bluuebunny/crossref_metadata_2025_split huggingface dataset:
Randomized Prospective Controlled Trial of Recombinant Granulocyte Colony-Stimulating Factor as Adjunctive Therapy for Limb-Threatening Diabetic Foot Infection
Authors: de Lalla Fausto , Pellizzer Giampietro , Strazzabosco Marco , Martini Zeno , Du Jardin Giovanni , Lora Luciano , Fabris Paolo , Benedetti Paolo , Erle Giuseppe 
Published: July 2002

ABSTRACT

Adult diabetic patients admitted to our Diabetes Center from September 1996 to January 1998 for severe, limb-threatening foot infection were consecutively enrolled in a prospective, randomized, controlled clinical study aimed at assessing the safety and efficacy of recombinant human granulocyte colony-stimulating factor (G-CSF) (lenograstim) as an adjunctive therapy for the standard treatment of diabetic foot infection. Forty patients, all of whom displayed evidence of osteomyelitis and long-standing ulcer infection, were randomized 1:1 to receive either conventional treatment (i.e., antimicrobial therapy plus local treatment) or conventional therapy plus 263 μg of G-CSF subcutaneously daily for 21 days. The empiric antibiotic treatment (a combination of ciprofloxacin plus clindamycin) was further adjusted, when necessary, according to the results of cultures and sensitivity testing. Microbiologic assessment of foot ulcers was performed by both deep-tissue biopsy and swab cultures, performed at enrollment and on days 7 and 21 thereafter. Patients were monitored for 6 months; the major endpoints (i.e., cure, improvement, failure, and amputation) were blindly assessed at weeks 3 and 9. At enrollment, both patient groups were comparable in terms of both demographic and clinical data. None of the G-CSF-treated patients experienced either local or systemic adverse effects. At the 3- and 9-week assessments, no significant differences between the two groups could be observed concerning the number of patients either cured or improved, the number of patients displaying therapeutic failure, or the species and number of microorganisms previously yielded from cultures at day 7 and day 21. Conversely, among this small series of patients the cumulative number of amputations observed after 9 weeks of treatment appeared to be lower in the G-CSF arm; in fact, only three patients (15%) in this group had required amputation, whereas nine patients (45%) in the other group had required amputation (
P
= 0.038). In conclusion, the administration of G-CSF for 3 weeks as an adjunctive therapy for limb-threatening diabetic foot infection was associated with a lower rate of amputation within 9 weeks after the commencement of standard treatment. Further clinical studies aimed at precisely defining the role of this approach to this serious complication of diabetes mellitus appear to be justified.
Nonenzymatic Chloramphenicol Resistance Mediated by IncC Plasmid R55 Is Encoded by a
floR
Gene Variant
Authors: Cloeckaert Axel , Baucheron Sylvie , Chaslus-Dancla Elisabeth 
Published: July 2002

ABSTRACT

The IncC plasmid R55, initially described in the 1970s and isolated from
Klebsiella pneumoniae,
confers nonenzymatic chloramphenicol resistance. The gene coding for this resistance was cloned and sequenced and shows 95 to 97% nucleotide identity with the recently reported
floR
gene from
Salmonella enterica
serovar Typhimurium DT104 and from
Escherichia coli
animal isolates, respectively, conferring cross-resistance to florfenicol.
Chloromethane Utilization Gene Cluster from
Hyphomicrobium chloromethanicum
Strain CM2
T
and Development of Functional Gene Probes To Detect Halomethane-Degrading Bacteria
Authors: McAnulla Craig , Woodall Claire A. , McDonald Ian R. , Studer Alex , Vuilleumier Stephane , Leisinger Thomas , Murrell J. Colin 
Published: July 2002

