bluuebunny/crossref_metadata_2025_split huggingface dataset:
Detection of Human Herpesvirus 7 DNA by Loop-Mediated Isothermal Amplification
Authors: Yoshikawa Tetsushi , Ihira Masaru , Akimoto Shiho , Usui Chie , Miyake Fumi , Suga Sadao , Enomoto Yoshihiko , Suzuki Ryota , Nishiyama Yukihiro , Asano Yoshizo 
Published: March 2004

ABSTRACT
The reliability of loop-mediated isothermal amplification (LAMP), initially developed for the detection of human herpesvirus 7 (HHV-7), was evaluated in this study. Although a LAMP product was detected in HHV-7 DNA, neither HHV-6 nor human cytomegalovirus DNA produced a product. When agarose gel electrophoresis was used for the detection of LAMP products, the sensitivity of a 30-min HHV-7 LAMP reaction reached 250 copies/tube. The use of turbidity for the detection of the LAMP products gave a sensitivity of 500 and 250 copies/tube for 30- and 60-min reactions, respectively. Following these initial validation studies, clinical samples collected from two patients with primary HHV-7 infections were examined by HHV-7 LAMP. By use of agarose gel electrophoresis, HHV-7 LAMP products could be detected in acute-phase plasma samples but no LAMP product was detectable in convalescent-phase plasma samples from either patient. Since a turbidity assay is less sensitive than agarose gel electrophoresis, no HHV-7 LAMP product could be detected in plasma samples after a 30-min LAMP reaction. After a 60-min LAMP reaction, HHV-7 LAMP product could be detected in acute-phase plasma samples.
Detection of Ehrlichial DNA in
Haemaphysalis
Ticks Recovered from Dogs in Japan That Is Closely Related to a Novel
Ehrlichia
sp. Found in Cattle Ticks from Tibet, Thailand, and Africa
Authors: Inokuma Hisashi , Beppu Takeshi , Okuda Masaru , Shimada Yojiro , Sakata Yoshimi 
Published: March 2004

ABSTRACT

Tick DNA samples from dogs in Japan were examined for
Ehrlichia
infection by 16S rRNA gene-based PCR and sequencing. Three positive samples were detected from
Haemaphysalis
ticks, and higher levels of similarity (98.46 to 99.06%) were found to recently detected
Ehrlichia
spp. from cattle ticks in Tibet, Thailand, and Africa.
Cryptococcus neoformans
Shows a Remarkable Genotypic Diversity in Brazil
Authors: Barreto de Oliveira M. T. , Boekhout T. , Theelen B. , Hagen F. , Baroni F. A. , Lazera M. S. , Lengeler K. B. , Heitman J. , Rivera I. N. G. , Paula C. R. 
Published: March 2004

ABSTRACT

The genotypic diversity of Brazilian
Cryptococcus neoformans
strains was analyzed. The majority of the samples were αA (65%), followed by αB (17.5%), αD (9%), αAaD hybrids (5%), and αC (3.5%). A considerable genotypic diversity occurred within
C. neoformans
var.
grubii
, and a new amplified fragment length polymorphism genotype, 1B, was recognized.
Legionella pneumophila
Serogroup 4 Isolated from Joint Tissue
Authors: Linscott Andrea J. , Poulter Melinda D. , Ward Kevin , Bruckner David A. 
Published: March 2004

ABSTRACT

We report the isolation of
Legionella pneumophila
serogroup 4 from synovial tissue obtained from an 80-year-old female with chronic swelling of her right metacarpophalangeal joint. Synovial tissue infections caused by
L. pneumophila
are rare. Interestingly, this isolate was recovered from chocolate agar after 5 days of incubation.
Molecular Epidemiology of Sequential Outbreaks of
Acinetobacter baumannii
in an Intensive Care Unit Shows the Emergence of Carbapenem Resistance
Authors: Zarrilli Raffaele , Crispino Margherita , Bagattini Maria , Barretta Elena , Di Popolo Anna , Triassi Maria , Villari Paolo 
Published: March 2004

