bluuebunny/crossref_metadata_2025 huggingface dataset:
GTPCyclohydrolase I: Purification, Characterization, and Effects ofInhibition on Nitric Oxide Synthase in
Nocardia
Species
Authors: He Aimin , Rosazza John P. N.
Published: December 2003

GTP
cyclohydrolase I (GTPCH) catalyzes the first step in pteridine
biosynthesis in
Nocardia
sp. strain NRRL 5646.
This enzyme is important in the biosynthesis of tetrahydrobiopterin
(BH
4
), a reducing cofactor required for nitric oxide
synthase (NOS) and other enzyme systems in this organism. GTPCH was
purified more than 5,000-fold to apparent homogeneity by a combination
of ammonium sulfate fractionation, GTP-agarose, DEAE Sepharose, and
Ultragel AcA 34 chromatography. The purified enzyme gave a single band
for a protein estimated to be 32 kDa by sodium dodecyl
sulfate-polyacrylamide gel electrophoresis. The molecular mass of the
native enzyme was estimated to be 253 kDa by gel filtration, indicating
that the active enzyme is a homo-octamer. The enzyme follows
Michaelis-Menten kinetics, with a

K
m

for GTP of 6.5μ
M.
Nocardia
GTPCH possessed a unique N-terminal amino
acid sequence. The pH and temperature optima for the enzyme were 7.8
and 56°C, respectively. The enzyme was heat stable and slightly
activated by potassium ion but was inhibited by calcium, copper, zinc,
and mercury, but not magnesium. BH
4
inhibited enzyme
activity by 25% at a concentration of 100 μM.
2,4-Diamino-6-hydroxypyrimidine (DAHP) appeared to competitively
inhibit the enzyme, with a

K
i

of 0.23 mM. With
Nocardia
cultures, DAHP decreased medium levels of
NO
2
−
plus
NO
3
−
. Results suggest that in
Nocardia
cells, NOS synthesis of nitric oxide is indirectly
decreased by reducing the biosynthesis of an essential reducing
cofactor,
BH
4
.
Effect of Food Processing-Related Stresses on AcidTolerance of
Listeriamonocytogenes
Authors: Koutsoumanis Konstantinos P. , Kendall Patricia A. , Sofos John N.
Published: December 2003

Stationary-phase
cells of
Listeria monocytogenes
grown in glucose-free or
glucose-containing media were exposed for 90 min to various stresses,
including acid stress (pH 4.0 to 7.0), osmotic stress (10.5 to
20.5% NaCl), and various temperatures (−5 to
50°C), and were further exposed to pH 3.5. Exposure to a mildly
acidic (pH 5.0 to 6.0) environment provided protection of the pathogen
against acid upon subsequent exposure. This adaptive response, however,
was found to be strongly dependent on other environmental conditions
during the shock, such as temperature or the simultaneous presence of a
second stress factor (NaCl). Growth of
L. monocytogenes
in the
presence of glucose resulted in enhanced survival of the pathogen at pH
3.5. Sublethal stresses other than acidic stresses, i.e., osmotic,
heat, and low-temperature stresses, did not affect the acid resistance
of
L. monocytogenes
(
P
> 0.5). More-severe
levels of these stresses, however, resulted in sensitization of the
pathogen to
acid.
Avian Reservoirs and Zoonotic Potential of theEmerging Human Pathogen
Helicobactercanadensis
Authors: Waldenström Jonas , On Stephen L. W. , Ottvall Richard , Hasselquist Dennis , Harrington Clare S. , Olsen Björn
Published: December 2003

A
polyphasic identification approach was used to investigate the
taxonomic position of
Campylobacter-
like isolates recovered
from barnacle geese (
Branta leucopsis
) and Canada geese
(
Branta candensis
). Seven strains were selected from a
collection of 21 isolates and analyzed by extensive phenotypic testing;
four strains were characterized by 16S rRNA gene sequence analysis. The
results clearly identified the bird isolates as
Helicobacter
canadensis
, recently described as an emerging human pathogen. This
is the first report of an animal reservoir for this organism and of its
presence in Europe and confirms the zoonotic potential of
H.
canadensis
.
Chemoattraction of
Vibrio fischeri
to Serine, Nucleosides, and
N
-Acetylneuraminic Acid, a Component of Squid Light-Organ Mucus
Authors: DeLoney-Marino Cindy R. , Wolfe Alan J. , Visick Karen L.
Published: December 2003

Newlyhatched juveniles of the Hawaiian squid
Euprymna scolopes
rapidly become colonized by the bioluminescent marine bacterium
Vibrio fischeri
. Motility is required to establish the
symbiotic colonization, but the role of chemotaxis is unknown. In this
study we analyzed chemotaxis of
V. fischeri
to a number of
potential attractants. The bacterium migrated toward serine and most
sugars tested.
V. fischeri
also exhibited the unusual ability
to migrate to nucleosides and nucleotides as well as to

N

-acetylneuraminic acid, a component of squid
mucus.
Detection of a
Salmonella enterica
SerovarCalifornia Strain Spreading in Spanish Feed Mills and GeneticCharacterization with DNAMicroarrays
Authors: Alvarez Juan , Porwollik Steffen , Laconcha Idoia , Gisakis Vassilis , Vivanco Ana Belén , Gonzalez Iratxe , Echenagusia Susana , Zabala Nieves , Blackmer Felisa , McClelland Michael , Rementeria Aitor , Garaizar Javier
Published: December 2003