ABSTRACT

Hyphomicrobium chloromethanicum
CM2
T
, an aerobic methylotrophic member of the α subclass of the class
proteobacteria
, can grow with chloromethane as the sole carbon and energy source.
H. chloromethanicum
possesses an inducible enzyme system for utilization of chloromethane, in which two polypeptides (67-kDa CmuA and 35-kDa CmuB) are expressed. Previously, four genes,
cmuA
,
cmuB
,
cmuC
, and
purU
, were shown to be essential for growth of
Methylobacterium chloromethanicum
on chloromethane. The
cmuA
and
cmuB
genes were used as probes to identify homologs in
H. chloromethanicum
. A
cmu
gene cluster (9.5 kb) in
H. chloromethanicum
contained 10 open reading frames:
folD
(partial),
pduX
,
orf153
,
orf207
,
orf225
,
cmuB
,
cmuC
,
cmuA
,
fmdB
, and
paaE
(partial). CmuA from
H. chloromethanicum
(67 kDa) showed high identity to CmuA from
M. chloromethanicum
and contains an N-terminal methyltransferase domain and a C-terminal corrinoid-binding domain. CmuB from
H. chloromethanicum
is related to a family of methyl transfer proteins and to the CmuB methyltransferase from
M. chloromethanicum
. CmuC from
H. chloromethanicum
shows identity to CmuC from
M. chloromethanicum
and is a putative methyltransferase.
folD
codes for a methylene-tetrahydrofolate cyclohydrolase, which may be involved in the C
1
transfer pathway for carbon assimilation and CO
2
production, and
paaE
codes for a putative redox active protein. Molecular analyses and some preliminary biochemical data indicated that the chloromethane utilization pathway in
H. chloromethanicum
is similar to the corrinoid-dependent methyl transfer system in
M. chloromethanicum
. PCR primers were developed for successful amplification of
cmuA
genes from newly isolated chloromethane utilizers and enrichment cultures.
Gene Cloning and Functional Characterization by Heterologous Expression of the Fructosyltransferase of
Aspergillus sydowi
IAM 2544
Authors: Heyer Arnd G. , Wendenburg Regina 
Published: July 2002

ABSTRACT

We have purified a fructosyltransferase from conidia of the inulin-producing fungus
Aspergillus sydowi
IAM 2544 and obtained peptide sequences from proteolytic fragments of the protein. With degenerated primers, we amplified a PCR fragment that was used to screen a cDNA library. The fructosyltransferase gene from
Aspergillus sydowi
(EMBL accession no.
AJ289046
) is expressed in conidia, while no expression could be detected in mycelia by Northern blot analysis of mycelial RNA. The gene encodes a protein with a calculated molecular mass of 75 kDa that is different from all fructosyltransferases in the databases. The only homology that could be detected was to the invertase of
Aspergillus niger
(EMBL accession no.
L06844
). The gene was functionally expressed in
Escherichia coli
, yeast, and potato plants. With protein extracts from transgenic bacteria and yeast, fructooligosaccharides could be produced in vitro. In transgenic potato plants, inulin molecules of up to 40 hexose units were synthesized in vivo. While in vitro experiments with protein extracts from conidia of
Aspergillus sydowi
yielded the same pattern of oligosaccharides as extracts from transformed bacteria and yeast, in vivo inulin synthesis with fungal conidia leads to the production of a high-molecular-weight polymer.
Fate and Dissemination of
Bacillus subtilis
Spores in a Murine Model
Authors: Hoa Tran Thu , Duc Le Hong , Isticato Rachele , Baccigalupi Loredana , Ricca Ezio , Van Pham Hung , Cutting Simon M. 
Published: July 2002

ABSTRACT

Bacterial spores are being consumed as probiotics, although little is known about their efficacy or mode of action. As a first step in characterizing spore probiotics, we have studied the persistence and dissemination of
Bacillus subtilis
spores given orally to mice. Our results have shown that spores do not appear to disseminate across the mucosal surfaces. However, we found that the number of spores excreted in the feces of mice was, in some experiments, larger than the original inoculum. This was an intriguing result and might be explained by germination of a proportion of the spore inoculum in the intestinal tract, followed by limited rounds of cell growth and then sporulation again. This result raises the interesting question of whether it is the spore or the germinated spore that contributes to the probiotic effect of bacterial spores.
Genetic Differentiation between Sympatric Populations of
Bacillus cereus
and
Bacillus thuringiensis
Authors: Vilas-Boas Gislayne , Sanchis Vincent , Lereclus Didier , Lemos Manoel Victor F. , Bourguet Denis 
Published: July 2002