ABSTRACT

The molecular epidemiology of multidrug-resistant
Acinetobacter baumannii
was investigated in the medical-surgical intensive care unit (ICU) of a university hospital in Italy during two window periods in which two sequential
A. baumannii
epidemics occurred. Genotype analysis by pulsed-field gel electrophoresis (PFGE) of
A. baumannii
isolates from 131 patients identified nine distinct PFGE patterns. Of these, PFGE clones B and I predominated and occurred sequentially during the two epidemics.
A. baumannii
epidemic clones showed a multidrug-resistant antibiotype, being clone B resistant to all antimicrobials tested except the carbapenems and clone I resistant to all antimicrobials except ampicillin-sulbactam and gentamicin. Type 1 integrons of 2.5 and 2.2 kb were amplified from the chromosomal DNA of epidemic PFGE clones B and I, respectively, but not from the chromosomal DNA of the nonepidemic clones. Nucleotide analysis of clone B integron identified four gene cassettes:
aacC1
, which confers resistance to gentamicin; two open reading frames (ORFs) coding for unknown products; and
aadA1a
, which confers resistance to spectinomycin and streptomycin. The integron of clone I contained three gene cassettes:
aacA4
, which confers resistance to amikacin, netilmicin, and tobramycin; an unknown ORF; and
bla
OXA-20
, which codes for a class D β-lactamase that confers resistance to amoxicillin, ticarcillin, oxacillin, and cloxacillin. Also, the
bla
IMP
allele was amplified from chromosomal DNA of
A. baumannii
strains of PFGE type I. Class 1 integrons carrying antimicrobial resistance genes and
bla
IMP
allele in
A. baumannii
epidemic strains correlated with the high use rates of broad-spectrum cephalosporins, carbapenems, and aminoglycosides in the ICU during the study period.
Analysis of the Serotype and Genotype Correlation of VP1 and the 5′ Noncoding Region in an Epidemiological Survey of the Human Enterovirus B Species
Authors: Thoelen Inge , Moës Elien , Lemey Philippe , Mostmans Sara , Wollants Elke , Lindberg A. Michael , Vandamme Anne-Mieke , Van Ranst Marc 
Published: March 2004

ABSTRACT
The sequence identity of the enterovirus VP1 gene has been shown to correlate with the serotype concept. Enterovirus molecular typing methods are therefore often based on sequencing of the VP1 genomic region and monophyletic clustering of VP1 sequences of a homologous serotype. For epidemiological surveillance, 342 enterovirus samples obtained from patients with aseptic meningitis in Belgium from 1999 to 2002 were first diagnosed as being enterovirus positive by amplification of the 5′ noncoding region (5′NCR) by reverse transcription (RT)-PCR. Subsequently, samples were molecularly typed by RT-nested PCR amplification and sequencing of a portion of the VP1 gene. Phylogenetic analyses were performed to investigate enteroviral evolution and to examine the serotype and genotype correlation of the two genomic regions. Our typing results demonstrated echovirus 30, echovirus 13, echovirus 18, and echovirus 6 to be the most predominant types. Echoviruses 13 and 18 were considered to be emerging human serotypes since 2000 and 2001, respectively, as they had been rarely reported before. Several serotypes existed as multiple genotypes (subtypes) from 1999 to 2002, but genomic differences mainly resided at synonymous sites; these results strongly suggest that the subtypes exhibit similar antigenic properties. Phylogenetic analyses confirmed that VP1 is an adequate region for molecular typing. Serotype-specific clusters are not observed commonly in phylogenetic trees based on the 5′NCR, and the phylogenetic signal in the 5′NCR was found to be particularly low. However, some substructure in the 5′NCR tree made a tentative prediction of the enterovirus type possible and was therefore helpful in PCR strategies for VP1 (e.g., primer choice), provided some background knowledge on the local spectrum of enteroviruses already exists.
Preclinical Evaluation of Two Real-Time, Reverse Transcription-PCR Assays for Detection of the Severe Acute Respiratory Syndrome Coronavirus
Authors: Bressler Adam M. , Nolte Frederick S. 
Published: March 2004