We
performed an epidemiological study on
Salmonella
isolated from
raw plant-based feed in Spanish mills. Overall, 32
different
Salmonella
serovars were detected. Despite its rare
occurrence in humans and animals,
Salmonella enterica
serovar California was found to be the predominant serovar in Spanish
feed mills. Different typing techniques showed that isolates of this
serovar were genetically closely related, and comparative genomic
hybridization using microarray technology revealed 23
S
.
enterica
serovar Typhimurium LT2 gene clusters that are absent
from serovar
California.
Application of Genome-Wide Expression Analysis ToIdentify Molecular Markers Useful in MonitoringIndustrialFermentations
Authors: Higgins Vincent J. , Rogers Peter J. , Dawes Ian W.
Published: December 2003

Genome-wide
expression analysis of an industrial strain of
Saccharomyces
cerevisiae
identified the
YOR387c
and
YGL258w
homologues as highly inducible in zinc-depleted
conditions. Induction was specific for zinc deficiency and was
dependent on Zap1p. The results indicate that these sequences may be
valuable molecular markers for detecting zinc deficiency in industrial
fermentations.
Prevalence of Cholera Toxin Genes (
ctxA
and
zot
) among Non-O1/O139
Vibrio cholerae
Strainsfrom Newport Bay,California
Authors: Jiang Sunny , Chu Weiping , Fu Wuxia
Published: December 2003

The
examination of 137 non-O1/O139
Vibrio cholerae
isolates from
Newport Bay, California, indicated the presence of diverse genotypes
and a temporal succession. Unexpectedly, the cholera toxin gene
(
ctxA
) was found in 17% of the strains, of which
one-third were also positive for the
zot
gene. This suggests
that
ctxA
is prevalent in the region of nonepidemicity and is
likely to have an environmental
origin.
Case Study of the Distribution of Mucosa-Associated
Bifidobacterium
Species,
Lactobacillus
Species, andOther Lactic Acid Bacteria in the HumanColon
Authors: Nielsen D. S. , Møller P. L. , Rosenfeldt V. , Pærregaard A. , Michaelsen K. F. , Jakobsen M.
Published: December 2003

The
distribution of mucosa-associated bacteria, bifidobacteria and
lactobacilli and closely related lactic acid bacteria, in biopsy
samples from the ascending, transverse, and descending parts of the
colon from four individuals was investigated by denaturing gradient gel
electrophoresis (DGGE). Bifidobacterial genus-specific,
Lactobacillus
group-specific, and universal bacterial primers
were used in a nested PCR approach to amplify a fragment of the 16S
rRNA gene. DGGE profiles of the bifidobacterial community were
relatively simple, with one or two amplicons detected at most sampling
sites in the colon. DGGE profiles obtained with
Lactobacillus
group-specific primers were complex and varied with host and sampling
site in the colon. The overall bacterial community varied with host but
not sampling
site.
A Newly DiscoveredVerotoxin Variant, VT2
g
, Produced by Bovine Verocytotoxigenic
Escherichiacoli
Authors: Leung P. H. M. , Peiris J. S. M. , Ng W. W. S. , Robins-Browne R. M. , Bettelheim K. A. , Yam W. C.
Published: December 2003

A
new verotoxin (VT) variant, designated
vt2g
, was identified
from a bovine strain of verocytotoxigenic
Escherichia coli
(VTEC) serotype O2:H25. When
vt2g
was aligned with published
sequences of
vt2
and
vt
variants, it exhibited the
highest DNA sequence homology with
vt2
and
vt2c
.
However,
vt2g
was not detected by
vt2-
specific
primers and probes, although it was partially neutralized by an
antiserum to the VT2A subunit. VT2g was cytotoxic for Vero and HeLa
cells and was not activated by mouse intestinal mucus. The
vt2g
gene was detected in 3 of 409 (0.7%) bovine VTEC
strains, including serotypes O2:H25, O2:H45 and
Ont:H
−
.
Antifungal 3-Hydroxy Fatty Acids from
Lactobacillus plantarum
MiLAB14
Authors: Sjögren Jörgen , Magnusson Jesper , Broberg Anders , Schnürer Johan , Kenne Lennart
Published: December 2003

We
report the identification and chemical characterization of four
antifungal substances, 3-(
R
)-hydroxydecanoic acid,
3-hydroxy-5-
cis
-dodecenoic acid,
3-(
R
)-hydroxydodecanoic acid and
3-(
R
)-hydroxytetradecanoic acid, from
Lactobacillus
plantarum
MiLAB 14. The concentrations of the 3-hydroxy fatty
acids in the supernatant followed the bacterial growth. Racemic
mixtures of the saturated 3-hydroxy fatty acids showed antifungal
activity against different molds and yeasts with MICs between 10 and
100 μg
ml
−1
.
Construction of a Genetically Modified Wine YeastStrain Expressing theAspergillus aculeatus rhaAGene,Encoding an α-l-Rhamnosidase ofEnologicalInterest
Authors: Manzanares Paloma , Orejas Margarita , Gil José Vicente , de Graaff Leo H. , Visser Jaap , Ramón Daniel
Published: December 2003

TheAspergillus aculeatus rhaAgene encoding anα -l-rhamnosidase has been expressed in both laboratory and industrial wine yeast strains. Wines produced in microvinifications, conducted using a combination of the genetically modified industrial strain expressingrhaAand another strain expressing a β-glucosidase, show increased content mainly of the aromatic compound linalool.
Genetic Characterization of the CarotenoidBiosynthetic Pathway in
Methylobacterium extorquens
AM1 andIsolation of a ColorlessMutant
Authors: Van Dien Stephen J. , Marx Christopher J. , O'Brien Brooke N. , Lidstrom Mary E.
Published: December 2003