ABSTRACT

Little is known about genetic exchanges in natural populations of bacteria of the spore-forming
Bacillus cereus
group, because no population genetics studies have been performed with local sympatric populations. We isolated strains of
Bacillus thuringiensis
and
B. cereus
from small samples of soil collected at the same time from two separate geographical sites, one within the forest and the other at the edge of the forest. A total of 100
B. cereus
and 98
B. thuringiensis
strains were isolated and characterized by electrophoresis to determine allelic composition at nine enzymatic loci. We observed genetic differentiation between populations of
B. cereus
and
B. thuringiensis
. Populations of a given
Bacillus
species—
B. thuringiensis
or
B. cereus
—were genetically more similar to each other than to populations of the other
Bacillus
species. Hemolytic activity provided further evidence of this genetic divergence, which remained evident even if putative clones were removed from the data set. Our results suggest that the rate of gene flow was higher between strains of the same species, but that exchanges between
B. cereus
and
B. thuringiensis
were nonetheless possible. Linkage disequilibrium analysis revealed sufficient recombination for
B. cereus
populations to be considered panmictic units. In
B. thuringiensis,
the balance between clonal proliferation and recombination seemed to depend on location. Overall, our data indicate that it is not important for risk assessment purposes to determine whether
B. cereus
and
B. thuringiensis
belong to a single or two species. Assessment of the biosafety of pest control based on
B. thuringiensis
requires evaluation of the extent of genetic exchange between strains in realistic natural conditions.
trans
-Sialidase Inhibition Assay, a Highly Sensitive and Specific Diagnostic Test for Chagas' Disease
Authors: Buchovsky A. S. , Campetella O. , Russomando G. , Franco L. , Oddone R. , Candia N. , Luquetti A. , Gonzalez Cappa S. M. , Leguizamon M. S. 
Published: July 2002

ABSTRACT

For the diagnosis of Chagas' disease, the
trans
-sialidase inhibition assay was able to resolve the results for samples with borderline results, to detect as positive 60% of samples that were negative by conventional serology, and to discriminate idiopathic from chagasic megaviscera or cardiopathy. No cross-reaction with sera from patients with other diseases was observed.
Generation and Serological Characterization of Murine Monoclonal Antibodies against O Antigens from
Acinetobacter
Reference Strains
Authors: Pantophlet Ralph , Brade Lore , Brade Helmut 
Published: July 2002

ABSTRACT

O-antigen-specific monoclonal antibodies were generated against
Acinetobacter
strains from international type culture collections and characterized by enzyme immunoassay and Western and colony blotting. The antibodies aid in the further completion of an O-serotyping scheme for
Acinetobacter
and, due to their high specificity, are especially useful to all working with these strains.
PrP
Sc
Is Not Detected in Peripheral Blood Leukocytes of Scrapie-Infected Sheep: Determining the Limit of Sensitivity by Immunohistochemistry
Authors: Herrmann Lynn M. , Baszler Timothy V. , Knowles Donald P. , Cheevers William P. 
Published: July 2002

ABSTRACT

Peripheral blood leukocytes (PBLs) from scrapie-infected sheep were evaluated for the presence of PrP
Sc
by using dissociated retropharyngeal lymph node (DRLN) cells and immunohistochemistry (IHC). PrP
Sc
-positive cells were detected in 2.05% ± 0.28% of 3 × 10
6
DRLN cells, but were not detected in 3 × 10
6
PBLs from scrapie-infected sheep. Titration of DRLN cells mixed with PBLs showed that IHC detects a minimum of 0.00205% or 60 PrP
Sc
-positive cells in 3 × 10
6
PBLs.
Active Immunization in the United States: Developments over the Past Decade
Authors: Dennehy Penelope H. 
Published: July 2002

