bluuebunny/crossref_metadata_2025_split huggingface dataset:
Effects of pulsatile secretion of growth hormone (GH) on fat deposition in human GH transgenic rats
Authors: Furuhata Yasufumi , Nishihara Masugi , Takahashi Michio 
Published: April 2003

AbstractGrowth hormone (GH) is an endocrine regulator of glucose and lipid metabolism as well as body growth. GH levels are decreased and a unique pulsatile secretory pattern becomes obvious after puberty particularly in males. Coincidentally with this, males tend to deposit body fat. Experimental and clinical evidence has accumulated that obesity is associated with a decrease in GH levels. A strain of transgenic rats has been generated with severe obesity but normal nose-to-tail length, which has low circulating GH levels without pulsatility (human growth hormone (hGH) transgenic rats). The present review mainly focuses on recent and current work analysing the relationship between the occurrence of obesity and low GH levels and/or the absence of GH pulsatility in this transgenic animal model. This model has elevated blood glucose, non-esterified fatty acid, insulin and leptin levels associated with hyperphagia, suggesting that these rats also carry insulin- and leptin-resistant characteristics. hGH transgenic rats were subjected to a pair-feeding treatment to normalize food intake and chronic GH replacement to normalize GH levels. While the pair-feeding for 8 weeks successfully suppressed body-weight gain, the fat pad : body weight ratio remained very similar to freely-eating control hGH transgenic rats, which indicates the hyperphagia is not the sole contributor to the excess fat accumulation in this model. However, continuous elevation of peripheral hGH levels (approximately 2-fold) for 8 weeks by means of a slow-release vehicle resulted in a significant decrease in the fat mass : body weight ratios by 30 %. This GH treatment altered neither food intake nor body-weight gain. Thus, two characteristic phenotypes observed in the hGH transgenic rats, hyperphagia and obesity, seem to be closely related to GH levels and GH secretory pattern. This relationship might be working in the regulation of changes in seasonal body composition in wild animals.
Method for Quantitative Detection and Presumptive Identification of Group B Streptococci on Primary Plating
Authors: Hansen Søren Mose , Sørensen Uffe B. Skov 
Published: April 2003

ABSTRACT
Maternal prenatal screening for group B streptococci (GBS) followed by offering of intrapartum chemoprophylaxis to carriers is one of the strategies used to reduce the incidence of neonatal early-onset GBS infections. Culturing of vaginal and anorectal swab specimens in selective broth is the screening procedure recommended by the Centers for Disease Control and Prevention. This technique is sensitive; it does not, however, allow either evaluation of the degree of colonization or detection of cocolonization with several GBS clones. We have examined the carriage rate and population dynamics of GBS in a group of Danish women during pregnancy and 1 year after delivery using a new detection method. In the present paper we describe a mixed blood agar medium (MB agar) that identifies GBS in the primary cultures by detection of a double hemolysis pattern consisting of characteristic, large zones of partial hemolysis (“CAMP zones”) and of narrow zones of complete hemolysis. The MB agar was at least as sensitive as culturing in selective broth for detection of GBS in vaginal and anorectal swab specimens, and GBS strains could be identified directly on the primary plate due to the CAMP zones without the need for subculturing. The carriage rate of GBS in a group of Danish women was found to be more than 30%, a figure considerably higher than the rate that was reported previously.
Development of a Real-Time PCR Assay for Quantitative Detection of
Encephalitozoon intestinalis
DNA
Authors: Menotti Jean , Cassinat Bruno , Sarfati Claudine , Liguory Olivier , Derouin Francis , Molina Jean-Michel 
Published: April 2003

ABSTRACT

A new real-time PCR assay for quantitation of
Encephalitozoon intestinalis
DNA was developed which used a
Taq
Man fluorescent probe for specific detection. Serial dilutions of
E. intestinalis
spore suspensions obtained from tissue culture were used as external standards. The detection limit of the technique was 20 spores per ml, with a good interassay reproducibility (coefficient of variation of 7.1% for the suspension containing 20 spores/ml, 5.0% for the suspension containing 75 spores/ml and below 3.5% for higher concentrations). Quantitative detection of
E. intestinalis
DNA was similar whether the serial dilutions of spores were made in distilled water or in a stool suspension, allowing the use of the assay for stool specimens. The assay was then applied to 14 clinical specimens from 8 immunocompromised patients with proven
E. intestinalis
infection. The quantitation of the parasitic burden was achieved in stools, blood, urine, tissue biopsies, and bronchopulmonary specimens. The highest parasitic burdens were noted in stools, urine, and bronchopulmonary specimens, reaching 10
5
to 10
6
spores/g or ml. Dissemination of the infection was also evidenced in some patients by demonstration of
E. intestinalis
DNA in blood and serum. We conclude that real-time PCR is a valuable tool for quantitation of
E. intestinalis
burden in clinical specimens.
International Collaborative Study To Compare Reverse Transcriptase PCR Assays for Detection and Genotyping of Noroviruses
Authors: Vinjé Jan , Vennema Harry , Maunula Leena , von Bonsdorff Carl-Henrik , Hoehne Marina , Schreier Eckart , Richards Alison , Green Jon , Brown David , Beard Suzanne S. , Monroe Stephan S. , de Bruin Erwin , Svensson Lennart , Koopmans Marion P. G. 
Published: April 2003