Genomic
searches were used to reconstruct the putative carotenoid biosynthesis
pathway in the pink-pigmented facultative methylotroph
Methylobacterium extorquens
AM1. Four genes for putative
phytoene desaturases were identified. A colorless mutant was obtained
by transposon mutagenesis, and the insertion was shown to be in one of
the putative phytoene desaturase genes. Mutations in the other three
did not affect color. The tetracycline marker was removed from the
original transposon mutant, resulting in a pigment-free strain with
wild-type growth properties useful as a tool for future
experiments.
Characterization ofSalmonella entericaSerotype Newport Isolated from Humans and Food Animals
Authors: Zhao S. , Qaiyumi S. , Friedman S. , Singh R. , Foley S. L. , White D. G. , McDermott P. F. , Donkar T. , Bolin C. , Munro S. , Baron E. J. , Walker R. D.
Published: December 2003

Salmonella entericaserotype Newport isolates resistant to at least nine antimicrobials (including extended-spectrum cephalosporins), known as serotype Newport MDR-AmpC isolates, have been rapidly emerging as pathogens in both animals and humans throughout the United States. Resistance to extended-spectrum cephalosporins is associated with clinical failures, including death, in patients with systemic infections. In this study, 87Salmonellaserotype Newport strains were characterized by pulsed-field gel electrophoresis (PFGE) and antimicrobial susceptibility testing and examined for the presence of class 1 integrons andblaCMYgenes. Thirty-five PFGE patterns were observed withXbaI, and three of these patterns were indistinguishable among isolates from humans and animals. Fifty-three (60%)Salmonellaserotype Newport isolates were identified as serotype Newport MDR-AmpC, including 16 (53%) of 30 human isolates, 27 (93%) of 29 cattle isolates, 7 (70%) of 10 swine isolates, and 3 (30%) of 10 chicken isolates. However, 28 (32%)Salmonellaserotype Newport isolates were susceptible to all 16 antimicrobials tested. TheblaCMYgene was present in all serotype Newport MDR-AmpC isolates. Furthermore, the plasmid-mediatedblaCMYgene was transferable via conjugation to anEscherichia colistrain. The transconjugant showed the MDR-AmpC resistance profile. Thirty-five (40%) of the isolates possessed class 1 integrons. Sequence analyses of the integrons showed that they containedaadA, which confers resistance to streptomycin, oraadAanddhfr, which confer resistance to trimethoprim-sulfamethoxazole. One integron from a swine isolate contained thesat-1gene, which encodes resistance to streptothricin, an antimicrobial agent that has never been approved for use in the United States. In conclusion,Salmonellaserotype Newport MDR-AmpC was commonly identified amongSalmonellaserotype Newport isolates recovered from humans and food animals. These findings support the possibility of transmission of this organism to humans through the food chain.
Gene Sequence-Based Criteria for Identification of NewRickettsiaIsolates and Description ofRickettsia heilongjiangensissp. nov
Authors: Fournier Pierre-Edouard , Dumler J. Stephen , Greub Gilbert , Zhang Jianzhi , Wu Yimin , Raoult Didier
Published: December 2003

We propose genetic guidelines for the classification of rickettsial isolates at the genus, group, and species levels by using sequences of the 16S rRNA (rrs) gene and four protein-coding genes, thegltA,ompA, andompBgenes and gene D. To be classified as a member of the genusRickettsia, an isolate should exhibit degrees ofrrsandgltAhomology with any of the 20Rickettsiaspecies studied of ≥98.1 and ≥86.5%, respectively. A member of the typhus group should fulfill at least two of the following four criteria: pairwise nucleotide sequence homologies withrrs,gltA,ompB, and gene D of eitherRickettsia typhiorRickettsia prowazekiiof ≥99.4, ≥96.6, ≥92.4, and ≥91.6%, respectively. A member of the spotted fever group should either possess theompAgene or fulfill at least two of the following four criteria: pairwise nucleotide sequence homologies withrrs,gltA,ompB, and gene D of any member of this group of ≥98.8, ≥92.7, ≥85.8, and ≥82.2%, respectively. The existence of a distinct “ancestral” group should be questioned. To be classified as a newRickettsiaspecies, an isolate should not exhibit more than one of the following degrees of nucleotide similarity with the most homologous validated species: ≥99.8 and ≥ 99.9% for therrsandgltAgenes, respectively, and, when amplifiable, ≥98.8, ≥99.2, and ≥99.3% for theompAandompBgenes and gene D, respectively. By use of our classification scheme, “Rickettsia heilongjiangii” belongs to a new species for which we officially propose the nameRickettsia heilongjiangensissp. nov.
Comparison of the Susceptibilities of
Candida
spp. to Fluconazole and Voriconazole in a 4-Year Global Evaluation Using Disk Diffusion
Authors: Hazen Kevin C. , Baron Ellen Jo , Colombo Arnaldo Lopes , Girmenia Corrado , Sanchez-Sousa Aurora , del Palacio Amalia , de Bedout Catalina , Gibbs David L.
Published: December 2003

