bluuebunny/crossref_metadata_2025 huggingface dataset:
Genomic Approach to Identification of
Mycobacterium bovis
Diagnostic Antigens in Cattle
Authors: Aagaard Claus , Govaerts Marc , Meng Okkels Limei , Andersen Peter , Pollock John M.
Published: August 2003

Differential delayed-type hypersensitivity skin testing with tuberculin purified protein derivatives from
Mycobacterium bovis
and
M. avium
is the standard for diagnosing bovine tuberculosis. However, improved tests based on defined, specific antigens are urgently needed. In the present study, a combination of bioinformatics, molecular biology, and bovine models of infection were used to screen mycobacterial proteins for their potential as diagnostic reagents which could be used in a whole-blood assay for diagnosis of tuberculosis. Initial screening of 28 proteins selected in silico and expressed as recombinants in
Escherichia coli
indicated that CFP-10, ESAT-6, TB27.4, TB16.2, TB15.8, and TB10.4 induced strong gamma interferon responses in experimentally infected cattle. A more thorough investigation over time in two groups of animals infected with a high (10
6
CFU) and a low (10
4
CFU) dose of
M. bovis
revealed that, for both groups, the strength of the in vitro response to individual antigens varied greatly over time. However, combining the results for ESAT-6, CFP-10, and TB27.4, possibly supplemented with TB10.4, gave sensitivities at different infection stages close to those obtained with
M. bovis
purified protein derivative. Importantly, while responsiveness to ESAT-6 and CFP-10 correlated strongly for individual samples, the same was not the case for ESAT-6 and TB27.4 responsiveness. The results suggest that combinations of specific antigens such as these have great potential in development of optimized diagnostic systems for bovine tuberculosis.
Geographic Diversity among Genotypes of
Entamoeba histolytica
Field Isolates
Authors: Haghighi Ali , Kobayashi Seiki , Takeuchi Tsutomu , Thammapalerd Nitaya , Nozaki Tomoyoshi
Published: August 2003

It has been known that only 5 to 10% of those infected with
Entamoeba histolytica
develop symptomatic disease. However, the parasite and the host factors that determine the onset of disease remain undetermined. Molecular typing by using polymorphic genetic loci has been proven to aid in the close examination of the population structure of
E. histolytica
field isolates in nature. In the present study, we analyzed the genetic polymorphisms of two noncoding loci (locus 1-2 and locus 5-6) and two protein-coding loci (chitinase and serine-rich
E. histolytica
protein [SREHP]) among 79 isolates obtained from different geographic regions, mainly Japan, Thailand, and Bangladesh. When the genotypes of the four loci were combined for all isolates that we have analyzed so far (overlapping isolates from mass infection events were excluded), a total of 53 different genotypes were observed among 63 isolates. The most remarkable and extensive variations among the four loci was found in the SREHP locus; i.e., 34 different genotypes were observed among 52 isolates. These results demonstrate that
E. histolytica
has an extremely complex genetic structure independent of geographic location. Our results also show that, despite the proposed transmission of other sexually transmitted diseases, including human immunodeficiency virus infection, from Thailand to Japan, the spectra of the genotypes of the
E. histolytica
isolates from these two countries are distinct, suggesting that the major
E. histolytica
strains prevalent in Japan at present were likely introduced from countries other than Thailand. Although the genetic polymorphism of the SREHP locus was previously suggested to be closely associated with the clinical presentation, e.g., colitis or dysentery and liver abscess, no association between the clinical presentation and the SREHP genotype at either the nucleotide or the predicted amino acid level was demonstrated.
Presence of Activatable Shiga Toxin Genotype (
stx
2d
) in Shiga Toxigenic
Escherichia coli
from Livestock Sources
Authors: Gobius Kari S. , Higgs Glen M. , Desmarchelier Patricia M.
Published: August 2003

Stx2d is a recently described Shiga toxin whose cytotoxicity is activated 10- to 1,000-fold by the elastase present in mouse or human intestinal mucus. We examined Shiga toxigenic
Escherichia coli
(STEC) strains isolated from food and livestock sources for the presence of activatable
stx
2d
. The
stx
2
operons of STEC were first analyzed by PCR-restriction fragment length polymorphism (RFLP) analysis and categorized as
stx
2
,
stx
2c vha
,
stx
2c vhb
, or
stx
2d EH250
. Subsequently, the
stx
2c vha
and
stx
2c vhb
operons were screened for the absence of a
Pst
I site in the
stx
2A
subunit gene, a restriction site polymorphism which is a predictive indicator for the
stx
2d
(activatable) genotype. Twelve STEC isolates carrying putative
stx
2d
operons were identified, and nucleotide sequencing was used to confirm the identification of these operons as
stx
2d
. The complete nucleotide sequences of seven representative
stx
2d
operons were determined. Shiga toxin expression in
stx
2d
isolates was confirmed by immunoblotting.
stx
2d
isolates were induced for the production of bacteriophages carrying
stx
. Two isolates were able to produce bacteriophages φ1662a and φ1720a carrying the
stx
2d
operons. RFLP analysis of bacteriophage genomic DNA revealed that φ1662a and φ1720a were highly related to each other; however, the DNA sequences of these two
stx
2d
operons were distinct. The STEC strains carrying these operons were isolated from retail ground beef. Surveillance for STEC strains expressing activatable Stx2d Shiga toxin among clinical cases may indicate the significance of this toxin subtype to human health.
Evolution of Sporadic Isolates of Methicillin-Resistant
Staphylococcus aureus
(MRSA) in Hospitals and Their Similarities to Isolates of Community-Acquired MRSA
Authors: Aires de Sousa M. , de Lencastre H.
Published: August 2003

