bluuebunny/crossref_metadata_2025_split huggingface dataset:
The rise of buprenorphine prescribing in England: analysis of NHS regional data, 2001–03
Authors: De Wet Cornelis J. , Reed Laurence J. , Bearn Jennifer 
Published: March 2005

ABSTRACTAims  Since its launch in the prescribing market in 1999 for the treatment of opiate dependence, buprenorphine has rapidly become established as an alternative to methadone treatment in the United Kingdom. In the absence of evidence of its clinical superiority over methadone, and given its high relative cost, we sought to examine the impact of buprenorphine availability on opiate treatment services in England.Methods  Quarterly buprenorphine and methadone community prescription figures were obtained for 28 Strategic Health Authorities (SHAs) in England, for the 2‐year period September 2001 to September 2003. Rates of buprenorphine prescribing (as proportion of all opiate prescriptions) were examined over time by number of prescriptions and net ingredient cost.Results  Buprenorphine prescription rates increased disproportionately to methadone in all 28 SHAs. By the end of 2003 the number of buprenorphine prescriptions had increased to 23% of all opiate prescriptions, but accounted for 45% of opiate prescription costs in England. Buprenorphine prescribing rates varied substantially across different regions.Conclusions  Buprenorphine prescribing has increased dramatically and represents a disproportionately large fraction of community opiate prescribing costs. The marked regional variation suggests the need for further research and the development of national guidelines to support rational prescribing and equitable access to treatment.
Expressed sequence tags from Diabrotica virgifera virgifera midgut identify a coleopteran cadherin and a diversity of cathepsins
Authors: Siegfried B. D. , Waterfield N. , Ffrench‐Constant R. H. 
Published: March 2005

AbstractThe Western corn rootworm is the major pest of corn in the USA and has recently become the target for insect‐resistant transgenic crops. Transgenic crops have switched the focus for identifying insecticide targets from the insect nervous system to the midgut. Here we describe a collection of 691 sequences from the Western corn rootworm midgut, 27% of which predict proteins with no matches in current databases. Of the remaining sequences, most predict proteins with either catalytic (62%) or binding (19%) functions, as expected for proteins expressed in the insect midgut. The utility of this approach for the identification of targets for novel toxins is demonstrated by analysis of the first coleopteran cadherin gene, a putative Bt receptor, and a large class of cysteine‐proteases, the cathepsins.
In Vitro Susceptibility Testing ofBorrelia burgdorferiSensu Lato Isolates Cultured from Patients with Erythema Migrans before and after Antimicrobial Chemotherapy
Authors: Hunfeld Klaus-Peter , Ruzic-Sabljic Eva , Norris Douglas E. , Kraiczy Peter , Strle Franc 
Published: March 2005

ABSTRACTClinical treatment failures have been reported to occur in early Lyme borreliosis (LB) for many suitable antimicrobial agents. Investigations of possible resistance mechanisms of theBorrelia burgdorfericomplex must analyze clinical isolates obtained from LB patients, despite their receiving antibiotic treatment. Here, borrelial isolates obtained from five patients with erythema migrans (EM) before the start of antibiotic therapy and again after the conclusion of treatment were investigated. The 10 isolates were characterized by restriction fragment length polymorphism analysis and plasmid profile analysis and subjected to susceptibility testing against a variety of antimicrobial agents including those used for initial chemotherapy. Four out of five patients were infected by the same genospecies (Borrelia afzelii,n= 3;Borrelia garinii,n= 1) at the site of the EM lesion before and after antimicrobial therapy. In one patient the genospecies of the initial isolate (B.afzelii) differed from that of the follow-up isolate (B.garinii). No significant changes in the in vitro susceptibilities became obvious for corresponding clinical isolates before the start and after the conclusion of antimicrobial therapy. This holds true for the antimicrobial agents used for specific chemotherapy of the patients, as well as for any of the additional agents tested in vitro. Our study substantiates borrelial persistence in some EM patients at the site of the infectious lesion despite antibiotic treatment over a reasonable time period. Borrelial persistence, however, was not caused by increasing MICs or minimal borreliacidal concentrations in these isolates. Therefore, resistance mechanisms other than acquired resistance to antimicrobial agents should be considered in patients with LB resistant to treatment.
Is In Vitro Antibiotic Combination More Effective than Single-Drug Therapy against Anthrax?
Authors: Athamna Abed , Athamna Muhammad , Nura Aburashed , Shlyakov Eli , Bast Darrin J. , Farrell David , Rubinstein Ethan 
Published: March 2005