From June 1997 to December 2001, results of in vitro susceptibility tests of yeast isolates from 35 countries were collected. For 2001 alone, fluconazole results were reported for 22,111 yeast isolates from 77 institutions in 30 countries. Of these isolates, 18,569 were also tested for susceptibility to voriconazole. All study sites tested clinical yeast isolates by recently endorsed NCCLS disk diffusion method M44-P. Disk test plates were automatically read and results were recorded with the BIOMIC Image Analysis System. Species, drug, zone diameter, susceptibility category, MIC, and quality control results were electronically submitted by e-mail quarterly for analysis. Duplicate test results (same patient and same species with same sensitivity-resistance profile and biotype results during any 7-day period) and uncontrolled test results were eliminated from this analysis. The proportion of
Candida albicans
isolates decreased from 69.7% in 1997 to 1998 to 63.0% in 2001, and this decrease was accompanied by a concomitant increase in C.
tropicalis
and
C. parapsilosis
. The susceptibility (susceptible [S]or susceptible-dose dependent [S-DD]) of
C. albicans
isolates to fluconazole was virtually unchanged, from 99.2% in 1997 to 99% in 2001; the
C. glabrata
response to fluconazole was unchanged, from 81.5% S or S-DD in 1997 to 81.7% in 2001, although the percentage of resistant isolates from blood and upper respiratory tract samples appeared to increase over the study period; the percentage of S
C. parapsilosis
isolates decreased slightly, from 98% S or S-DD in 1997 to 96% in 2001; and the percentage of S isolates of
C. tropicalis
increased slightly, from 95.7% in 1997 to 96.9% in 2001. The highest rate of resistance to fluconazole among
C. albicans
isolates was noted in Ecuador (7.6%,
n
= 250). Results from this investigation indicate that the susceptibility of yeast isolates to fluconazole has changed minimally worldwide over the 4.5-year study period and that voriconazole demonstrated 10- to 100-fold greater in vitro activity than fluconazole against most yeast species.
New Methods To Assess Susceptibilities of
Aspergillus
Isolates to Caspofungin
Authors: Imhof Alexander , Balajee S. Arunmozhi , Marr Kieren A.
Published: December 2003

Echinocandins are a group of antifungal agents that target 1,3-β-glucan synthase, causing disruption of mold growth at cells compromising tips and branch points. In part because echinocandins do not induce clear growth inhibition end points using broth dilution techniques, methods to test susceptibility have not yet been standardized. We developed a novel susceptibility assay that measures growth of
Aspergillus
species on solid agar media that contain serial dilutions of caspofungin (agar dilution). Results of agar dilution testing of multiple isolates were compared to results obtained by broth microdilution (MIC), microscopic evaluation (minimal effective concentration [MEC]), and a new method to measure fungal burden, quantification of secreted hyphal antigen. MICs obtained by the agar dilution method were within 1 dilution of MECs for 85% of the
Aspergillus
isolates; the highest agreement was observed for isolates of
Aspergillus niger
(95%), which were particularly susceptible to caspofungin. Agar dilution MICs were also consistent with those obtained by quantifying antigen secretion. MICs obtained by broth microdilution were different than MICs by any other method. Several
Aspergillus
isolates with decreased susceptibility to caspofungin were identified. Agar dilution is simple and reproducible, and results were consistent with the results of more technically demanding techniques. This method may be appropriate for use in the clinical laboratory.
Highly Resistant Metabolically Deficient Dwarf Mutant of
Escherichia coli
Is the Cause of a Chronic Urinary Tract Infection
Authors: Trülzsch Konrad , Hoffmann Harald , Keller Christiane , Schubert Sören , Bader Lutz , Heesemann Jürgen , Roggenkamp Andreas
Published: December 2003

We present the case of a 68-year-old diabetic woman who has been suffering from chronic urinary tract infections, recurring over a period of at least 5 years, caused by a slowly growing metabolically deficient dwarf mutant (MDD) of
Escherichia coli.
This MDD strain was auxotrophic for histidine, was resistant to multiple antibiotics, and showed atypical growth behavior. Colonies were tiny on routine media but were able to revert to normal growth after extended incubation. This strain was identified as
E. coli
by 16S ribosomal DNA sequencing, and virulence factor profiles were determined by PCR. Seven MDD isolates collected over the 5-year period were grown from midstream urine to significant colony counts and shown to belong to the same clonal group by pulsed-field gel electrophoresis and enterobacterial repetitive intergenic consensus PCR. These MDDs were repeatedly misidentified by biochemical methods due to their slow growth and atypical colony morphology. This case highlights the importance of recognizing MDDs of
Enterobacteriaceae
in patients with chronic infections. To our knowledge this is the first report of an MDD of
E. coli
causing a chronic urinary tract infection.
Determinants of Acquisition and Carriage ofStaphylococcus aureusin Infancy
Authors: Peacock Sharon J. , Justice Anita , Griffiths D. , de Silva G. D. I. , Kantzanou M. N. , Crook Derrick , Sleeman Karen , Day Nicholas P. J.
Published: December 2003

Nasal carriage ofStaphylococcus aureusis a major risk factor for invasiveS. aureusdisease. The aim of this study was to define factors associated with carriage. We conducted a prospective, longitudinal community-based study of infants and their mothers for a period of 6 months following delivery. The epidemiology of carriage was examined for 100 infant-mother pairs. Infant carriage varied significantly with age, falling from 40 to 50% during the first 8 weeks to 21% by 6 months. Determinants of infantS. aureuscarriage included maternal carriage, breastfeeding, and number of siblings. Bacterial typing ofS. aureuswas performed by pulsed-field gel electrophoresis and multilocus sequence typing. The majority of individuals carried a single strain ofS. aureusover time, and the mother was the usual source for colonizing isolates in infants. The effect of other components of the normal nasal flora on the development ofS. aureuscarriage was examined in 157 consecutive infants. Negative associations (putative bacterial interference) betweenS. aureusand other species occurred early in infancy but were not sustained. An increasing antistaphylococcal effect observed over time was not attributable to bacterial interference.S. aureuscarriage in infants is likely to be determined by a combination of host, environmental, and bacterial factors, but bacterial interference does not appear to be an ultimate determinant of carrier status.
Molecular Comparison of Isolates of
Burkholderia multivorans
from Patients with Cystic Fibrosis in the United Kingdom
Authors: Turton Jane F. , Kaufmann Mary E. , Mustafa Nazim , Kawa Sonia , Clode Fiona E. , Pitt Tyrone L.
Published: December 2003