ABSTRACT
We verified the analytical performance characteristics of a previously described real-time reverse transcription-PCR (RT-PCR) assay targeting the open reading frame (ORF) 1b region of the severe acute respiratory syndrome coronavirus (SARS-CoV) with RNA transcripts. We then compared it to a novel nucleocapsid gene real-time RT-PCR assay with genomic RNA. The assays differed only in the primer and probe sequences and final concentrations. A commercially available armored RNA (Ambion, Austin, Tex.) was evaluated as positive control for the ORF 1b assay. The analytical sensitivity, reproducibility, amplification efficiency, and dynamic range of the assays were similar. Both were specific for SARS-CoV as determined by testing against human CoV 229E and OC43, specimens from patients without SARS, and by BLAST searches of GenBank for primer and probe sequence homology. The armored RNA was found to be a suitable positive control for the ORF 1b assay that could be reliably recovered and amplified from a variety of clinical specimens.
Molecular Epidemiology of Coagulase-Negative Staphylococci Causing Sepsis in a Neonatal Intensive Care Unit over an 11-Year Period
Authors: Krediet Tannette G. , Mascini Ellen M. , van Rooij Ellen , Vlooswijk Judith , Paauw Armand , Gerards Leo J. , Fleer André 
Published: March 2004

ABSTRACT

Coagulase-negative staphylococci (CoNS) are the major causative microorganisms in neonatal nosocomial sepsis. Previous studies have shown that CoNS sepsis in the neonatal intensive care unit (NICU) is caused by predominant molecular types that are widely distributed among both neonates and staff. Some of these molecular types may persist in the NICU for years. The purpose of the present study was to determine the dynamic behavior of CoNS strains causing sepsis over a prolonged period of time by determining the molecular types of all blood isolates from septicemic infants over a period of 11 years (1991 to 2001). The results show that neonatal CoNS sepsis is increasingly caused by a few predominant molecular clusters. The most striking finding was that in recent years one molecular cluster emerged as the predominant cause of neonatal CoNS sepsis, responsible for no less than 31% (20 of 65) of blood isolates in 2001. Antibiotic resistance, particularly beta-lactam resistance, is probably an important selective force considering the high
mecA
gene carriage of CoNS blood isolates (70 to 92%). We conclude that neonatal CoNS sepsis is increasingly caused by a limited number of predominant molecular CoNS types and that antibiotic resistance is probably a major selective force.
Comparison of Conventional Bacteriology with Nucleic Acid Amplification (Amplified Mycobacterium Direct Test) for Diagnosis of Tuberculous Meningitis before and after Inception of Antituberculosis Chemotherapy
Authors: Thwaites Guy E. , Caws Maxine , Chau Tran Thi Hong , Dung Nguyen Thi , Campbell James I. , Phu Nguyen Hoan , Hien Tran Tinh , White Nicholas J. , Farrar Jeremy J. 
Published: March 2004