Forty-one methicillin-resistant
Staphylococcus aureus
(MRSA) hospital isolates that clearly differed from the six major pandemic clones of MRSA in pulsed-field gel electrophoresis type,
mecA
and Tn
554
polymorphism, and epidemic behavior were selected from an international strain collection for more detailed characterization.
SpaA
typing, multilocus sequence typing, and SCC
mec
(staphylococcal cassette chromosome
mec
) typing demonstrated extensive diversity among these sporadic isolates both in genetic background and also in the structure of the associated SCC
mec
elements. Nevertheless, the isolates could be grouped into restricted clonal complexes by using the BURST (i.e., based upon related sequence types) program algorithm, which predicted that most sporadic MRSA isolates evolved from pandemic MRSA clones. Several of the sporadic MRSA resembled community-acquired MRSA isolates in properties that included a relatively limited multiresistance pattern, faster growth rates, diversity of genetic backgrounds, and a frequent association with SCC
mec
type IV.
Differences in Gene Content among
Salmonella enterica
Serovar Typhi Isolates
Authors: Boyd E. F. , Porwollik S. , Blackmer F. , McClelland M.
Published: August 2003

We used a nonredundant microarray of the
Salmonella enterica
serovar Typhimurium LT2 and Typhi CT18 genomes to assess the genomic content of a diverse set of isolates of serovar Typhi. Comparative genomic hybridization revealed 13 regions of absent or divergent gene content in the eight Typhi strains examined compared to Typhi CT18. In particular, two Typhi CT18 prophage regions, STY1048 to STY1077 and STY2038 to STY2077, as well as a five-gene islet (STY3188 to STY3193) were absent or divergent in all other Typhi strains examined. Seven Typhi strains lacked most or all of the IS
1
elements present in strain CT18, and three Typhi strains lacked a P4-like phage (STY4821 to STY4834). One strain was devoid of a 149-gene region (STY4521 to STY4680), which encodes numerous phage genes and the Vi antigen biosynthesis and export gene cluster, a type IV pilus, and numerous phage genes. In Typhi strain 26T25, an amplification of an entire inter-ribosomal region encompassing 31 genes has occurred. Furthermore, a 257-gene region (STY1360 to STY1639) showed an aberrant replication pattern in three Typhi isolates. Overall, these differences in gene content indicate that even within a highly clonal bacterial population the genomic reservoir is unstable.
Evaluation of the Rapid Immunoassay Determine HIV 1/2 for Detection of Antibodies to Human Immunodeficiency Virus Types 1 and 2
Authors: van den Berk G. E. L. , Frissen P. H. J. , Regez R. M. , Rietra P. J. G. M.
Published: August 2003

We evaluated the reliability of a rapid human immunodeficiency virus type 1 test for quick clinical decision making, such as in needle-stick accidents. The test was evaluated with 1,160 patients. It proved to be a simple and useful test with 99.6% specificity and 99.4% sensitivity. One patient with late-stage AIDS had a false-negative result.
Evaluation of Susceptibility of Human Herpesvirus 8 to Antiviral Drugs by Quantitative Real-Time PCR
Authors: Sergerie Yan , Boivin Guy
Published: August 2003

A new in vitro system based on real-time PCR was developed for evaluation of human herpesvirus 8 susceptibility to antiviral agents. Cidofovir had the greatest inhibitory activity against HHV-8 (50% inhibitory concentration [IC
50
], 0.43 μM) followed by ganciclovir (2.61 μM), adefovir (18.00 μM), acyclovir (31.00 μM), and foscarnet (34.15 μM). The potential therapeutic efficacy for HHV-8 (i.e., peak serum drug level/IC
50
) is highest for cidofovir (167) and foscarnet (22).
Analysis of the Ability of Spirochete Species Associated with Relapsing Fever, Avian Borreliosis, and Epizootic Bovine Abortion To Bind Factor H and Cleave C3b
Authors: McDowell John V. , Tran Emily , Hamilton Duncan , Wolfgang Jill , Miller Kelley , Marconi Richard T.
Published: August 2003

Some
Borrelia
species associated with Lyme disease bind the complement-regulatory protein factor H (fH), a process that may aid in immune evasion. In this report we demonstrate that some
Borrelia
species associated with relapsing fever bind fH, but not those associated with avian borreliosis and epizootic bovine abortion. Cell-bound fH was also found to mediate cleavage of exogenously supplied human C3b, demonstrating the biological relevance of fH binding and its possible importance in the pathogenesis of the relapsing-fever spirochetes.
Use of Dipsticks for Rapid Diagnosis of Cholera Caused by
Vibrio cholerae
O1 and O139 from Rectal Swabs
Authors: Bhuiyan N. A. , Qadri Firdausi , Faruque A. S. G. , Malek M. A , Salam M. A. , Nato Farida , Fournier J. M. , Chanteau S. , Sack David A. , Balakrish Nair G.
Published: August 2003

We evaluated the recently developed dipsticks for the rapid detection of
Vibrio cholerae
serotypes O1 and O139 from rectal swabs of hospitalized diarrheal patients after enrichment for 4 h in alkaline peptone water. The sensitivity and specificity of the dipsticks were above 92 and 91%, respectively. The dipsticks represent the first rapid test which has been successfully used to diagnose cholera from rectal swabs, and this would immensely improve surveillance for cholera, especially in remote settings.
Cat Scratch Disease Presenting as Orbital Abscess and Osteomyelitis
Authors: Mirakhur Beloo , Shah Samir S. , Ratner Adam J. , Goldstein Scott M. , Bell Louis M. , Kim Jean O.
Published: August 2003