ABSTRACT

Antibiotic combinations are used to enhance antibacterial efficacy and to prevent the development of resistance. We have tested a possible synergistic effect of several antibacterial combinations on
Bacillus anthracis
. The in vitro activities of antibiotic combinations against two strains of
B. anthracis
, strain Sterne and the Russian anthrax vaccine strain STi, were tested by the fractional inhibitory concentration (FIC) method, derived from the MICs of the agents in combination, and by measuring the rate of bacterial killing over time by several antibiotic combinations. The FIC results showed that synergism against both
B. anthracis
strains was observed only with the combination of rifampin and clindamycin. The telithromycin-amoxicillin combination showed synergism against strain Sterne only. All other combinations were either indifferent or antagonistic. The results of the bacterial time-kill study demonstrated indifferent effects for all combinations. These in vitro results demonstrate the difficulties in obtaining synergistic combinations of antibiotics against
B. anthracis
.
Testing of the In Vitro Susceptibilities ofMadurella mycetomatisto Six Antifungal Agents by Using the Sensititre System in Comparison with a Viability-Based 2,3-Bis(2-Methoxy-4-Nitro-5-Sulfophenyl)-5- [(Phenylamino)Carbonyl]-2H-Tetrazolium Hydroxide (XTT) Assay and a Modified NCCLS Method
Authors: van de Sande Wendy W. J. , Luijendijk Ad , Ahmed Abdalla O. A. , Bakker-Woudenberg Irma A. J. M. , van Belkum Alex 
Published: March 2005

ABSTRACTThe in vitro susceptibilities of 36 clinical isolates ofMadurella mycetomatis, the prime agent of eumycetoma in Africa, to ketoconazole, itraconazole, fluconazole, voriconazole, amphotericin B, and flucytosine were determined by the Sensititre YeastOne system. This system appeared to be a rapid and easy test, and by use of hyphal suspensions it generated results comparable to those of a modified NCCLS method. After 10 days of incubation, the antifungal activities of ketoconazole (MIC at which 90% of isolates were inhibited [MIC90], 0.125 μg/ml), itraconazole (MIC90, 0.064 μg/ml), and voriconazole (MIC90, 0.125 μg/ml) appeared superior to those of fluconazole (MIC90, 128 μg/ml) and amphotericin B (MIC90, 1 μg/ml), with MICs in the clinically relevant range. All isolates were resistant to flucytosine (all MICs above 64 μg/ml). Based on the relatively broad range of MICs obtained for the antifungal agents, routine testing ofM. mycetomatisisolates for susceptibility to antifungal agents seems to be relevant to adequate therapeutic management.
Development of Cell-Based Assays for In Vitro Characterization of Hepatitis C Virus NS3/4A Protease Inhibitors
Authors: Chung Victoria , Carroll Anthony R. , Gray Norman M. , Parry Nigel R. , Thommes Pia A. , Viner K. Claire , D'Souza Eric A. 
Published: March 2005

ABSTRACT

A recombinant vaccinia virus, expressing the NS3-to-NS5 region of the N clone of hepatitis C virus (HCV), was generated and utilized both in a gel-based assay and in an enzyme-linked immunosorbent assay (ELISA) to evaluate the pyrrolidine-5,5-
trans
-lactams, a series of inhibitors of the HCV NS3/4A protease. The absolute levels of processed, mature HCV nonstructural proteins in this system were found to decrease in the presence of the
trans-
lactams. Monitoring of this reduction enabled end points and 50% inhibitory concentrations to be calculated in order to rank the active compounds according to potency. These compounds had no effect on the transcription or translation of the NS3-5 polyprotein at concentrations shown to inhibit NS3/4A protease, and they were shown to be specific inhibitors of this protease. The ELISA, originally developed using the vaccinia virus expression system, was modified to utilize Huh-7 cells containing an HCV replicon. Results with this assay correlated well with those obtained with the recombinant vaccinia virus assays. These results demonstrate the utility of these assays for the characterization of NS3/4A protease inhibitors. In addition, inhibitors of other viral targets, such as polymerase and helicase, can be evaluated in the context of the replicon ELISA.
Silencing of Glycopeptide Resistance in
Enterococcus faecalis
BM4405 by Novobiocin
Authors: Patiño Lorena Abadía , Chippaux Marc , Courvalin Patrice , Périchon Bruno 
Published: March 2005

