e., CCT, CGG, CTG, GAA, GAT, GTA, GTC, GTG, GTT, and TAT) with five or more repeating units showed that (CGG)
5
was highly represented within the genomic DNA of
M. tuberculosis
and
M. bovis
. Most of the (CGG)
5
repeats in the genome were within the open reading frames of two large gene families encoding PE_PGRS and PPE proteins that have the motifs Pro-Glu (PE) and Pro-Pro-Glu (PPE). (CGG)
5
-probed Southern hybridization showed that some mycobacterial species, such as
Mycobacterium marinum
,
Mycobacterium kansasii
, and
Mycobacterium szulgai
, possess many copies of (CGG)
5
in their genomes. Analysis of clinical isolates obtained from Tokyo and Warsaw with both IS
6110
and (CGG)
5
probes showed that there is an association between the fingerprinting patterns and the geographic origin of the isolates and that (CGG)
5
fingerprinting patterns were relatively more stable than IS
6110
patterns. The (CGG)
5
repeat is a unique sequence for some mycobacterial species, and (CGG)
5
fingerprinting can be used as an epidemiologic method for these species as well as IS
6110
fingerprinting can. If these two fingerprinting methods are used together, the precise analysis of
M. tuberculosis
isolates will be accomplished. (CGG)
5
-based fingerprinting is particularly useful for
M. tuberculosis
isolates with few or no insertion elements and for the identification of other mycobacterial species when informative probes are lacking.
Characterization and Prevalence of MefA, MefE, and the Associated
msr
(D) Gene in
Streptococcus pneumoniae
Clinical Isolates
Authors: Daly Melissa M. , Doktor Stella , Flamm Robert , Shortridge Dee 
Published: August 2004

ABSTRACT

Recent work has shown that the efflux genes in
Streptococcus pneumoniae
that are responsible for acquired macrolide resistance can be distinguished as either
mef
(E) or
mef
(A). The genetic elements on which
mef
(A) and
mef
(E) are found also carry an open reading frame (ORF) that is 56% homologous to
msr
(A) in
Staphylococcus
. The prevalence of
mef
(A/E) and of the
msr
-like ORF [
msr
(D)] was evaluated in 153

mef
+
S. pneumoniae

clinical isolates collected in North America, Europe, Africa, and Asia from 1997 to 2002. Clinical isolates were screened with PCR primers specific for either
mef
(A) or
mef
(E) and for
msr
(D).
mef
(A),
mef
(E), and
msr
(D) were cloned from

mef
+

strains and transformed into a susceptible, competent strain of
S. pneumoniae
. The transformants were tested for antimicrobial susceptibilities and efflux pump induction. The results of this work demonstrated that
mef
(A) is more often isolated in parts of Europe, with some incidence in Canada, and that the
msr
-like gene alone can confer the efflux phenotype.
Comparison of Results of Fluconazole Disk Diffusion Testing for
Candida
Species with Results from a Central Reference Laboratory in the ARTEMIS Global Antifungal Surveillance Program
Authors: Pfaller M. A. , Hazen K. C. , Messer S. A. , Boyken L. , Tendolkar S. , Hollis R. J. , Diekema D. J. 
Published: August 2004

ABSTRACT

The accuracy of antifungal susceptibility tests is important for accurate resistance surveillance and for the clinical management of patients with serious infections. Our main objective was to compare the results of fluconazole disk diffusion testing of
Candida
spp. performed by ARTEMIS participating centers with disk diffusion and MIC results obtained by the central reference laboratory. A total of 2,949 isolates of
Candida
spp. were tested by NCCLS disk diffusion and reference broth microdilution methods in the central reference laboratory. These results were compared to the results of disk diffusion testing performed in the 54 participating centers. All tests were performed and interpreted following NCCLS recommendations. Overall categorical agreement between participant disk diffusion test results and reference laboratory MIC results was 87.4%, with 0.2% very major errors (VME) and 3.3% major errors (ME). The categorical agreement between the disk diffusion test results obtained in the reference laboratory with the MIC test results was similar: 92.8%. Likewise, good agreement was observed between participant disk diffusion test results and reference laboratory disk diffusion test results: 90.4%, 0.4% VME, and 3.4% ME. The disk diffusion test was especially reliable in detecting those isolates of
Candida
spp. that were characterized as resistant by reference MIC testing. External quality assurance data obtained by surveillance programs such as the ARTEMIS Global Antifungal Surveillance Program ensure the generation of useful surveillance data and result in the continued improvement of antifungal susceptibility testing practices.
Development of Two PCR-Based Techniques for Detecting Helical and Coccoid Forms of
Helicobacter pylori
Authors: Shahamat M. , Alavi M. , Watts J. E. M. , Gonzalez J. M. , Sowers K. R. , Maeder D. W. , Robb F. T. 
Published: August 2004