Ocular manifestations of cat scratch disease are uncommon. The diagnosis is usually made on the basis of increasingBartonella henselaeserum antibody titers. We report a child presenting with orbital abscess and osteomyelitis who was diagnosed with hepatosplenic cat scratch disease by detection ofB. henselaeDNA in the orbital abscess fluid.
Fatal Acute Cellulitis Due to
Neisseria meningitidis
Authors: Cartolano Gian-Luigi , Barbier Christophe , Arnoult Leïla , Simon Didier , Ricome Jean-Louis , Hayon Jan
Published: August 2003

We describe the first fatal evolution of cellulitis due to
Neisseria meningitidis
serogroup Y involving an 85-year-old woman. She presented with an extensive cellulitis of the left side of the face, neck, and thorax and septic shock. In spite of active antibiotic therapy, evolution was rapidly fatal.
Evolutionary dynamics of a satellite DNA in the tiger beetle species pair Cicindela campestris and C. maroccana
Authors: Galián José , Vogler Alfried P
Published: August 2003

Satellite repeat elements are an abundant component of eukaryotic genomes, but not enough is known about their evolutionary dynamics and their involvement in karyotype and species differentiation. We report the nucleotide sequence, chromosomal localization, and evolutionary dynamics of a repetitive DNA element of the tiger beetle species pair Cicindela maroccana and Cicindela campestris. The element was detected after restriction digest of C. maroccana total genomic DNA with EcoRI as a single band and its multimers on agarose gels. Cloning and sequencing of several isolates revealed a consensus sequence of 383 bp with no internal repeat structure and no detectable similarity to any entry in GenBank. Hybridization of the satellite unit to C. maroccana mitotic and meiotic chromosomes revealed the presence of this repetitive DNA in the centromeres of all chromosomes except the Y chromosome, which exhibited only a very weak signal in its short arm. PCR-based tests for this satellite in related species revealed its presence in the sister species C. campestris, but not in other closely related species. Phylogenetic analysis of PCR products revealed well-supported clades that generally separate copies from each species. Because both species exhibit the multiple X chromosome karyotypic system common to Cicindela, but differ in their X chromosome numbers (four in C. maroccana vs. three in C. campestris), structural differences could also be investigated with regard to the position of satellites in a newly arisen X chromosome. We find the satellite in a centromeric position in all X chromosomes of C. maroccana, suggesting that the origin of the additional X chromosome involves multiple karyotypic rearrangements.Key words: chromosome evolution, concerted evolution, Cicindela, Coleoptera, satellite DNA.
Cyto-evolution of Boronia genomes revealed by fluorescent in situ hybridization with rDNA probes
Authors: Shan Fucheng , Yan Guijun , Plummer Julie A
Published: August 2003

The physical location of the 25S26S rDNA sequences was examined in 11 taxa of nine species of Boronia. In diploid species, two rDNA sites were detected in Boronia clavata (2n = 14), Boronia pinnata 'White' (2n = 22), and Boronia chartacea (2n = 32); four in Boronia megastigma (2n = 14) and Boronia denticulata (2n = 18); six in Boronia pinnata 'Pink' (2n = 22); and eight in Boronia molloyae (2n = 16). Eleven sites were found in Boronia heterophylla 'Red' and 'Near White' (2n = 15), but only two active nucleolar organizer regions (NORs) were observed. In polyploid species, Boronia pilosa (2n = 44) had four rDNA sites, while Boronia coerulescens (2n = 72) had six. Most of the rDNA sequences were terminal, but a few were interstitial. There were also differences in signal intensity indicating that the gene copies between and within rDNA sites might be different. The result suggests that considerable chromosome rearrangements have occurred during Boronia cyto-evolution, leading to variation among Boronia taxa in rDNA copy number, site number, and location. These changes together with dysploid reduction during cyto-evolution have made the Boronia genome considerably diverse in chromosome number, genome organization, and chromosome structure.Key words: physical mapping, FISH, chromosome, Rutaceae.
AFLP data and the origins of domesticated crops
Authors: Allaby Robin G , Brown Terence A
Published: August 2003

Amplified fragment length polymorphism (AFLP) datasets have been used to construct neighbor-joining trees from which monophyletic origins for crops such as einkorn wheat, barley, and emmer wheat have been inferred. We simulated several different multiple domestication scenarios for an imaginary cereal crop and examined the resulting domesticated populations. The simulations showed that the population biology aspects of the domestication process can result in independently domesticated populations merging in such a way that a monophyletic origin is erroneously inferred when the resulting population is examined by AFLP genotyping and neighbor-joining analysis. The results bring into question the use of this method to infer the origins of real crops.Key words: AFLPs, agriculture, neighbor-joining, plant domestication.
Mining single-nucleotide polymorphisms from hexaploid wheat ESTs
Authors: Somers Daryl J , Kirkpatrick Robert , Moniwa Mariko , Walsh Andrew
Published: August 2003