ABSTRACTTo allow more rapid and internationally standardized assessment of the spread of noroviruses (previously called Norwalk-like viruses [NLVs]) as important food-borne pathogens, harmonization of methods for their detection is needed. Diagnosis of NLVs in clinical diagnostic laboratories is usually performed by reverse transciptase PCR (RT-PCR) assays. In the present study, the performance of five different RT-PCR assays for the detection of NLVs was evaluated in an international collaborative study by five laboratories in five countries with a coded panel of 91 fecal specimens. The assays were tested for their sensitivity, detection limit, and ease of standardization. In total, NLVs could be detected by at least one RT-PCR assay in 69 (84%) of the samples that originally tested positive. Sensitivity ranged from 52 to 73% overall and from 54 to 100% and 58 to 85% for genogroup I and II viruses, respectively. In all, 64% of the false-negative results were obtained with a set of diluted stools (n= 20) that may have lost quality upon storage. Sensitivity was improved when these samples were excluded from analysis. No one single assay stood out as the best, although the p1 assay demonstrated the most satisfactory overall performance. To promote comparability of data, this assay will be recommended for newly starting groups in future collaborative studies.
Activities of Fluconazole and Voriconazole against 1,586 Recent Clinical Isolates of
Candida
Species Determined by Broth Microdilution, Disk Diffusion, and Etest Methods: Report from The ARTEMIS Global Antifungal Susceptibility Program, 2001
Authors: Pfaller M. A. , Diekema D. J. , Messer S. A. , Boyken L. , Hollis R. J. 
Published: April 2003

ABSTRACT

The ARTEMIS Global Antifungal Susceptibility Program (ARTEMIS Program) was initiated in 2001 to provide focused surveillance of the activities of fluconazole and voriconazole against
Candida
spp. isolated from blood and other normally sterile sites. A total of 1,586 episodes of infection were detected at 61 international study sites. Overall, 57.7% of the infections were due to
Candida albicans
, followed by
C. glabrata
(14.8%),
C. parapsilosis
(12.5%),
C. tropicalis
(9.4%),
C. krusei
(2.7%), and
C. lusitaniae
(1.5%). Isolates of
C. albicans
,
C. parapsilosis
, and
C. tropicalis
were all highly susceptible to fluconazole (for 99% of the isolates the MICs were ≤8 μg/ml). Likewise, 99 to 100% of these species were inhibited by ≤1 μg of voriconazole per ml. Voriconazole was also active against
C. glabrata
(93% of the isolates were susceptible [MICs ≤ 1 μg/ml]) and
C. krusei
(100% of the isolates were susceptible). The agar-based Etest and disk diffusion methods performed well for the testing of both fluconazole and voriconazole compared to the broth microdilution MIC reference method. These observations establish the continued importance of
C. albicans
as a pathogen and the sustained activity of fluconazole and the broad spectrum of activity of voriconazole and will serve as the first-year benchmark for the ARTEMIS Program. Continued surveillance and refinement of broth- and agar-based test methods will help to identify susceptibility trends and improve the laboratory capability for antifungal susceptibility testing.
Molecular Analysis of the 18S rRNA Gene of
Cryptosporidium
Parasites from Patients with or without Human Immunodeficiency Virus Infections Living in Kenya, Malawi, Brazil, the United Kingdom, and Vietnam
Authors: Gatei Wangeci , Greensill Julie , Ashford Richard W. , Cuevas Luis E. , Parry Christopher M. , Cunliffe Nigel A. , Beeching Nicholas J. , Hart C. Anthony 
Published: April 2003