Burkholderia multivorans
strains from 47 cystic fibrosis (CF) patients in 28 hospitals were compared by pulsed-field gel electrophoresis (PFGE) and flagellin (
fliC
) PCR-restriction fragment length polymorphism (PCR-RFLP) analysis. A considerable degree of genetic variation was evident, with each patient harboring a strain with a unique PFGE profile. Four sizes of
fliC
amplicons were produced, and these amplicons gave 13 RFLP types with restriction enzyme
Msp
I.
B. multivorans
did not appear to spread between patients, suggesting that most CF patients acquire the organism from the natural environment.
Evaluationof the Mandibular Third Molar Pericoronitis Flora and ItsSusceptibility to Different Antibiotics PrescribedinFrance
Authors: Sixou Jean-Louis , Magaud Christophe , Jolivet-Gougeon Anne , Cormier Michel , Bonnaure-Mallet Martine
Published: December 2003

This
work assessed the polymicrobial flora of mandibular third molar
pericoronitis. Obligate anaerobes were found in almost all cases (32 of
35). Amoxicillin and pristinamycin were the most effective against the
flora, particularly aerobic organisms. Metronidazole alone or combined
with spiramycin was the most effective drug against obligate
anaerobes.
Expression of Equi Merozoite Antigen 2 during Development of
Babesia equi
in the Midgut and Salivary Gland of the Vector Tick
Boophilus microplus
Authors: Ueti Massaro W. , Palmer Guy H. , Kappmeyer Lowell S. , Scoles Glen A. , Knowles Donald P.
Published: December 2003

Equi merozoite antigens 1 and 2 (EMA-1 and EMA-2) are
Babesia equi
proteins expressed on the parasite surface during infection in horses and are orthologues of proteins in
Theileria
spp., which are also tick-transmitted protozoal pathogens. We determined in this study whether EMA-1 and EMA-2 were expressed within the vector tick
Boophilus microplus. B. equi
transitions through multiple, morphologically distinct stages, including sexual stages, and these transitions culminate in the formation of infectious sporozoites in the tick salivary gland. EMA-2-positive
B. equi
stages in the midgut lumen and midgut epithelial cells of
Boophilus microplus
nymphs were identified by reactivity with monoclonal antibody 36/253.21. This monoclonal antibody also recognized
B. equi
in salivary glands of adult
Boophilus microplus
. In addition, quantification of
B. equi
in the mammalian host and vector tick indicated that the duration of tick feeding and parasitemia levels affected the percentage of nymphs that contained morphologically distinct
B. equi
organisms in the midgut. In contrast, there was no conclusive evidence that
B. equi
EMA-1 was expressed in either the
Boophilus microplus
midgut or salivary gland when monoclonal antibody 36/18.57 was used. The expression of
B. equi
EMA-2 in
Boophilus microplus
provides a marker for detecting the various development stages and facilitates the identification of novel stage-specific
Babesia
proteins for testing transmission-blocking immunity.
Invasive Fungal Sinusitis Caused byScytalidium dimidiatumin a Lung Transplant Recipient
Authors: Dunn James J. , Wolfe Michael J. , Trachtenberg Joel , Kriesel John D. , Orlandi Richard R. , Carroll Karen C.
Published: December 2003

We describe a case of invasive fungal sinusitis caused byScytalidium dimidiatumin a lung transplant recipient. Treatment was complicated by renal failure with amphotericin B therapies. Following 6 months of voriconazole treatment, the patient remained radiographically and clinically stable for a short time before dying of respiratory failure precipitated by graft rejection.
Salmonella enterica
Pneumonia in a Patient with Lung Cancer
Authors: Samonis George , Maraki Sofia , Kouroussis Charalambos , Mavroudis Dimitrios , Georgoulias Vassilios
Published: December 2003

A case of life-threatening
Salmonella enterica
serotype Enteritidis pneumonia in a febrile patient with lung cancer is described. The organism was isolated from the sputum, the protected specimen brush material of bronchial secretions, and the stool. Despite the early administration of appropriate and adequate treatment, the patient died 7 days after the onset of the infection.
Should patient consent be required to write a do not resuscitate order?
Authors: Biegler P
Published: December 2003

Consent ought to be required to withhold treatment that is in a patient’s best interests to receive. Do not resuscitate (DNR) orders are examples of best interests assessments at the end of life. Such assessments represent value judgments that cannot be validly ascertained without patient input. If patient input results in that patient dissenting to the DNR order then individual physicians are not justified in overriding such dissent. To do so would give unjustifiable primacy to the values of the individual physician. Therefore patient consent is effectively required to write a DNR order. Patient dissent to a DNR order should trigger a fair process mechanism to resolve the dispute.
Beware the rooster: smokeless tobacco companies who claim they want to help
Authors: Myers M
Published: December 2003

It should not be assumed that smokeless tobacco manufacturers share our public health goals
A strategy for controlling the marketing of tobacco products: a regulated market model
Authors: Borland R
Published: December 2003

Objective:To outline a novel strategy for controlling the tobacco market.