Single-nucleotide polymorphisms (SNPs) represent a new form of functional marker, particularly when they are derived from expressed sequence tags (ESTs). A bioinformatics strategy was developed to discover SNPs within a large wheat EST database and to demonstrate the utility of SNPs in genetic mapping and genetic diversity applications. A collection of >90 000 wheat ESTs was assembled into contiguous sequences (contigs), and 45 random contigs were then visually inspected to identify primer pairs capable of amplifying specific alleles. We estimate that homoeologue sequence variants occurred 1 in 24 bp and the frequency of SNPs between wheat genotypes was 1 SNP/540 bp (θ = 0.0069). Furthermore, we estimate that one diagnostic SNP test can be developed from every contig with 1060 EST members. Thus, EST databases are an abundant source of SNP markers. Polymorphism information content for SNPs ranged from 0.04 to 0.50 and ESTs could be mapped into a framework of microsatellite markers using segregating populations. The results showed that SNPs in wheat can be discovered in ESTs, validated, and be applied to conventional genetic studies.Key words: SNP, bioinformatics, EST, genetic mapping.
Modes of reproduction in Australian populations of Hypericum perforatum L. (St. John's wort) revealed by DNA fingerprinting and cytological methods
Authors: Mayo Gwenda M , Langridge Peter
Published: August 2003

Hypericum perforatum L. (St. John's wort) is widely used in homeopathic medicine, but has also become a serious weed in Australia and many other countries. Reproduction in H. perforatum was investigated using markers based on restriction fragment length polymorphism (RFLP) and amplified fragment length polymorphism (AFLP). Between two Australian populations, plants displayed 14 polymorphisms from a total of 22 scorable RFLP markers when genomic DNA was probed with M13 bacteriophage, but individuals within each population exhibited identical RFLP fingerprints. Ninety-four percent of the progeny of four crosses made between the two populations exhibited identical fingerprint and ploidy level to the maternal parent, and probably originated apomictically. Seven seedlings with recombinant RFLP or AFLP fingerprints were found from a total of 121 progeny. Both molecular marker techniques detected the same recombinants from a subset of screened progeny. Cytological analysis showed that the seven recombinants comprised three tetraploids (2n = 4x = 32), three hexaploids (2n = 6x = 48), and one aneuploid (2n  1 = 31), which suggested that the level of normal reduced embryo sacs was only 2.5%. These results are discussed in relation to the management of invasive populations, and the implications for plant breeding and production of St. John's wort for medicinal purposes. Key words: Hypericum perforatum, apomixis, DNA fingerprint, RFLP, AFLP.
A Fuzzy Data Mining Algorithm for Finding Sequential Patterns
Authors: Hu Yi-Chung , Chen Ruey-Shun , Tzeng Gwo-Hshiung , Shieh Jia-Hourng
Published: August 2003

Since fuzzy knowledge representation can facilitate interaction between an expert system and its users, the effective construction of a fuzzy knowledge base is important. Fuzzy sequential patterns described by natural language are one type of fuzzy knowledge representation, and can thus be helpful in building a prototype fuzzy knowledge base. We define that a fuzzy sequence is an ordered list of frequent fuzzy grids, and the length of a fuzzy sequence is the number of frequent fuzzy grids in the frequent fuzzy sequence. Frequent fuzzy grids and frequent fuzzy sequences can be determined by comparing individual fuzzy supports with the user-specified minimum fuzzy support. A fuzzy sequential pattern is just a frequent fuzzy sequence, but it is not contained in any other frequent fuzzy sequence. In this paper, an effective algorithm called the Fuzzy Grids Based Sequential Patterns Mining Algorithm (FGBSPMA) is proposed to generate fuzzy sequential patterns. A numerical example is used to show an analysis of the user visit to websites, demonstrating the usefulness of the proposed algorithm.
Long-Term Efficacy of Globus pallidus Stimulation for the Treatment of Parkinson’s Disease
Authors: Lyons Kelly E. , Wilkinson Steven B. , Tröster Alexander I. , Pahwa Rajesh
Published: August 2003

Objective: To determine the long-term efficacy and safety of globus pallidus internus (GPi) stimulation for Parkinson’s disease (PD). Background: We previously reported 3-month data for 5 patients who underwent GPi stimulation for PD. We now report long-term data on these 5 patients and 4 additional patients. Methods: Nine PD patients, 5 men and 4 women, with an average age of 49 years and disease duration of 10 years, underwent GPi stimulation. Six patients had staged bilateral implants and 3 patients had unilateral implants. The mean follow-up was 48.5 months. All patients were evaluated with the Unified Parkinson’s Disease Rating Scale (UPDRS) and completed 2-day diaries before and after surgery. Results: There was a 21% improvement in UPDRS Part II (activities of daily living; ADL) scores and a 37% improvement in UPDRS Part III (motor) scores when the longest follow-up in the ‘stimulation-on/medication-off’ state was compared to the ‘medication-off’ state at baseline. The UPDRS Part II (ADL) scores improved by 30% and the UPDRS Part III (motor) scores improved by 39% when the longest follow-up in the ‘stimulation-on/mediation-on’ state was compared to the ‘medication-on’ state at baseline. As measured by patient diaries, ‘on’ time increased from 25 to 59% and ‘on with dyskinesia’ decreased from 42 to 15%. Surgical- and device-related complications included transient hemiparesis in the operating room, postoperative seizures, and implantable pulse generator and lead problems. There were seven device-related events requiring additional surgical procedures. Conclusions: GPi stimulation continues to be effective for the long-term treatment of the disabling symptoms of PD; however, the physician and patient should be aware that device-related problems are not uncommon and additional surgery may be necessary.
Gamma Knife Radiosurgery for Brain Metastases: Do Patients Benefit from Adjuvant External-Beam Radiotherapy? An 18-Month Comparative Analysis
Authors: Jawahar Ajay , Willis Brian K. , Smith Donald R. , Ampil Federico , Datta Ratna , Nanda Anil
Published: August 2003