ABSTRACT

An 840-bp fragment of the 18S rRNA gene was used to identify
Cryptosporidium
spp. recovered from human immunodeficiency virus (HIV)-infected and -uninfected patients from Kenya, Malawi, Brazil, the United Kingdom, and Vietnam. Initial identification was by Ziehl-Neelsen acid-fast staining. Confirmation was by nested PCR, targeting the most polymorphic region of the 18S rRNA gene. Genotyping was by restriction endonuclease digestion of the PCR product followed by nucleotide sequencing. Among 63 isolates analyzed, four genotypes of
Cryptosporidium
were identified; 75% of the isolates were of the
C
.
parvum
human genotype, while the potentially zoonotic species were of the
C. parvum
bovine genotype (21.7%), the
C
.
meleagridis
genotype (1.6% [one isolate]), and the
C
.
muris
genotype (1.6% [one case]). HIV-infected individuals were more likely to have zoonotic genotypes than the HIV-uninfected individuals. Among the
C. parvum
group, strains clustered distinctly into either human or bovine genotypes regardless of the geographical origin, age, or HIV status of the patients. The intragenotypic variation observed in the
C. parvum
human genotype was extensive compared to that within the
C. parvum
bovine genotype group. The variation within genotypes was conserved in all geographical regions regardless of the patients' HIV status. The extensive diversity within genotypes at the 18S rRNA gene locus may limit its application to phylogenetic analyses.
DNA Fingerprinting ofSalmonella entericasubsp.entericaSerovar Typhimurium with Emphasis on Phage Type DT104 Based on Variable Number of Tandem Repeat Loci
Authors: Lindstedt Bjørn-Arne , Heir Even , Gjernes Elisabet , Kapperud Georg 
Published: April 2003

ABSTRACTSeventy-eight human and environmental strains ofSalmonella entericasubsp.entericaserovar Typhimurium, as well as 18 isolates of otherSalmonellaserovars and 6 isolates ofEscherichia coli, were subjected to a novel variable number of tandem repeats (VNTR)-based fingerprinting method that showed high discrimination and reproducibility for typing serovar Typhimurium isolates. The method is based on capillary separation of PCR products from fluorescence-labeled VNTR in the serovar Typhimurium genome. The serovar Typhimurium isolates displayed 54 VNTR patterns, and the VNTR assay correctly identified strains from a well-characterized outbreak. Among 37 serovar Typhimurium phage type DT104 isolates, 28 distinct VNTR patterns were found. This VNTR-based method is fast and suitable for complete automation. Our VNTR-based method was capable of high discrimination within the homogeneous serovar Typhimurium DT104 phage type and can be used to trace outbreaks and to monitor DT104 as well as other phage types. The VNTR assay was compared toXbaI pulsed-field gel electrophoresis, amplified fragment length polymorphism analysis, integron-cassette profiles and gene PCR ofintI1,qacEΔ1,sulI1, andfloR. The VNTR assay showed greatly improved resolution compared to all other tested methods in this study.
Development of a Duplex PCR Genotyping Assay for Distinguishing
Clostridium perfringens
Type A Isolates Carrying Chromosomal Enterotoxin (
cpe
) Genes from Those Carrying Plasmid-Borne Enterotoxin (
cpe
) Genes
Authors: Wen Qiyi , Miyamoto Kazuaki , McClane Bruce A. 
Published: April 2003

ABSTRACT

About 5% of
Clostridium perfringens
type A isolates carry the
cpe
gene encoding the
C. perfringens
enterotoxin. Those
cpe
-positive type A isolates are important causes of food-poisoning and non-food-borne cases of diarrheas in humans, as well as certain veterinary cases of diarrhea. Previous studies have determined that the enterotoxigenic type A isolates causing both non-food-borne human gastrointestinal disease and veterinary disease carry their
cpe
genes on plasmids, while the type A isolates causing human food poisoning carry a chromosomal
cpe
gene. The present study reports on the successful development of a duplex PCR assay that can rapidly genotype enterotoxigenic type A isolates (i.e., determine whether those
cpe
-positive isolates carry a chromosomal or a plasmid-borne
cpe
gene). The availability of this rapid
cpe
genotyping assay capable of handling large numbers of samples provides a powerful new investigative tool for diagnostic, epidemiologic, and basic research purposes.
Comparison of Ribotyping and Restriction Enzyme Analysis for Inter- and Intraspecies Discrimination of
Bordetella avium
and
Bordetella hinzii
Authors: Register Karen B. , Sacco Randy E. , Nordholm Gwen E. 
Published: April 2003