Arguments:More comprehensive controls over the tobacco market are essential and long overdue. Effective controls need to encourage the development of less harmful products; control commercial communication to ensure that potential harms are highlighted relative to any benefits; and provide mechanisms to move consumers away from tobacco use, or at least towards less harmful alternatives. Achieving this by regulating the existing industry is one strategy. This paper puts the case for an alternative: to have marketing controlled by an agency (called here the Tobacco Products Agency, or TPA) which tendered to manufacturers for product and which distributed to retailers in ways that reduce incentives to bend or break the law. The TPA would be backed by legislation that made tobacco a controlled substance with possession sale and use only allowed as permitted by the regulations, which in reality would be only as provided by the TPA.

Conclusions:The overall effect of such a model, which we call a “regulated market model”, would be to eliminate most of the incentives and remaining opportunities for commercial promotion of tobacco and to create incentives to encourage the development of less harmful tobacco products. Such a model preserves the competition inherent in a free market, but directs it towards the challenge of reducing the harm from tobacco use.
The US tobacco control community’s view of the future of tobacco harm reduction
Authors: Warner K E , Martin E G
Published: December 2003

Objective:Tobacco harm reduction (THR) has garnered recent attention due to the introduction of novel nicotine delivery products ostensibly intended to reduce risk for inveterate cigarette smokers. This study evaluates the grassroots tobacco control community’s knowledge, opinions, and beliefs about THR.

Design:A web/mail survey conducted in October and November 2002, with a telephone survey of a sample of non-respondents.

Subjects:The 2833 US based registrants for the 2001 National Conference on Tobacco or Health.

Main outcome measures:Respondents’ awareness of THR, perception of its importance, support for regulation, and perception of which THR products should be recommended to inveterate cigarette smokers.

Results:70% of respondents were aware of THR but respondents had low recognition of specific products at the forefront of the debate, such as Swedish snuff. Half believe THR will reduce smoking cessation and cause nicotine experimentation by children; 63% anticipate unintended adverse side effects. More expect THR to have a negative than a positive impact on health. Large majorities support government regulation of THR and conventional tobacco products, but fewer than 30% expect legislation regulating either. Most would recommend nicotine patches (76%) and gum (70%) to inveterate smokers, but no other product was supported by a majority. Scientists are more supportive of THR than activists, while respondents focusing on national/international issues are more supportive than those concentrating on local/state issues.

Conclusions:Many members of the US tobacco control community are unaware of the THR “movement”, while others possess only rudimentary familiarity with it. If and as THR achieves an increasingly prominent role on the tobacco-or-health scene, this community will have to become educated about THR, and be prepared to advocate for regulatory policies that will maximise the potential for positive outcomes. The potential for negative outcomes remains significant.
Impact of the graphic Canadian warning labels on adult smoking behaviour
Authors: Hammond D , Fong G T , McDonald P W , Cameron R , Brown K S
Published: December 2003

Objective:To assess the impact of graphic Canadian cigarette warning labels on current adult smokers.

Design:A random-digit-dial telephone survey was conducted with 616 adult smokers in south western Ontario, Canada in October/November 2001, with three month follow up.

Main outcome measures:Smoking behaviour (quitting, quit attempts, and reduced smoking), intentions to quit, and salience of the warning labels.

Results:Virtually all smokers (91%) reported having read the warning labels and smokers demonstrated a thorough knowledge of their content. A strong positive relation was observed between a measure of cognitive processing—the extent to which smokers reported reading, thinking about, and discussing the new labels—and smokers’ intentions to quit (odds ratio (OR) 1.11, 95% confidence interval (CI) 1.07 to 1.16; p < 0.001). Most important, cognitive processing predicted cessation behaviour at follow up. Smokers who had read, thought about, and discussed the new labels at baseline were more likely to have quit, made a quit attempt, or reduced their smoking three months later, after adjusting for intentions to quit and smoking status at baseline (OR 1.07, 95% CI 1.03 to 1.12; p < 0.001).

Conclusions:Graphic cigarette warning labels serve as an effective population based smoking cessation intervention. The findings add to the growing literature on health warnings and provide strong support for the effectiveness of Canada’s tobacco labelling policy.
What the public thinks about the tobacco industry and its products
Authors: Ashley M J , Cohen J E
Published: December 2003

Objectives:To assess public attitudes toward the tobacco industry and its products, and to identify predictors of attitudes supportive of tobacco industry denormalisation.

Design:Population based, cross sectional survey.

Setting:Ontario, Canada.

Subjects:Adult population (n = 1607).

Main outcome measures:Eight different facets of tobacco industry denormalisation were assessed. A denormalisation scale was developed to examine predictors of attitudes supportive of tobacco industry denormalisation, using bivariate and multivariate analyses.

Results:Attitudes to the eight facets of tobacco industry denormalisation varied widely. More than half of the respondents supported regulating tobacco as a hazardous product, fining the tobacco industry for earnings from underage smoking, and suing tobacco companies for health care costs caused by tobacco. Majorities also thought that the tobacco industry is dishonest and that cigarettes are too dangerous to be sold at all. Fewer than half of the respondents thought that the tobacco industry is mostly or completely responsible for the health problems smokers have because of smoking and that tobacco companies should be sued for taxes lost from smuggling. In particular, less than a quarter thought that the tobacco industry is most responsible for young people starting to smoke. Non-smoking, knowledge about health effects caused by tobacco, and support for the role of government in health promotion were independent predictors of support for tobacco industry denormalisation.