SUMMARYThe Centers for Disease Control and Prevention has identified immunization as the most important public health advance of the 20th century. The purpose of this article is to review the changes that have taken place in active immunization in the United States over the past decade. Since 1990, new vaccines have become available to prevent five infectious diseases: varicella, rotavirus, hepatitis A, Lyme disease, and Japanese encephalitis virus infection. Improved vaccines have been developed to prevent Haemophilus influenzae type b, pneumococcus, pertussis, rabies, and typhoid infections. Immunization strategies for the prevention of hepatitis B, measles, meningococcal infections, and poliomyelitis have changed as a result of the changing epidemiology of these diseases. Combination vaccines are being developed to facilitate the delivery of multiple antigens, and improved vaccines are under development for cholera, influenza, and meningococcal disease. Major advances in molecular biology have enabled scientists to devise new approaches to the development of vaccines against diseases ranging from respiratory viral to enteric bacterial infections that continue to plague the world's population.
Streptococcus suis
Serotype 2 Interactions with Human Brain Microvascular Endothelial Cells
Authors: Charland Nathalie , Nizet Victor , Rubens Craig E. , Kim Kwang Sik , Lacouture Sonia , Gottschalk Marcelo 
Published: July 2002

ABSTRACT

Streptococcus suis
serotype 2 is a worldwide causative agent of many forms of swine infection and is also recognized as a zoonotic agent causing human disease, including meningitis. The pathogenesis of
S. suis
infections is poorly understood. Bacteria circulate in the bloodstream in the nonimmune host until they come in contact with brain microvascular endothelial cells (BMEC) forming the blood-brain barrier. The bacterial polysaccharide capsule confers antiphagocytic properties. It is known that group B streptococci (GBS) invade and damage BMEC, which may be a primary step in the pathogenesis of neonatal meningitis. Interactions between
S. suis
and human endothelial cells were studied to determine if they differ from those between GBS and endothelial cells. Invasion assays performed with BMEC and human umbilical vein endothelial cells demonstrated that unlike GBS,
S. suis
serotype 2 could not invade either type of cell. Adherence assays showed that
S. suis
adhered only to BMEC, whereas GBS adhered to both types of cell. These interactions were not affected by the presence of a capsule, since acapsular mutants from both bacterial species adhered similarly compared to the wild-type strains. Lactate dehydrogenase release measurements indicated that some
S. suis
strains were highly cytotoxic for BMEC, even more than GBS, whereas others were not toxic at all. Cell damage was related to suilysin (
S. suis
hemolysin) production, since only suilysin-producing strains were cytotoxic and cytotoxicity could be inhibited by cholesterol and antisuilysin antibodies. It is possible that hemolysin-positive
S. suis
strains use adherence and suilysin-induced BMEC injury, as opposed to direct cellular invasion, to proceed from the circulation to the central nervous system.
Expression and Immunogenicity of Hemagglutinin A from
Porphyromonas gingivalis
in an Avirulent
Salmonella enterica
Serovar Typhimurium Vaccine Strain
Authors: Kozarov Emil , Miyashita Naohisa , Burks Jacob , Cerveny Karen , Brown Thomas A. , McArthur William P. , Progulske-Fox Ann 
Published: July 2002

ABSTRACT

Porphyromonas gingivalis
is a major etiologic agent of periodontitis, a chronic inflammatory disease that ultimately results in the loss of the supporting tissues of the teeth. Previous work has demonstrated the usefulness of avirulent
Salmonella enterica
serovar Typhimurium strains as antigen delivery systems for protective antigens of pathogens that colonize or cross mucosal surfaces. In this study, we constructed and characterized a recombinant
S. enterica
serovar Typhimurium avirulent vaccine strain which expresses hemagglutinin A and carries no antibiotic resistance markers. HagA, a major virulence-associated surface protein, is a potentially useful immunogen that contains an antigenic epitope which, in humans, elicits an immune response that is protective against subsequent colonization by
P. gingivalis
. The
hagA
gene, including its promoter, was cloned into a balanced-lethal
Salmonella
vector and transferred to the vaccine strain. Heterologous expression of HagA was demonstrated in both
Escherichia coli
JM109 and
S. enterica
serovar Typhimurium vaccine strain χ4072. The HagA epitope was present in its native configuration as determined by immunochemistry and immunoelectron microscopy. Purified recombinant HagA was recognized by sera from mice immunized with the
S. enterica
serovar Typhimurium vaccine strain. The HagA-specific antigen of the vaccine was also found to be recognized by serum from a periodontal patient. This vaccine strain, which expresses the functional hemagglutinin protein, induces a humoral immune response against HagA and may be useful for developing a protective vaccine against periodontal diseases associated with
P. gingivalis
.
Interferon Consensus Sequence Binding Protein Confers Resistance againstYersinia enterocolitica
Authors: Hein Joachim , Kempf Volkhard A. J. , Diebold Joachim , Bücheler Nicole , Preger Sonja , Horak Ivan , Sing Andreas , Kramer Uwe , Autenrieth Ingo B. 
Published: July 2002