Objective: To analyze 18 months of results of gamma knife stereotactic radiosurgery in the treatment of brain metastases and determine factors affecting outcome by examining the effectiveness of additional external-beam radiotherapy (XRT). Materials and Methods: Between January 2000 and September 2001, 61 patients with 103 tumors diagnosed as cerebral metastases were treated with gamma knife. Mean patient age was 57 years (range = 36–81). Lung carcinoma (55.7%) was the most common primary cancer, followed by melanoma (14.8%) and breast carcinoma (11.5%). Mean KPS of the patients was 70 (range = 50–90). Twenty-seven patients had solitary metastases while 34 had multiple tumors. Forty-three patients (59 tumors in total) received only radiosurgery, while 18 patients (44 tumors in total) had prior XRT. Tumor volume ranged from 0.5 to 33 cm3 (mean = 9.74 cm3). Mean marginal dose prescription to the tumor was 15 Gy (range = 11–21 Gy). Results: Median follow-up was 11 months. Twenty-one patients (34.4%) were alive at last follow-up and 40 (65.6%) had died. Seventeen deaths (42.5%) were reported to be due to progressive brain disease, while 23 deaths (57.5%) were due to uncontrolled primary. Control of the treated lesions was achieved in 45 patients (73.8%) and 84 tumors (81.6%). Mean overall survival of the patients is 8 months (range = 1–19 months). The actuarial 12-month tumor control rate using the Kaplan-Meier method for this series is 68.2 ± 0.06%. Results of the log-rank test revealed that younger age (<55 years), small tumor volume (<10 cm3), and increasing tumor dose (>15 Gy) correlated with improved brain disease-free survival (p < 0.05). Overall survival, local tumor control rate and the freedom from brain disease period (based on the appearance of new brain tumors after radiosurgery) were analyzed separately for the groups receiving radiosurgery alone and those with prior XRT to detect any additional benefit of XRT. No statistically significant difference was found between the two groups for any of the considered outcomes. Conclusion: Gamma knife stereotactic radiosurgery is a safe and effective treatment option for patients with cerebral metastases. It provides survival benefits and improves quality of life by achieving excellent control of the brain disease, irrespective of patients’ age or number of brain tumors. The addition of XRT in younger patients with small brain metastases does not improve survival and/or control of the brain disease.
Biochemical Characterization of the First Essential Two-Component Signal Transduction System from Staphylococcus aureus and Streptococcus pneumoniae
Authors: Clausen Valerie A. , Bae Weonhye , Throup John , Burnham Martin K.R. , Rosenberg Martin , Wallis Nicola G.
Published: August 2003

The yycFG two-component signal transduction system (TCSTS) has been shown to be essential to the viability of several gram-positive bacteria. However, the function of the gene pair remains unknown. Interestingly, while both components are essential to Staphylococcus aureus and Bacillus subtilis, only the response regulator (yycF) is essential to Streptococcus pneumoniae. To study this essential TCSTS further, the S. pneumoniae and S. aureus truncated YycG histidine kinase and full-length YycF response regulator proteins were characterized at a biochemical level. The recombinant proteins from both organisms were expressed in Escherichia coli and purified. The YycG autophosphorylation activities were activated by ammonium. The apparent Km (ATP) of S. aureus YycG autophosphorylation was 130 µM and S. pneumoniae was 3.0 µM. Each had similar kcat values of 0.036 and 0.024 min–1, respectively. Cognate phosphotransfer was also investigated indicating different levels of the phosphorylated YycG intermediates during the reaction. The S. pneumoniae YycG phosphorylated intermediate was not detectable in the presence of its cognate YycF, while phosphorylated S. aureus YycG and YycF were detected concurrently. In addition, noncognate phosphotransfer was demonstrated between the two species. These studies thoroughly compare the essential YycFG TCSTS from the two species at the biochemical level and also establish methods for assaying the activities of these antibacterial targets.
The Impact of the Day of the Week and Month of Admission on the Length of Hospital Stay in Stroke Patients
Authors: Schmidt Wolf-Peter , Taeger Dirk , Buecker-Nott Hans-Joachim , Berger Klaus
Published: August 2003

Background: Length of hospital stay (LOS) in many diseases is determined by patient-related sociodemographic and clinical factors but also by the management of treatment and care. We examined the influence of the day of the week (DW) and month of admission on LOS in stroke patients. Methods: We used data from a large regional stroke registry in the northwest of Germany for the years 2000 and 2001. The registry is based on a standardised data assessment in participating hospitals, including patient-related sociodemographic and clinical items, diagnostic and treatment procedures, stroke classification, as well as admission and discharge dates. Data management and analysis were done centrally. Results: A total of 11,942 patients from 42 hospital departments were included. Median LOS was 13 days for ischemic stroke and 8 days for TIA. Patients with ischaemic stroke admitted on Tuesdays or Wednesdays had the longest LOS. We observed a significantly shorter LOS in patients discharged towards the weekend, both for ischaemic stroke and TIA, independent of age, gender and symptom severity. Discharges were predominantly done on Fridays (25%), transferrals to rehabilitation centres most often on Thursdays (28%) in neurologic departments but on Tuesdays in departments of internal medicine. LOS according to the month of admission revealed a biphasic curve for both ischaemic stroke and TIA, with shortest LOS in July and December. Conclusions: Discharge patterns on weekends contribute to differences in LOS for the various DWs of admission. Changes in discharge organisation and improved co-operation with rehabilitation centres may shorten LOS. Differences in LOS between the months show considerable seasonal variability.
Coxsackievirus B3 Induces Apoptosis in the Early Phase of Murine Myocarditis: A Comparative Analysis of Cardiovirulent and Noncardiovirulent Strains
Authors: Joo Chul Hyun , Hong Hea Nam , Kim Eun Ok , Im Jin Ok , Yoon Seung-Yong , Ye Jeong Sook , Moon Mi Sun , Kim Donghou , Lee Heuiran , Kim Yoo Kyum
Published: August 2003