Conclusions:Although Ontarians are ambivalent toward tobacco industry denormalisation, they are supportive of some measures. Mass media programmes aimed at increasing support for tobacco industry denormalisation and continued monitoring of public attitudes toward this strategy are needed.
A comparison of US and Norwegian regulation of coumarin in tobacco products
Authors: Givel M
Published: December 2003

Objective:This paper examines policy processes regarding why the USA and Norway have not regulated coumarin in tobacco.

Design:A qualitative analysis of all tobacco industry documents regarding coumarin since the 1950s from the 1998 US Master Settlement Agreement and subsequent legal settlements. Additional data were collected from newspaper reports, general internet search engines, journal articles, scholarly reports, court cases, statutes, regulations, and informal correspondence with tobacco control experts in Norway.

Main outcome measure:An overview, summary, and analysis of all documents related to coumarin.

Results:In the USA from 1954 until 1985 when coumarin was reportedly removed from domestic cigarettes, but not from pipe tobacco until 1996, and not at all from imported Indian bidi cigarettes, regulatory efforts were stymied. In Norway, from 1973 to the present, the tobacco industry has never disclosed whether its tobacco products contain coumarin. In both the USA and Norway, the extreme delay and lack of vigorous evidence gathering and significant remedies were caused by tobacco industry assertions that revealing tobacco additives was a violation of trade secrets, and by weak regulatory authority and efforts to regulate coumarin.

Conclusion:Vigorous and expeditious regulatory investigations and remedies for harmful additives in tobacco, such as coumarin, can protect the public health. Astute insider and outsider political advocacy by health advocates is required to hold elected officials and civil servants publicly accountable for failing to enact disclosure laws and to engage in effective regulatory efforts.
Hand rolling cigarette papers as the reference point for regulating cigarette fire safety
Authors: Laugesen M , Duncanson M , Fraser T , McClellan V , Linehan B , Shirley R
Published: December 2003

Aim:To compare the burning characteristics of the tobacco and paper of manufactured and hand rolled cigarettes, and set a fire safety standard of manufacture to largely reduce the fire risk from discarded cigarettes.

Methods:(1) Cigarette extinction test of ignition strength: 40 cigarettes per brand, lit and placed on 15 layers of filter paper, in accordance with ASTM test standard E2187-02. (2) Citrate extracted by 0.1N hydrochloric acid from cigarette papers and from tobacco in manufactured cigarettes, the supernatant analysed by high performance liquid chromatography using ultraviolet visual light spectrophotometer. (3) Survey of 750 nationally representative adults age 18 years and over, by telephone, including 184 smokers.

Materials:(a) New Zealand made Holiday, and Horizon, and US made Marlboro manufactured cigarettes; (b) US manufactured Merit with banded paper; (c) Holiday, Horizon and Marlboro hand rolling tobaccos, hand rolled in Rizla cigarette papers; (d) manufactured cigarettes as in (a), reconstructed using Rizla hand rolling cigarette papers.

Results:1. (a) For each brand of manufactured cigarettes, 40/40 burnt full length; (b) for Merit banded paper cigarettes 29/40 (73%) burnt full length; (c) for each brand of hand rolled cigarettes 0/40 burnt full length; (d) 0/40 manufactured cigarettes reconstructed with Rizla hand rolling paper burnt full length. 2. Citrate content: (a) In manufactured cigarette papers: 0.3–0.8 mg; in tobacco of manufactured cigarettes: Holiday 0, Horizon 0, Marlboro 8.8 mg; (b) Merit: in banded paper 0.418 mg; in tobacco 10.23 mg; (c) In hand rolled cigarettes: in the papers < 0.08 mg; in hand rolled tobacco 13.3–15.0 mg; (d) In hand rolling papers of reconstructed cigarettes: < 0.018 mg. 3. Requiring manufactured cigarettes to compulsorily self-extinguish when left unattended was supported by 67% of smokers, 61% of manufactured cigarette smokers, 82% of hand rolled smokers, and by 68% of non-smokers.

Conclusion:The wrapping paper is a key determinant of whether or not unpuffed cigarettes burn their full length. Using international test methods, popular brands of manufactured cigarettes all burnt full length, but none did so when re-wrapped in hand rolling cigarette paper. This provides a ready-to-hand smoker acceptable standard for reducing ignition potential from manufactured cigarettes, as a basis for regulation or litigation.
Tobacco industry strategy to undermine tobacco control in Finland
Authors: Hiilamo H
Published: December 2003

Objective:To identify and explain tobacco industry strategy in undermining tobacco control measures in Finland and results of these interferences in tobacco policy development during the 1980s and early 1990s.

Methods:Tobacco industry documents, which have been publicly available on the internet as a result of litigation in the USA, were analysed. Documents were sought by Finland and by names of organisations and tobacco control activists. Documents were accessed and assessed between September 2000 and November 2002. Tactics of the tobacco industry activities were categorised as presented by Saloojee and Dagli.

Results:The international tobacco companies utilised similar strategies in Finland as in other industrial markets to fight tobacco control and legislation, the health advocacy movement, and litigation. These activities slowed down the development and implementation of the Tobacco Act in Finland. However, despite the extensive pressure, the industry was not able to prevent the most progressive tobacco legislation in Europe from being passed and coming into force in Finland in 1977 and in 1995.