ABSTRACT

The role of nucleic acid amplification techniques in the rapid diagnosis of tuberculous meningitis remains uncertain. We compared the performance of Ziehl-Neelsen (ZN) staining, the Gen-Probe amplified
Mycobacterium tuberculosis
direct test (MTD), and culture with 341 cerebrospinal fluid specimens from 152 adults (73 with and 79 without tuberculous meningitis) before and after inception of antituberculosis chemotherapy. The sensitivity, specificity, and positive and negative predictive values of ZN staining before treatment were 34/66 (52%), 79/79 (100%), 34/34 (100%), and 79/111 (71%), compared with 25/66 (38%), 78/79 (99%), 25/26 (96%), and 79/120 (66%) for MTD. The sensitivity of combined ZN staining and MTD (either positive) was 45/66 (68%). The sensitivity of staining and culture fell more rapidly than that of MTD after the start of treatment: after 5 to 15 days of treatment, MTD was more sensitive than ZN staining (12/43 [28%] versus 2/43 [2%];
P
= 0.013). Slower bacterial clearance was observed if
M. tuberculosis
was resistant to isoniazid and/or streptomycin: resistant organisms were more likely to be cultured from cerebrospinal fluid after 2 to 5 days of treatment than fully sensitive organisms (
P
< 0.001). The sensitivities of ZN staining, MTD, and the two tests combined were improved by repeated sampling to 38/59 (64%), 35/59 (59%), and 49/59 (83%), respectively. In conclusion, ZN staining of the cerebrospinal fluid is at least as good as MTD for the rapid diagnosis of tuberculosis and is much faster and less expensive. However, the combination of these methods on serial samples detects more cases. Alternative tests are still urgently required.
Studies on the genus Paraphanolaimus Micoletzky, 1923 (Nematoda: Aphanolaimidae) with description of P. paraguayensis sp. n.
Authors: Holovachov Oleksandr , Sturhan Dieter 
Published: March 2004

Abstract
Two known and one new species of Paraphanolaimus are described, viz. P.
behningi from Germany, P. anisitsi from Paraguay and Germany and P.
paraguayensis sp. n. from Paraguay. Paraphanolaimus paraguayensis sp. n. is
characterised by 0.84-0.94 mm long body, presence of 37-44 lateral epidermal
glands, strongly sclerotised and broad cylindrical stoma, swollen tail tip;
female reproductive system with developed spermathecae, straight vagina;
male without neck setae, 16-19 tubular supplements, two precloacal and nine
to ten caudal setae, spicules 32-33 μm long, arcuate, with oval manubrium
and fusiform shaft, platelike gubernaculum. Paraphanolaimus indicus and
P.micoletzkyi are considered as species inquirendae. An emended diagnosis
and a revised classification of Paraphanolaimus are proposed and a key to
the species of the genus is provided. Aphanonchus bayensis (Keppner, 1988)
comb. n. is proposed for Paraphanolaimus bayensis Keppner, 1988.
Purification of two substances from bulbs of onion (Allium cepa L.) with nematicidal activity against Meloidogyne exigua Goeldi
Authors: Amaral Daniel Rufino , Rocha Oliveira Fernanda Emanuele da , Oliveira Denilson Ferreira , Campos Vicente Paulo 
Published: March 2004

Abstract
To contribute to the development of a new method for Meloidogyne exigua
Goeldi control, a nematode widely disseminated through Brazilian coffee
fields, this work aimed to purify the nematicidal substances produced by
onion (Allium cepa L.). Thus, the methanolic extract of onion bulbs was
concentrated under reduced pressure and submitted to a purification process
guided by in vitro assays with second-stage juveniles (J2) of M. exigua.
After several steps, two nematicidal substances that seemed to be
interconverting when in solution were isolated. When exposed to 500 ppm
aqueous solutions of substances 1 and 2, 55 and 38% of the J2, respectively,
were dead after 24 h. Very preliminary nuclear magnetic resonance
spectroscopy studies suggested that these substances correspond to a pair of
carbohydrates.
Pre-parasitic juveniles of Strelkovimermis spiculatus Poinar & Camino, 1986 (Nematoda: Mermithidae) predated upon by freshwater copepods (Crustacea: Copepoda)
Authors: Achinelly María , Micieli María , García Juan 
Published: March 2004

Abstract
Five copepod species, Acantocyclops robustus, Macrocyclops albidus,
Mesocyclops annulatus, Thermocyclops sp. (all Cyclopoidea) and
Argyrodiaptomus bergi (Calanoidea), were evaluated under laboratory
conditions as potential predators of preparasites of the mermithid nematode
Strelkovimermis spiculatus. Adults of all five copepod species consumed
70-100% of the preparasites within 24 h. Copepodids and adults of M.
annulatus predated upon 93 and 100% of the nematode pre-parasites,
respectively. An average of 197 pre-parasites was daily predated upon by an
adult M. annulatus. Copepod density affected prevalence and intensity of S.
spiculatus on mosquito larvae. Prevalence and intensity of S. spiculatus on
Aedes aegypti larvae was reduced from 98% and 2.8 nematodes per larva in
control containers free of copepods to 61.4% and 1.4 nematodes per larva in
containers with 50 M. annulatus females per l.
Distribution and pathogenicity of bacteria species carried by Bursaphelenchus xylophilus in China
Authors: Zhao Bo Guang , Wang Hu Li , Han Su Fen , Han Zheng Ming 
Published: March 2004