Objective: To investigate the relationship between enteroviral infection and myocardial tissue apoptosis during the development of viral myocarditis in a murine model. Methods: C3H/HeJ mice were inoculated with two strains of coxsackievirus B3, specifically CVB3 (cardiovirulent Nancy strain) and CVB3/0 (noncardiovirulent strain). Mice were sacrificed at 4, 7 and 10 days postinfection (p.i.). Hearts were removed, and plaque assays and RT-PCR were performed to detect the presence of viruses. Myocardial tissue sections were additionally evaluated by hematoxylin and eosin staining for inflammation, VP1 and Bax immunohistochemical staining for detection of virus and Bax expression, and TUNEL and Apostain for localization of apoptosis. Results: CVB3 replicated to significantly higher titers than CVB3/0 at all time points. Histopathological analyses revealed significant inflammatory changes at all time points in CVB3-infected mice, in contrast to minimal changes in CVB3/0-infected mice. TUNEL and Apostain assays of myocardial tissues from mice infected with CVB3 disclosed maximum apoptotic lesions at 4 days p.i. and to a lesser extent at 7 and 10 days p.i. Moreover, CVB3-infected myocardial tissues displayed significantly enhanced Bax expression at 4 days p.i., and lesions overlapped with VP1-stained areas. Conclusions: These data indicate that (1) the cardiovirulent strain CVB3 induces more severe inflammation and apoptosis than the noncardiovirulent CVB3/0 strain, (2) viral replication is localized in inflammatory and apoptotic lesions in myocardial tissues, (3) apoptotic changes are observed in the early stages of myocarditis and (4) Bax may be associated with the apoptosis process in CVB3-induced myocarditis.
Pleiotropy and Heterogeneity in the Expression of Atherogenic Lipoproteins: The IRAS Family Study
Authors: Hokanson John E. , Langefeld Carl D. , Mitchell Braxton D. , Lange Leslie A. , Goff Jr. David C. , Haffner Steven M. , Saad Mohammad F. , Rotter Jerome I.
Published: August 2003

Objective:  Dyslipidemia is an important determinant of coronary disease. Phenotypic correlations between atherogenic lipids are well established, but the contribution of common genetic influences is less clear. Methods: This study investigates the pair-wise genetic (ρg) and environmental (ρe) correlations between apoB, LDL-C, HDL-C, and triglyceride (Tg) from Hispanic and African American families of the IRAS Family Study. Results: Heritability estimates (hcirc;2) indicate significant genetic effects on apoB (hcirc;2 = 0.46 ± 0.05), LDL-C (hcirc;2 = 0.40 ± 0.05), HDL-C (hcirc;2 = 0.47 ± 0.05), and Tg (hcirc;2 = 0.35 ± 0.05) (all p < 0.001). Genetic and environmental correlations were strong for apoB – LDL-C (ρg = 0.87, ρe = 0.84), apoB – Tg (ρg = 0.38, ρe = 0.65), and HDL-C – Tg (ρg = –0.42, ρe = –0.46). Environmental correlations were strong for apoB – HDL-C (ρe = –0.40), LDL-C – HDL-C (ρe = –0.24), and Tg – LDL-C (ρe = 0.33) with weak genetic correlations for these pairs (ρg = –0.09, 0.10, 0.09 respectively). Conclusions: These results suggest multiple pathways leading to atherogenic dyslipidemia. There are common genetic and environmental influences contributing to variations in apoB and LDL-C as well as apoB and Tg. In addition, the inverse relation between Tg and HDL-C appears to have both genetic and environmental basis. Identifying genes involved in atherogenic dyslipidemia will require careful dissection of the genetic architecture of these pathways.
Sex-Specific Mutation Rates for X-Linked Disorders: Estimation and Application
Authors: Flodman Pamela , Hodge Susan E.
Published: August 2003

Emerging human molecular data are adding to our knowledge about the frequency and pattern of genetic mutations. This not only gives important insight into the biological processes underlying mutation, but also provides data which must be incorporated in the clinical setting. An example is the assumption of equal mutation probability in the male and female germ lines. This is a key assumption in Bayesian risk calculation for families segregating an X-linked recessive disorder. For some disorders, data are now available that demonstrate that the mutation probability in males differs from that in females. In this paper, we review the estimation of the male-female mutation rate ratio, including the construction of confidence intervals, and apply sex-specific mutation rates to carrier risk calculation in a variety of pedigree structures. In several instances, the difference in risk is substantial.
Adhesion of Endothelial Cells to NOV Is Mediated by the Integrins αvβ3 and α5β1
Authors: Ellis Peter D. , Metcalfe James C. , Hyvönen Marko , Kemp Paul R.
Published: August 2003