Conclusion:Denying the health hazards caused by tobacco—despite indisputable scientific evidence—decreased the credibility of the tobacco industry. Strategy of denial was falsely chosen, as health advocacy groups were active both in society and the parliamentary system. The strong influence of the tobacco industry may have in fact increased the visibility of tobacco control in Finland as the litigation process was also drawing attention to negative health effects of tobacco. Therefore the tobacco industry did not manage to convince public opinion. However, the tobacco industry did obtain experience in Finland in how to object to tobacco control measures.
Application of toxicological risk assessment principles to the chemical constituents of cigarette smoke
Authors: Fowles J , Dybing E
Published: December 2003

Objective:To provide a hazard prioritisation for reported chemical constituents of cigarette smoke using toxicological risk assessment principles and assumptions. The purpose is to inform prevention efforts using harm reduction.

Data sources:International Agency for Research on Cancer Monographs; California and US Environmental Protection Agency cancer potency factors (CPFs) and reference exposure levels; scientific journals and government reports from the USA, Canada, and New Zealand.

Study selection:This was an inclusive review of studies reporting yields of cigarette smoke constituents using standard ISO methods.

Data extraction:Where possible, the midpoint of reported ranges of yields was used.

Data synthesis:Data on 158 compounds in cigarette smoke were found. Of these, 45 were known or suspected human carcinogens. Cancer potency factors were available for 40 of these compounds and reference exposure levels (RELs) for non-cancer effects were found for 17. A cancer risk index (CRI) was calculated by multiplying yield levels with CPFs. A non-cancer risk index (NCRI) was calculated by dividing yield levels with RELs. Gas phase constituents dominate both CRI and NCRI for cigarette smoke. The contribution of 1,3-butadiene (BDE) to CRI was more than twice that of the next highest contributing carcinogen (acrylonitrile) using potencies from the State of California EPA. Using those potencies from the USEPA, BDE ranked third behind arsenic and acetaldehyde. A comparison of CRI estimates with estimates of smoking related cancer deaths in the USA showed that the CRI underestimates the observed cancer rates by about fivefold using ISO yields in the exposure estimate.

Conclusions:The application of toxicological risk assessment methods to cigarette smoke provides a plausible and objective framework for the prioritisation of carcinogens and other toxicant hazards in cigarette smoke. However, this framework does not enable the prediction of actual cancer risk for a number of reasons that are discussed. Further, the lack of toxicology data on cardiovascular end points for specific chemicals makes the use of this framework less useful for cardiovascular toxicity. The bases for these priorities need to be constantly re-evaluated as new toxicology information emerges.
Bhutan: the world’s most advanced tobacco control nation?
Authors: Ugen S
Published: December 2003

Significant achievements in the area of tobacco control have been made in the kingdom of Bhutan in the Eastern Himalayas, following the initiation of several tobacco control activities
Assessment of the transferability of biological habitat models for Atlantic salmon parr (Salmo salar)
Authors: Guay J C , Boisclair D , Leclerc M , Lapointe M
Published: December 2003

We assessed the transferability of the habitat suitability index (HSI) and the habitat probabilistic index (HPI) between two rivers. Transferability was measured by the ability of HSI and HPI models developed in the Sainte-Marguerite River to predict the distribution of Atlantic salmon parr (Salmo salar) in the Escoumins River. HSI and HPI were based on the pattern of utilization by fish of water depth, current velocity, and substrate size. HSI was developed using the preference curve approach, and HPI was developed using a multiple logistic regression. Predicted values of HSI and HPI in Escoumins River ranged from 0 (poor habitat) to 1 (excellent habitat). Fish density in habitat patches assigned different HSI or HPI values ranged from 0 to 1 fish·100 m–2. Only HPI adequately predicted local variations in parr density (r2 = 0.84) in habitat patches of Escoumins River. Our results suggest that HSI is less transferable between rivers than HPI. Differences in substrate size between the two rivers is suspected to impede the transferability of the HSI model. We also argue that the mathematical structure of HPI provides a larger degree of flexibility that facilitates its transferability and its potential generalization.
Structural organization of the barley D-hordein locus in comparison with its orthologous regions of wheat genomes
Authors: Gu Yong Qiang , Anderson Olin D , Londeorë Cynthia F , Kong Xiuying , Chibbar Ravindra N , Lazo Gerard R
Published: December 2003

D hordein, a prolamin storage protein of barley endosperms, is highly homologous to the high molecular weight (HWM) glutenin subunits, which are the major determinants of bread-making quality in wheat flour. In hexaploid wheat (AABBDD), each genome contains two paralogous copies of HMW-glutenin genes that encode the x- and y-type HMW-glutenin subunits. Previously, we reported the sequence analysis of a 102-kb genomic region that contains the HMW-glutenin locus of the D genome from Aegilops tauschii, the donor of the D genome of hexaploid wheat. Here, we present the sequence analysis of a 120-kb D-hordein region of the barley genome, a more distantly related member of the Triticeae grass tribe. Comparative sequence analysis revealed that gene content and order are generally conserved. Genes included in both of these orthologous regions are arranged in the following order: a Xa21-like receptor kinase, an endosperm globulin, an HMW prolamin, and a serine (threonine) protein kinase. However, in the wheat D genome, a region containing both the globulin and HMW-glutenin gene was duplicated, indicating that this duplication event occurred after the separation of the wheat and barley genomes. The intergenic regions are divergent with regard to the sequence and structural organization. It was found that different types of retroelements are responsible for the intergenic structure divergence in the wheat and barley genomes. In the barley region, we identified 16 long terminal repeat (LTR) retrotransposons in three distinct nested clusters. These retroelements account for 63% of the contig sequence. In addition, barley D hordein was compared with wheat HMW glutenins in terms of cysteine residue conservation and repeat domain organization.Key words: HMW glutenin, evolution, retrotransposon, comparative genomics.