ABSTRACT

Enterococcus faecalis
BM4405-1, a susceptible derivative of the VanE-type vancomycin-resistant
E. faecalis
strain BM4405, was obtained after growth in the presence of novobiocin, an inhibitor of the GyrB subunit of DNA gyrase. In contrast to findings for BM4405, UDP-MurNAc-
l
-Ala-γ-
d
-Glu-
l
-Lys-
d
-Ala-
d
-Ala (pentapeptide[
d
-Ala]) was the only peptidoglycan precursor found in BM4405-1, and no VanXY
E
d
,
d
-peptidase or VanT serine racemase activities were detected in that strain, even after induction by subinhibitory concentrations of vancomycin. Sequencing of the
vanE
operon of BM4405-1 revealed two mutations leading to substitutions in VanE (D200N) and in the C-terminal amino acid of VanR
E
(Y225F). Cloning of the
vanE
,

vanXY
E

, and

vanT
E

genes of BM4405-1 into the susceptible
E. faecalis
strain JH2-2 conferred resistance to vancomycin, indicating that the mutation in
vanE
was not responsible for susceptibility. Transcriptional analysis of the
vanE
operon in BM4405 by quantitative reverse transcription-PCR indicated that novobiocin did not affect the expression level of the
vanE
operon. Sequencing of the
gyrB
gene of BM4405-1 revealed a mutation responsible for substitution of a residue (K337Y) required for ATPase activity and thus implicated in DNA supercoiling. Cloning of the
gyrB
gene of BM4405 restored vancomycin resistance to BM4405-1. Taken together, these data suggest that alteration of DNA supercoiling following a mutation in GyrB was responsible for lack of expression of the
vanE
operon and thus for vancomycin susceptibility in BM4405-1.
Acquisition of Resistance to Carbapenems in Multidrug-Resistant Clinical Strains of
Acinetobacter baumannii
: Natural Insertional Inactivation of a Gene Encoding a Member of a Novel Family of β-Barrel Outer Membrane Proteins
Authors: Mussi María A. , Limansky Adriana S. , Viale Alejandro M. 
Published: March 2005

ABSTRACT

The outer membrane proteins responsible for the influx of carbapenem β-lactam antibiotics in the nonfermentative gram-negative pathogen
Acinetobacter baumannii
are still poorly characterized. Resistance to both imipenem and meropenem in multidrug-resistant clinical strains of
A. baumannii
is associated with the loss of a heat-modifiable 29-kDa outer membrane protein, designated CarO. The chromosomal locus containing the
carO
gene was cloned and characterized from different clinical isolates. Only one
carO
copy, present in a single transcriptional unit, was found in the
A. baumannii
genome. The
carO
gene encodes a polypeptide of 247 amino acid residues with a typical N-terminal signal sequence and a predicted transmembrane β-barrel topology. Its absence from different carbapenem-resistant clinical isolates of
A. baumannii
resulted from the disruption of
carO
by distinct insertion elements. The overall data thus support the notion that CarO participates in the influx of carbapenem antibiotics in
A. baumannii
. Moreover, database searches identified the presence of
carO
homologs only in species of the genera
Acinetobacter
,
Moraxella
, and
Psychrobacter
, disclosing the existence of a novel family of outer membrane proteins restricted to the family
Moraxellaceae
of the class γ
-Proteobacteria
.
CTX-M-10 Linked to a Phage-Related Element Is Widely Disseminated among
Enterobacteriaceae
in a Spanish Hospital
Authors: Oliver Antonio , Coque Teresa M. , Alonso Diana , Valverde Aránzazu , Baquero Fernando , Cantón Rafael 
Published: March 2005

ABSTRACT

CTX-M-10 has been widely disseminated among multiple clones of several species of
Enterobacteriaceae
, harboring seemingly different plasmids, for over a decade in Ramón y Cajal University Hospital, Madrid, Spain. Cloning and sequencing of a 12.2-kb DNA fragment from plasmid pRYCE21 from
Klebsiella pneumoniae
strain KP4aC revealed a novel phage-related element immediately upstream of
bla
CTX-M-10
conserved among different CTX-M-10-producing strains. This is the first report showing an extended-spectrum-β-lactamase gene linked to a phage-related element.
Occurrence of CTX-M-3, CTX-M-15, CTX-M-14, and CTX-M-9 Extended-Spectrum β-Lactamases in
Enterobacteriaceae
Clinical Isolates in Korea
Authors: Kim Jungmin , Lim Yu-Mi , Jeong Young-Sook , Seol Sung-Yong 
Published: March 2005