NOV is a member of the CCN family of matricellular proteins. We have shown previously that Nov is strongly expressed by vascular smooth muscle cells (VSMCs) of the rat carotid artery. However, 7 days after injury, Nov expression is down-regulated, except near the luminal surface of the developing intima, where it is strongly expressed. These data suggested that NOV might be involved in the regulation of endothelial cell adhesion. NOV promoted the adhesion of human umbilical vein endothelial cells (HUVECs), which was abolished by anti-NOV antibody. HUVEC adhesion to NOV required divalent cations and was inhibited by GRGDS peptide, implicating integrins in the adhesion mechanism. Monoclonal antibodies (mAbs) against αvβ3 inhibited adhesion of HUVECs to NOV, and NOV was shown to bind to αvβ3. Anti-α5β1 mAbs also inhibited HUVEC adhesion to NOV, but adhesion via α5β1 was mediated by fibronectin. HUVEC adhesion to NOV caused intracellular signalling, as evidenced by increased phosphotyrosine content of focal adhesion kinase. Together with evidence that Nov expression in a variety of tissues is restricted to blood vessels containing VSMCs, these data are consistent with a role for NOV in endothelial cell adhesion in vascular homeostasis and in the response to injury.
Intermittent High-Permeability Hemofiltration Modulates Inflammatory Response in Septic Patients with Multiorgan Failure
Authors: Morgera Stanislao , Haase Michael , Rocktäschel Jens , Böhler Torsten , Vargas-Hein Ortrud , Melzer Christoph , Krausch Dietmar , Kox Wolfgang J. , Baumann Gerd , Beck Werner , Göhl Hermann , Neumayer Hans-H.
Published: August 2003

Background/Aim: Continuous venovenous hemofiltration with high-permeability hemofilters is a novel approach in the adjuvant therapy of septic patients. High-permeability hemofilters are characterized by an increased pore size which facilitates the filtration of inflammatory mediators. The present study examines whether intermittent high-permeability hemofiltration has an immunomodulatory effect on polymorphonuclear leukocytes and mononuclear cells. Methods: Twenty-eight septic patients with acute renal failure were randomly allocated to either receive intermittent high-permeability or conventional hemofiltration. Intermittent high-permeability hemofiltration consisted of a daily 12-hour course of high-permeability hemofiltration alternated by conventional hemofiltration. For high-permeability hemofiltration, a newly developed high-flux polyamide membrane (P2SH) with a nominal cutoff point of 60 kD was used. For conventional hemofiltration a high-flux polyamide hemofilter (Polyflux 11S, cutoff point 30 kD) was used. Results: The polymorphonuclear leukocyte phagocytosis activity before starting hemofiltration was almost double the rate of healthy controls in both groups (p < 0.001). The phagocytosis rate decreased significantly during the course of intermittent high-permeability hemofiltration (p < 0.05), whereas the values remained unchanged in the conventional hemofiltration group. Incubation of high-permeability filtrates with blood from healthy donors resulted in a significant induction of phagocytosis (p < 0.001), whereas conventional filtrates had no phagocytosis-stimulating effects. In addition, incubation of healthy-donor mononuclear cells with high-permeability but not conventional filtrates resulted in a significant tumor necrosis factor alpha release (p < 0.001). Conclusions: Intermittent high-permeability hemofiltration is a novel extracorporeal elimination modality which exhibits immunomodulatory effects on leukocytes, attenuating polymorphonuclear neutrophil phagocytosis. Further studies are necessary to elucidate whether these effects translate in a clinical improvement in patients suffering from sepsis.
Vasodilator Reactivity to Calcitonin Gene-Related Peptide Is Increased in Mesenteric Arteries of Rats during Early Pregnancy
Authors: van Eijndhoven H.W.F. , van der Heijden O.W.H. , Fazzi G.E. , Aardenburg R. , Spaanderman M.E.A. , Peeters L.L.H. , De Mey J.G.R.
Published: August 2003

The objective of the present study was to determine the effect of early pregnancy on the sensitivity to, and endogenous production of calcitonin gene-related peptide (CGRP). Contractile responses of arteries of 10-day pregnant and nonpregnant rats were studied in myographs. During contractions induced by 40 mmol/l K+, exogenous CGRP elicited an approximately 30% stronger relaxation in mesenteric arteries in pregnancy, an effect not seen in renal and uterine arteries. Capsaicin treatment during K+-induced contractions caused a persistent potentiation of the contractile response in mesenteric arteries, indicating that K+ stimulates the endogenous release of CGRP. This potentiation was similar in the pregnant and nonpregnant state (+81 ± 23% and +82 ± 23%, respectively), suggesting no effect of pregnancy on the endogenous CGRP release. The latter was paralleled by comparable CGRP content in the arteries of both groups, indicating similar tissue CGRP availability. The results of this study support the concept that early pregnancy is associated with a rise in the vascular sensitivity to CGRP in selected areas of the vascular bed without concomitant increase in the vascular CGRP production and release.
Double Resection for Patients with Pancreatic Cancer and a Second Primary Renal Cell Cancer
Authors: Alexakis N. , Bosonnet L. , Connor S. , Ellis I. , Sutton R. , Campbell F. , Hughes M. , Garvey C. , Neoptolemos J.P.
Published: August 2003

Background: Reports of synchronous or metachronous double kidney-pancreas cancers are very rare. Methods: We present 2 patients with renal cell carcinoma and synchronous (1 patient) or metachronous (1 patient) primary pancreatic ductal adenocarcinoma. The patients underwent resection for both cancer types with a worthwhile outcome. Results: The appearance of different primaries in an individual may indicate a genetic predisposition to different neoplasms. The study of double primary cancers is important because it might provide understanding of a shared genetic basis of different solid tumors. Conclusions: The association between these two cancers demands more detailed epidemiological and molecular investigation. From a clinical viewpoint a resectional policy is recommended.
Analgesic Efficacy of Etoricoxib in Primary Dysmenorrhea: Results of a Randomized, Controlled Trial
Authors: Malmstrom Kerstin , Kotey Paul , Cichanowitz Nicole , Daniels Stephen , Desjardins Paul J.
Published: August 2003