Abstract
Bacteria carried by the pine wood nematode (PWN), Bursaphelenchus xylophilus,
from both healthy and diseased Pinus thunbergii and P. massoniana were
studied in five main pine wilt disease epidemic provinces in P. R. China. No
bacteria were found in healthy pines but were found on PWN from all samples
collected from diseased trees. Twenty-four bacteria strains were isolated
from the nematodes and were identified by a combination of classical and
automatic testing bacteriology (ATB) expression methods. Bioassay showed
that 17 of the 24 identified strains produced phytotoxins. Eleven of these
17 phytotoxin producers belonged to the genus Pseudomonas. Glasshouse and
field inoculation tests using sterile techniques showed that both PWN and
the toxin-producing bacteria carried were necessary to induce disease. We
hypothesise that pine wilt disease is a complex, induced by both PWN and the
bacteria it carries.
A Novel Dysfunctional Growth Hormone Variant (Ile179Met) Exhibits a Decreased Ability to Activate the Extracellular Signal-Regulated Kinase Pathway
Authors: Lewis Mark D. , Horan Martin , Millar David S. , Newsway Vicky , Easter Tammy E. , Fryklund Linda , Gregory John W. , Norin Martin , Del Valle Cristóbal-Jorge , López-Siguero Juan Pedro , Cañete Ramón , López-Canti Luis Fernando , Díaz-Torrado Nieves , Espino Rafael , Ulied Angels , Scanlon Maurice F. , Procter Annie M. , Cooper David N. 
Published: March 2004

Abstract
The pituitary-expressed GH1 gene was screened for mutation in a group of 74 children with familial short stature. Two novel mutations were identified: an Ile179Met substitution and a −360A→G promoter variant. The Ile179Met variant was shown to exhibit a similar degree of resistance to proteolysis as wild-type GH, indicating that the introduction of Met does not cause significant misfolding. Secretion of Ile179Met GH from rat pituitary cells was also similar to that of wild type. Although receptor binding studies failed to show any difference in binding characteristics, molecular modeling studies suggested that the Ile179Met substitution might nevertheless perturb interactions between GH and the GH receptor loop containing the hotspot residue Trp169, thereby affecting signal transduction. The ability of the Ile179Met variant to activate a signal transducer and activator of transcription (STAT) 5-responsive luciferase reporter gene and induce phosphorylation of STAT 5 and ERK was therefore studied. In contrast to its ability to activate STAT 5 normally, activation of ERK by the Ile179Met variant was reduced to half that observed with wild type. Although differential effects on the activation of distinct signaling pathways by a mutant receptor agonist are unprecedented, these findings also suggest that the ERK pathway could play a role in mediating the action of GH.
Failure of Mathematical Indices to Accurately Assess Insulin Resistance in Lean, Overweight, or Obese Women with Polycystic Ovary Syndrome
Authors: Diamanti-Kandarakis Evanthia , Kouli Chryssa , Alexandraki Krystallenia , Spina Giovanna 
Published: March 2004