ABSTRACT

Bordetella avium
is an avian respiratory disease pathogen responsible for substantial economic losses to the turkey industry. The inability to distinguish isolates has hampered outbreak investigations and prevents a complete understanding of transmission mechanisms. Isolates of
Bordetella hinzii
, often referred to as
B. avium
-like or as
Alcaligenes faecalis
type II prior to 1995, have also been acquired from the respiratory tracts of diseased poultry but are not believed to be pathogenic for birds. Therefore, differentiating between
B. avium
and
B. hinzii
is of importance for veterinary diagnostic laboratories. It was recently reported that both
Pvu
II ribotyping and
Hin
fI/
Dde
I restriction endonuclease analysis (REA) show promise for distinguishing isolates of
B. avium
and
B. hinzii
. Here we compare the ability of these techniques to discern inter- and intraspecies differences. While both approaches distinguished numerous types within a species, only REA was sufficiently discriminatory for routine use as an epidemiologic tool. Both techniques clearly distinguish between
B. avium
and
B. hinzii
, although the results of ribotyping are more easily interpreted. Ribotyping and REA identified numerous, previously unrecognized
B. hinzii
strains from a collection of bordetella isolates, including one acquired from a rabbit. This is the first report of
B. hinzii
isolation from a nonhuman mammalian species. At least some of the newly recognized
B. hinzii
isolates have been previously reported to cause disease in poults, suggesting that the pathogenicity of this agent for poultry should be more rigorously examined.
Detection, Differentiation, and Quantitation of Pathogenic
Leishmania
Organisms by a Fluorescence Resonance Energy Transfer-Based Real-Time PCR Assay
Authors: Schulz Alexandra , Mellenthin Katja , Schönian Gabriele , Fleischer Bernhard , Drosten Christian 
Published: April 2003

ABSTRACT

Real-time technology eliminates many of the pitfalls of diagnostic PCR, but this method has not been applied to differentiation of
Leishmania
organisms so far. We have developed a real-time PCR that simultaneously detects, quantitates, and categorizes
Leishmania
organisms into three relevant groups causing distinct clinical pictures. The analytical sensitivity (detection rate of ≥95% at 94.1 parasites/ml of blood) was within a range that has been determined previously to facilitate the confirmation of visceral leishmaniasis from peripheral blood. Parasites were successfully detected in 12 different clinical samples (blood, bone marrow, skin, and liver). The
Leishmania donovani
complex, the
Leishmania brasiliensis
complex, and species other than these could be clearly discriminated by means of distinct melting temperatures obtained with fluorescence resonance energy transfer probes (melting points, 72.7, 67.1, and 65.0°C, respectively). All three groups could be quantified within equal ranges. As in other real-time PCRs, the variability in the quantification of DNA was small (coefficient of variation [CV], <2%). However, human samples containing low levels of parasites (100 parasites per ml of blood) showed higher variation (CV, 60.89%). Therefore, despite its superior analytical performance, care must be taken when real-time PCR is utilized for therapy monitoring.
International Typing Study of Toxin A-Negative, Toxin B-Positive
Clostridium difficile
Variants
Authors: Johnson Stuart , Sambol Susan P. , Brazier Jon S. , Delmée Michel , Avesani V. , Merrigan Michelle M. , Gerding Dale N. 
Published: April 2003

ABSTRACT

Clinically important strains of
Clostridium difficile
that do not produce toxin A but produce toxin B and are cytotoxic (A
−
/B
+
) have been reported from multiple countries. In order to compare the relatedness of these strains, we typed 23 A
−
/B
+
C. difficile
isolates from the United Kingdom (6 isolates), Belgium (11 isolates), and the United States (6 isolates) by three well-described typing methods. Restriction endonuclease analysis (REA), PCR ribotyping, and serogrouping differentiated 11, 4, and 3 different strain types, respectively. Twenty-one of the 23 A
−
/B
+
variants had a 1.8-kb truncation of the toxin A gene characteristic of toxinotype VIII strains; 20 of the 21 toxinotype VIII-like strains were PCR type 17. PCR type 17 isolates could be differentiated into two separate strain groups by serogrouping and by REA. REA further discriminated these isolates into eight subgroups (REA types). PCR type 17-serogroup F-REA group CF isolates were recovered from all three countries, and one specific REA type, CF4, was recovered from patients with
C. difficile
disease in the United Kingdom and the United States.
C. difficile
A
−
/B
+
variants of apparent clonal origin are widely distributed in Europe and North America.
Rapid Detection of Herpes Simplex Virus and Varicella-Zoster Virus Infections by Real-Time PCR
Authors: Weidmann Manfred , Meyer-König Ursula , Hufert Frank T. 
Published: April 2003