ABSTRACT

The primary mode of transmission of
Helicobacter pylori
, a human pathogen carried by more than half the population worldwide, is still unresolved. Some epidemiological data suggest water as a possible transmission route.
H. pylori
in the environment transforms into a nonculturable, coccoid form, which frequently results in the failure to detect this bacterium in environmental samples by conventional culture techniques. To overcome limitations associated with culturing, molecular approaches based on DNA amplification by PCR have been developed and used for the detection of
H. pylori
in clinical and environmental samples. Our results showed the
glmM
gene as the most promising target for detection of
H. pylori
by PCR amplification. Under optimal amplification conditions,
glmM
-specific primers generated PCR-amplified products that were specific for
H. pylori
and some other
Helicobacter
species. Genome sequence analysis revealed the existence of a conserved region linked to a hypervariable region upstream of the 16S rRNA gene of
H. pylori
. Selective PCR primer sets targeting this sequence were evaluated for the specific detection of
H. pylori
. One primer set, Cluster2 and B1J99, were shown to be highly specific for
H. pylori
strains and did not produce any PCR products when other
Helicobacter
species and other bacterial species were analyzed. In tests with 32 strains of
H. pylori
, 6 strains of other
Helicobacter
species, 8 strains of
Campylobacter jejuni
, and 21 strains belonging to different genera, the primers for
glmM
were selective for the
Helicobacter
genus and the primers containing the region flanking the 16S rRNA gene were selective for
H. pylori
species only. The combination of two sensitive PCR-based methods, one targeting the
glmM
gene and the other targeting a hypervariable flanking region upstream of the 16S rRNA gene, are complementary to each other. Whereas the
glmM
-specific primers provide a rapid, sensitive presumptive assay for the presence of
H. pylori
and closely related
Helicobacter
spp., the primers for sequences flanking the 16S rRNA gene can confirm the presence of
H. pylori
and locate the potential source of this bacterium.
Rapid Identification of
Nocardia farcinica
Clinical Isolates by a PCR Assay Targeting a 314-Base-Pair Species-Specific DNA Fragment
Authors: Brown June M. , Pham Kim N. , McNeil Michael M. , Lasker Brent A. 
Published: August 2004

ABSTRACT

Nocardia farcinica
is the most clinically significant species within the
Nocardia asteroides
complex. Differentiation of
N. farcinica
from other members of
N. asteroides
complex is important because this species characteristically demonstrates resistance to several extended-spectrum antimicrobial agents. Traditional phenotypic characterization of this species is time- and labor-intensive and often leads to misidentification in the clinical microbiology laboratory. We previously observed a 409-bp product for all strains of
N. farcinica
by using randomly amplified polymorphic DNA analysis with the primer DKU49. In this investigation, the 409-bp fragment was sequenced and then used to design a specific primer pair, Nf1 (16-mer) and Nf2 (16-mer), complementary to the 409-bp fragment. PCR amplification of genomic DNA from 28
N. farcinica
isolates with Nf1 and Nf2 generated a single intense 314-bp fragment. The specificity of the assay with these primers was verified, since there were no PCR amplification products observed from heterologous nocardial species (
n
= 59) or other related bacterial genera (
n
= 41). Restriction enzyme digestion using CfoI and direct sequencing of the 314-bp fragment further confirmed the specificity of the assay for
N. farcinica
. This highly sensitive and specific PCR assay provides a rapid (within 1 day of obtaining DNA) method for identification of this medically important emerging pathogen. Rapid diagnosis of
N. farcinica
infection may allow for earlier initiation of effective therapy, thus improving patient outcome.
Comparison of the Directigen Flu A+B Membrane Enzyme Immunoassay with Viral Culture for Rapid Detection of Influenza A and B Viruses in Respiratory Specimens
Authors: Cazacu Andreea C. , Chung Sooyoung E. , Greer Jewel , Demmler Gail J. 
Published: August 2004