ABSTRACT

Among 603 isolates of
Enterobacteriaceae
collected between June and November 2003 from three university hospitals within Korea,
bla
CTX-M-3
,
bla
CTX-M-15
,
bla
CTX-M-14
, and
bla
CTX-M-9
were detected in 41 isolates of species from five different genera of
Enterobacteriaceae
,
Escherichia coli
,
Klebsiella pneumoniae
,
Citrobacter freundii
,
Enterobacter
spp., and
Serratia marcescens
.
Capsule Homology Does Not Increase the Frequency of Transformation of Linked Penicillin Binding Proteins PBP 1a and PBP 2x in
Streptococcus pneumoniae
Authors: Trzciński Krzysztof , MacNeil Adam , Klugman Keith P. , Lipsitch Marc 
Published: March 2005

ABSTRACT

Penicillin resistance is mainly confined to a limited number of
Streptococcus pneumoniae
serotypes. Given linkage between the capsular biosynthesis locus and two penicillin binding proteins, we tested whether capsule homology increases transformation rates of penicillin resistance. Transformation rates in homologous donor-recipient pairs were no higher than expected, falsifying this hypothesis.
Sensititre YeastOne Caspofungin Susceptibility Testing of
Candida
Clinical Isolates: Correlation with Results of NCCLS M27-A2 Multicenter Study
Authors: Cantón Emilia , Pemán Javier , Gobernado Miguel , Álvarez Elena , Baquero Fernando , Cisterna Ramón , Gil Joaquina , Martín-Mazuelos Estrella , Rubio Carmen , Sánchez-Sousa Aurora , Serrano Carmen 
Published: March 2005

ABSTRACT

The ability of Sensititre YeastOne to discriminate isolates with reduced caspofungin susceptibility was determined against 36
Candida
spp. (6 with a known
FKS1
mutation). Results were compared with those of M27-A2. The MIC endpoint was 100% growth inhibition. Overall agreement (±2 log
2
) was 87.16%. Sensititre YeastOne detected strains with reduced caspofungin susceptibility.
Genetic Basis of Antibiotic Resistance in Clinical Isolates ofStreptococcus gallolyticus(Streptococcus bovis)
Authors: Leclercq Roland , Huet Corinne , Picherot Mélanie , Trieu-Cuot Patrick , Poyart Claire 
Published: March 2005

ABSTRACTAmong 128Streptococcus gallolyticus(Streptococcus bovis) isolates, 77.7% were resistant to tetracyclines and containedtet(M) and/ortet(L) and/ortet(O). A total of 59.4% had macrolide resistance and containederm(B) and, rarely,mef(A). Among the one-third of isolates highly resistant to kanamycin and streptomycin, most harboredaphA3andaad-6genes.
Effects of Age and Sex on Single-Dose Pharmacokinetics of Tigecycline in Healthy Subjects
Authors: Muralidharan Gopal , Fruncillo Richard J. , Micalizzi Marlynne , Raible Donald G. , Troy Steven M. 
Published: March 2005

ABSTRACT
The pharmacokinetics of tigecycline was evaluated in 46 healthy young and elderly men and women. Except for the volumes of distribution at steady state (approximately 350 liters in women versus 500 liters in men), there were no significant differences in tigecycline pharmacokinetic parameters. Based on pharmacokinetics, no dosage adjustment is warranted based on age or sex.
Chlamydial Infection Induces Pathobiotype-Specific Protein Tyrosine Phosphorylation in Epithelial Cells
Authors: Virok Dezso P. , Nelson David E. , Whitmire William M. , Crane Deborah D. , Goheen Morgan M. , Caldwell Harlan D. 
Published: March 2005