Abstract
Insulin resistance is a common metabolic feature of polycystic ovary syndrome (PCOS). In this study, we examined the validity of the mathematical indices [the quantitative insulin sensitivity check index (QUICKI) and the homeostasis model of assessment (HOMA)] that calculate insulin sensitivity and their correlation to glucose utilization with the insulin infusion rate in 40 mU/m2·min by the euglycemic clamp (M) in women with PCOS.
We studied 59 women with PCOS (20 lean, 16 overweight, and 23 obese subjects). Euglycemic clamp testing was performed, and QUICKI, HOMA, total testosterone, fasting insulin, fasting glucose, and glucose-to-insulin ratio were estimated.
No difference was found in testosterone and glucose levels among the three groups. Lean or overweight women compared with obese women differed in insulin levels, glucose-to-insulin ratio, QUICKI, and HOMA (P &lt; 0.01). No statistical difference was found between lean and overweight women in the above parameters. M differed when lean women were compared with overweight (P &lt; 0.002) or obese women (P &lt; 0.0001); however, no statistical difference was observed between overweight and obese women. No significant correlation was found between M and QUICKI or HOMA.
We conclude that mathematical indices should be applied with caution in different insulin-resistant populations and should not be considered a priori equivalent to the euglycemic clamp technique.
Promoter-Specific Repression of Hepatocyte Nuclear Factor (HNF)-1β and HNF-1α Transcriptional Activity by an HNF-1β Missense Mutant Associated with Type 5 Maturity-Onset Diabetes of the Young with Hepatic and Biliary Manifestations
Authors: Kitanaka Sachiko , Miki Yuko , Hayashi Yasuhide , Igarashi Takashi 
Published: March 2004

AbstractMutations in the hepatocyte nuclear factor (HNF)-1β lead to type 5 maturity-onset diabetes of the young (MODY5). HNF-1β forms a homodimer or a heterodimer with HNF-1α and regulates various target genes. HNF-1β mutations are rare, and no functional analysis has been performed in conjunction with HNF-1α. HNF-1β is expressed in the liver and biliary system and controls liver-specific and bile acid-related genes. Moreover, liver-specific Hnf-1β knockout mice present with severe jaundice. However, no patients with HNF-1β mutations have biliary manifestations. In this report, we found a novel missense mutation in the HNF-1β gene in a patient with neonatal cholestasis and liver dysfunction together with the common features of MODY5. Functional analysis revealed that the mutant HNF-1β had diminished transcriptional activity by loss of the DNA binding activity. The mutant had a promoter-specific dominant-negative transcriptional effect on wild-type HNF-1β and inhibited its DNA binding. Moreover, the mutant had a promoter- and cell-specific transcriptional repressive effect on HNF-1α and a promoter-specific inhibitory effect on HNF-1α DNA binding. From these results, we considered that the different phenotype of patients with HNF-1β mutations might be caused by the different HNF-1β activity in conjunction with the different repression of HNF-1α activity in selected promoters and tissues.
Increased Diagnosis of Primary Aldosteronism, Including Surgically Correctable Forms, in Centers from Five Continents
Authors: Mulatero Paolo , Stowasser Michael , Loh Keh-Chuan , Fardella Carlos E. , Gordon Richard D. , Mosso Lorena , Gomez-Sanchez Celso E. , Veglio Franco , Young William F. 
Published: March 2004

Abstract
Primary aldosteronism (PA) is a common form of endocrine hypertension previously believed to account for less than 1% of hypertensive patients. Hypokalemia was considered a prerequisite for pursuing diagnostic tests for PA. Recent studies applying the plasma aldosterone/plasma renin activity ratio (ARR) as a screening test have reported a higher prevalence. This study is a retrospective evaluation of the diagnosis of PA from clinical centers in five continents before and after the widespread use of the ARR as a screening test. The application of this strategy to a greater number of hypertensives led to a 5- to 15-fold increase in the identification of patients affected by PA. Only a small proportion of patients (between 9 and 37%) were hypokalemic. The annual detection rate of aldosterone-producing adenoma (APA) increased in all centers (by 1.3–6.3 times) after the wide application of ARR. Aldosterone-producing adenomas constituted a much higher proportion of patients with PA in the four centers that employed adrenal venous sampling (28–50%) than in the center that did not (9%). In conclusion, the wide use of the ARR as a screening test in hypertensive patients led to a marked increase in the detection rate of PA.