ABSTRACT
The performance of a commercially available, rapid membrane enzyme immunoassay for influenza A and B virus detection was compared to that of viral culture in 4,092 respiratory specimens collected from patients presenting with respiratory symptoms during the 2002-2003 influenza season. The test's overall sensitivity was 43.83%, lower than previously reported but similar for detection of both influenza A and B viruses (42.98 versus 44.76%). However, specificity, 99.74%, was excellent for both influenza A and B viruses (99.82 versus 99.92%). These values make this test a very good confirmatory test when clinical suspicion is high, but a less accurate screening test for large populations.
Concurrent Infections with Vector-Borne Pathogens Associated with Fatal Hemolytic Anemia in a Cattle Herd in Switzerland
Authors: Hofmann-Lehmann Regina , Meli Marina L. , Dreher Ute M. , Gönczi Enikö , Deplazes Peter , Braun Ueli , Engels Monika , Schüpbach Jörg , Jörger Kaspar , Thoma Rudolf , Griot Christian , Stärk Katharina D. C. , Willi Barbara , Schmidt Joseph , Kocan Katherine M. , Lutz Hans 
Published: August 2004

ABSTRACTBovine anaplasmosis is a vector-borne disease that results in substantial economic losses in other parts of the world but so far not in northern Europe. In August 2002, a fatal disease outbreak was reported in a large dairy herd in the Swiss canton of Grisons. Diseased animals experienced fever, anorexia, agalactia, and depression. Anemia, ectoparasite infestation, and, occasionally, hemoglobinuria were observed. To determine the roles of vector-borne pathogens and to characterize the disease, blood samples were collected from all 286 animals: 50% of the cows were anemic. Upon microscopic examination of red blood cells,Anaplasma marginaleinclusion bodies were found in 47% of the cows. The infection was confirmed serologically and by molecular methods. Interestingly, we also found evidence of infections withAnaplasma phagocytophilum, largeBabesiaandTheileriaspp., andMycoplasma wenyonii. The last two species had not previously been described in Switzerland. Anemia was significantly associated with the presence of the infectious agents detected, with the exception ofA. phagocytophilum. Remarkably, concurrent infections with up to five infectious vector-borne agents were detected in 90% of the ill animals tested by PCR. We concluded thatA. marginalewas the major cause of the hemolytic anemia, while coinfections with other agents exacerbated the disease. This was the first severe disease outbreak associated with concurrent infections with vector-borne pathogens in alpine Switzerland; it was presumably curtailed by culling of the entire herd. It remains to be seen whether similar disease outbreaks will have to be anticipated in northern Europe in the future.
In Vivo Acquisition of High-Level Resistance to Imipenem inEscherichia coli
Authors: Poirel Laurent , Héritier Claire , Spicq Colette , Nordmann Patrice 
Published: August 2004

ABSTRACTFour clonally relatedEscherichia colistrains were isolated successively from bile duct of a girl suffering from sclerosing cholangitis. One of them, selected after an imipenem-containing regimen, was resistant to carbapenems and to broad-spectrum cephalosporins due to a plasmid-mediated cephalosporinase, CMY-2, and the lack of outer membrane proteins OmpF and OmpC.
Persistence of Human Immunodeficiency Virus Type 1 Subtype B DNA in Dried-Blood Samples on FTA Filter Paper
Authors: Li Chung-Chen , Beck Ingrid A. , Seidel Kristy D. , Frenkel Lisa M. 
Published: August 2004

ABSTRACT

The stability of human immunodeficiency virus type 1 (HIV-1) DNA in whole blood collected on filter paper (FTA Card) was evaluated. After >4 years of storage at room temperature in the dark our qualitative assay detected virus at a rate similar to that of our initial test (58 of 60, 97%;
P
= 0.16), suggesting long-term HIV-1 DNA stability.