ABSTRACT
The herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2) and varicella-zoster virus (VZV) can cause life-threatening infections of the central nervous system and lead to severe infections in immunocompromised subjects and newborns. In these cases, rapid diagnosis is crucial. We developed three different real-time PCR assays based on TaqMan chemistry for the LightCycler instrument to detect HSV-1, HSV-2, and VZV. When the TaqMan assays were compared to our in-house nested PCR assays, the test systems had equal sensitivities of ≤10 plasmid copies per assay. When clinical samples were investigated by TaqMan PCR to detect HSV-1, HSV-2, and VZV DNA, 95, 100, and 96% of the samples determined to be positive by nested PCR, respectively, were positive by the real-time PCR assays. The specificities of all PCR assays were almost 100%. Furthermore, the TaqMan PCR assays could be performed within 2.5 h, whereas nested PCR results were available after 9 h. In addition to offering more rapid results, the TaqMan PCR assays appear to be less expensive than nested PCR assays due to less hands-on time. In summary, TaqMan PCR is an excellent alternative to conventional nested PCR assays for the rapid detection of HSV-1, HSV-2, and VZV in clinical samples.
Evolution of a Vancomycin-Intermediate
Staphylococcus aureus
Strain In Vivo: Multiple Changes in the Antibiotic Resistance Phenotypes of a Single Lineage of Methicillin-Resistant
S
.
aureus
under the Impact of Antibiotics Administered for Chemotherapy
Authors: Sieradzki K. , Leski T. , Dick J. , Borio L. , Tomasz A. 
Published: April 2003

ABSTRACT

A number of methicillin-resistant
Staphylococcus aureus
(MRSA) isolates were recovered over a period of several weeks from blood samples and from the heart valve of a patient who underwent extensive vancomycin chemotherapy for persistent
S
.
aureus
bacteremia. Consecutive isolates showed gradually decreasing growth rates during in vitro cultivation and increasing vancomycin MICs, from an MIC of 1 μg/ml for the initial isolate to an MIC of 8 μg/ml for the final MRSA isolates, which also became tolerant to vancomycin. Major changes were observed in the oxacillin resistance phenotype of several of the isolates—apparently related to in vivo exposure to imipenem, which was also used during a period of chemotherapy. Both the gradually increasing vancomycin MICs and the changes in oxacillin resistance could be reproduced by appropriate exposure of the initial MRSA isolate to antibiotics in vitro. All isolates had the same pulsed-field gel electrophoresis pattern,
spaA
type, and multilocus sequence type (MLST), which was identified as a single-locus variant of ST5, the MLST characteristic of previously characterized MRSA isolates with reduced susceptibility to vancomycin in the United States and Japan.
Blood Agar and
Mycobacterium tuberculosis
: the End of a Dogma
Authors: Drancourt M. , Carrieri P. , Gévaudan M.-J. , Raoult D. 
Published: April 2003

ABSTRACT

Incidental blood agar-based recovery of
Mycobacterium tuberculosis
led us to further investigate this routine medium for primary isolation and culture of
M. tuberculosis
. Fifteen respiratory tract and eight lymph node Ziehl-Neelsen-positive specimens were inoculated in parallel into tubes containing egg-based medium and 5% sheep blood agar. Colonies appeared sooner on this medium than on the egg-based medium, but this difference was not significant (
P
= 0.11, analysis of variance [ANOVA] test). Further experiments compared the growth of 38 respiratory and lymph node
M. tuberculosis
isolates when subcultured on the two media. After 6 days of incubation, 21 of 38 isolates had grown on blood agar, and the mean number of colonies was significantly greater on blood agar than on the egg-based medium (
P
< 0 0.001, ANOVA test). These results demonstrate that
M. tuberculosis
grows easily on blood agar within 1to 2 weeks, indicating that this basic medium is suitable for laboratory diagnosis of tuberculosis in addition to other media. Laboratories that routinely use prolonged incubations of blood plates, for example, for the recovery of
Bartonella
species, should consider the potential safety implications of encountering this highly infectious pathogen.
Enterobacterial Repetitive Intergenic Consensus 1R PCR Assay for Detection of
Raoultella
sp. Isolates among Strains Identified as
Klebsiella oxytoca
in the Clinical Laboratory
Authors: Granier Sophie A. , Leflon-Guibout Véronique , Goldstein Fred W. , Nicolas-Chanoine Marie-Hélène 
Published: April 2003

ABSTRACT

The enterobacterial repetitive intergenic consensus 1R PCR method, which provided recognizable profiles for reference strains of the three species of
Raoultella
and the two genetic groups of
Klebsiella oxytoca
, was applied to 19 clinical isolates identified as
K. oxytoca
. By this method, as confirmed by species-specific gene sequencing, two
Raoultella ornithinolytica
and two unclassifiable
K. oxytoca
isolates were identified.