Objective: To determine the efficacy of etoricoxib in the treatment of primary dysmenorrhea. Methods: Seventy-three women were randomly assigned to receive single oral doses of etoricoxib 120 mg, placebo, or naproxen sodium 550 mg at the onset of moderate to severe pain associated with menses. During 3 consecutive menstrual cycles in this double-blind, 3-period, crossover study, pain intensity and pain relief were assessed over the 24-hour period following dosing, and global ratings of therapy were made at 8 and 24 h after dosing. Tolerability was assessed by spontaneous reports of adverse experiences. Results: Etoricoxib 120 mg provided analgesic efficacy superior to placebo for the primary endpoint, total pain relief over 8 h (TOPAR8, p < 0.001), and for all secondary endpoints (p < 0.050). The analgesic effect of etoricoxib 120 mg over the first 8 h was similar to that of naproxen sodium 550 mg. All treatments were well tolerated. Conclusions: Etoricoxib 120 mg provided rapid and sustained analgesia that was superior to placebo and similar to that of naproxen sodium 550 mg.
GP130, the shared receptor for the LIF/IL6 cytokine family in the mouse,is not required for early germ cell differentiation, but is required cell-autonomously in oocytes for ovulation
Authors: Molyneaux Kathleen A. , Schaible Kyle , Wylie Christopher
Published: August 2003

GP130 is the shared receptor for members of the IL6 family of cytokines. Members of this family have been shown to enhance the survival of migratory(E10.5) or postmigratory (E12.5) murine primordial germ cells (PGCs) in culture; however, it is uncertain what role these cytokines play during PGC development in vivo. We have examined PGC numbers in E13.5 GP130-deficient mouse embryos and found that males exhibited a slight decrease in PGC numbers;females were normal. Also, we used the Cre-loxP system to inactive GP130 specifically in germ cells and found that this resulted in a fertility defect in females. These animals were found to have a slight reduction in the number of primary follicles and a major defect in ovulation. This data suggests that GP130 is required in female germ cells for their normal function, but is dispensable in male germ cells.
Vaginal cytology and vulvar swelling as indicators of impending estrus and ovulation in the giant panda (Ailuropoda melanoleuca)
Authors: Durrant Barbara S. , Ann Olson Mary , Amodeo Dianne , Anderson Autumn , Russ Kara D. , Campos‐Morales Rogelio , Gual‐Sill Fernando , Ramos Garza Juan
Published: August 2003

The giant panda is critically endangered in its native habitat, which is continually depreciated by the pressure of human population growth, and the small captive population is not yet self‐sustaining. Female giant pandas are seasonally monoestrous with spontaneous ovulation. Precise monitoring of the estrous cycle to pinpoint the time of ovulation is critical for the success of both timed mating and artificial insemination. Vaginal cytology is a useful adjunct to endocrinological data in describing the estrous cycle in many species, including domestic carnivores; however, it has not been used effectively in the giant panda. In this study, vaginal swabs were obtained from four nulliparous females ranging in age from 6 to 14 years. Cells were stained with Papanicolaou stain and classified as basophils, intermediate cells, or superficial cells based on traditional criteria. In addition, cell color changes from the basophilic through the acidophilic and keratinized stages were recorded. The size and color of the vulva of a single female was scored daily on a scale of 0–3, with anestrus characterized as 0. The day of ovulation was retrospectively designated as day 0 based on urinary estrogen concentrations, behavior, and morphological indicators. The vulva began to swell between days –11 and –9, with peak tumescence on the day of ovulation. The percentage of superficial cells began to increase by day –8 to –7, and peaked between days –2 and +2. A chromic shift occurred on day –8, when the majority of vaginal cells changed from basophilic to acidophilic without any accompanying nuclear or cytoplasmic changes. A second chromic shift was observed on day –2, when keratinized cells replaced acidophilic cells as the majority of vaginal cells. By day +4, all parameters had returned to baseline except vulvar swelling, which slowly subsided over the next several days. These data show the correlation of trichrome‐stained vaginal cytology with other measured indicators of estrus in the giant panda, and its predictive value in determining the time of ovulation. Zoo Biol 22:313–321, 2003. © 2003 Wiley‐Liss, Inc.
Use of adhesive surgical tape with the absorbable continuous subcuticular suture
Authors: Kolt Jeremy D.
Published: August 2003

Background:  The absorbable continuous subcuticular suture is frequently used to close surgical incisions where the aim is healing by primary intention. A form of adhesive surgical tape is commonly also placed over the wound but this combination closure seems to have its development based on anecdotal, rather than experimental evidence. The present study reviews the scientific literature on the development of sutureless wound closure and presents the current evidence for the use of combination wound closure.

Methods:  Review was undertaken of the medical literature using the PubMed Internet database and cross‐referencing major ­articles on the subject. The following combinations of key words were searched: skin closure, wound closure, suture technique, sutureless, adhesive tape, op‐site, staples, subcuticular suture, complication, infection and scars.

Results:  Taped closure alone has advantages of lower wound infection rates and greater wound tensile strength, but disadvantages of epidermal reaction, skin edge inversion, doubtful safety and time required for meticulous surgical technique. The use of the continuous absorbable subcuticular suture allows accurate skin edge approximation, which increases the safety margin. The combination closure has a slightly superior cosmetic result to sutureless techniques but no study has been performed to compare the results of combination subcuticular suture and tape, with tape or subcuticular suture alone.

Conclusions:  There is no evidence in the scientific literature to justify or support the practice of closing a surgical wound with both subcuticular suture and adhesive surgical tape.