ABSTRACT

Members of the genus
Chlamydia
are strict obligate intracellular pathogens that exhibit marked differences in host range and tissue tropism despite sharing a remarkable level of genomic synteny. These pathobiotype differences among chlamydiae are also mirrored in their early interactions with cultured mammalian host cells. Chlamydial attachment and entry is known to trigger protein tyrosine phosphorylation. In this study, we examined the kinetics and pattern of protein tyrosine phosphorylation induced by infection with a comprehensive collection of chlamydial strains exhibiting diversity in host, tissue, and disease tropisms. We report new findings showing that protein tyrosine phosphorylation patterns induced by infection directly correlate with the pathobiotype of the infecting organism. Patterns of protein tyrosine phosphorylation were induced following early infection that unambiguously categorized chlamydial pathobiotypes into four distinct groups: (i)
Chlamydia trachomatis
trachoma biovars (serovars A to H), (ii)
C. trachomatis
lymphogranuloma venereum biovars (serovars L1 to L3), (iii)
C. muridarum
, and (iv)
C. pneumoniae
and
C. caviae
. Notably, chlamydia-infected murine and human epithelial cells exhibited the same protein tyrosine phosphorylation patterns; this is indirect evidence suggesting that the phosphorylated protein(s) is of chlamydial origin. If our hypothesis is correct, these heretofore-uncharacterized proteins may represent a novel class of bacterial molecules that influence pathogen-host range or tissue tropism.
Antibiotic-Free Plasmid Stabilization by Operator-Repressor Titration for Vaccine Delivery by Using Live
Salmonella enterica
Serovar Typhimurium
Authors: Garmory Helen S. , Leckenby Matthew W. , Griffin Kate F. , Elvin Stephen J. , Taylor Rosa R. , Hartley M. Gill , Hanak Julian A. J. , Williamson E. Diane , Cranenburgh Rocky M. 
Published: March 2005

ABSTRACT

Live, attenuated bacteria are effective vectors for heterologous antigen delivery. However, loss of heterologous gene-bearing plasmids is problematic, and antibiotics and their resistance genes are not desirable for in vivo DNA vaccine delivery due to biosafety and regulatory concerns. To solve this problem, we engineered the first vaccine delivery strain that has no requirement for antibiotics or other selectable marker genes to maintain the recombinant plasmid. This model strain of
Salmonella enterica
serovar Typhimurium, SLDAPD, uses operator-repressor titration (ORT) technology, which requires only the short, nonexpressed
lacO
sequence for selection and maintenance. SLDAPD, recovered from the spleens and Peyer's patches of mice following oral inoculation, was shown to maintain a plasmid that, in contrast, was lost from parental strain SL3261. We also demonstrated successful application of this technology to vaccine development, since SLDAPD carrying a plasmid without an antibiotic resistance gene that expressed the
Yersinia pestis
F1 antigen was as efficacious in protecting vaccinated mice against plague as the parental SL3261 strain carrying an antibiotic-selected version of this plasmid. Protection of mice against plague by immunization with
Salmonella
expressing F1 has previously required two or more doses; here we demonstrated for the first time protective immunity after a single oral immunization. This technology can easily be used to convert any suitable attenuated strain to an antibiotic-free ORT strain for recombinant protein vaccine delivery in humans.
Process Development and Analysis of Liver-Stage Antigen 1, a Preerythrocyte-Stage Protein-Based Vaccine forPlasmodium falciparum
Authors: Hillier Collette J. , Ware Lisa A. , Barbosa Arnoldo , Angov Evelina , Lyon Jeffrey A. , Heppner D. Gray , Lanar David E. 
Published: March 2005

ABSTRACTPlasmodium falciparumliver-stage antigen 1 (LSA-1) is expressed solely in infected hepatocytes and is thought to have a role in liver schizogony and merozoite release. Specific humoral, cellular, and cytokine immune responses to LSA-1 are well documented, with epitopes identified that correlate with antibody production, proliferative T-cell responses, or cytokine induction. With the goal of developing a vaccine against this preerythrocyte-stage protein, we undertook the good manufacturing practices (GMP) manufacture of a recombinant LSA-1 construct, LSA-NRC, incorporating the N- and C-terminal regions of the protein and two of the centrally placed 17-amino-acid repeats. To improve the protein yield, a method of codon harmonization was employed to reengineer the gene sequence for expression inEscherichia coli. A 300-liter GMP fermentation produced 8 kg of bacterial cell paste, and a three-step column chromatographic method yielded 8 mg of purified antigen per g of paste. The final bulk protein was >98% pure, demonstrated long-term stability, and contained <0.005 endotoxin units per 50 μg of protein. To accomplish the initial stages of evaluation of this protein as a human-use vaccine against malaria, we immunized rabbits and mice with LSA-NRC in Montanide ISA 720. New Zealand White rabbits and A/J (H-2K) mice produced high-titer antibodies that recognized liver-stage parasites in infected cultured human hepatocytes. Gamma interferon-producing cells, which have been associated with LSA-1-mediated protection, were detected in splenocytes harvested from immunized mice. Finally, sera taken from people living in a region where malaria is holoendemic recognized LSA-NRC by Western blotting.