ABSTRACTInterferon consensus sequence binding protein (ICSBP)-deficient mice display enhanced susceptibility to intracellular pathogens. At least two distinct immunoregulatory defects are responsible for this phenotype. First, diminished production of reactive oxygen intermediates in macrophages results in impaired intracellular killing of microorganisms. Second, defective early interleukin-12 (IL-12) production upon microbial challenge leads to a failure in gamma interferon (IFN-γ) induction and subsequently in T helper 1 immune responses. Here, we investigated the role of ICSBP in resistance against the extracellular bacteriumYersinia enterocolitica. ICSBP−/−mice failed to produce IL-12 and IFN-γ, but also IL-4, afterYersiniachallenge. In addition, granuloma formation was highly disturbed in infected ICSBP−/−mice, leading to multiple necrotic abscesses in affected organs. Consequently, ICSBP−/−mice rapidly succumbed to acuteYersiniainfection. In vitro treatment of spleen cells from ICSBP−/−mice with recombinant IL-12 (rIL-12) or rIL-18 in combination with a second stimulus resulted in IFN-γ induction. In experimental therapy of infected ICSBP−/−mice, we observed that administration of rIL-12 induced IFN-γ production which was associated with improved resistance toYersinia. In contrast, treatment with rIL-18 failed to enhance endogenous IFN-γ production but nevertheless reduced bacterial burden in ICSBP−/−mice. Although cytokine therapy with rIL-12 or rIL-18 ameliorated the course ofYersiniainfection in ICSBP−/−mice, both cytokines failed to completely restore impaired immunity. Taken together, the results indicate that the transcription factor ICSBP is essential for efficient host immune defense againstYersinia. These results are important for understanding the complex host immune responses in bacterial infections.
Identification of Secreted Proteins ofMycobacterium tuberculosisby a Bioinformatic Approach
Authors: Gomez Manuel , Johnson Sadie , Gennaro Maria Laura 
Published: July 2002

ABSTRACTProteins secreted byMycobacterium tuberculosisare usually targets of immune responses in the infected host. Here we describe a search for secreted proteins that combined the use of bioinformatics andphoA′ fusion technology. The 3,924 proteins deduced from theM. tuberculosisgenome were analyzed with several computer programs. We identified 52 proteins carrying an NH2-terminal secretory signal peptide but lacking additional membrane-anchoring moieties. Of these 52 proteins—the TM1 subgroup—only 7 had been previously reported to be secreted proteins. Our predictions were confirmed in 9 of 10 TM1 genes that were fused toEscherichia coli phoA′, a marker of subcellular localization. These findings demonstrate that the systematic computer search described in this work identified secreted proteins ofM. tuberculosiswith high efficiency and 90% accuracy.
Inactivation of
Pasteurella
(
Mannheimia
)
haemolytica
Leukotoxin Causes Partial Attenuation of Virulence in a Calf Challenge Model
Authors: Highlander Sarah K. , Fedorova Natalie D. , Dusek David M. , Panciera Roger , Alvarez Laura E. , Rinehart Carol 
Published: July 2002

ABSTRACT

The leukotoxin of
Pasteurella
(
Mannheimia
)
haemolytica
is believed to play a significant role in pathogenesis, causing cell lysis and apoptosis that lead to the lung pathology characteristic of bovine shipping fever. Using a system for Cre-
lox
recombination, a nonpolar mutation within the
lktC
transacylase gene of the leukotoxin operon was created. The
lktC
locus was insertionally inactivated using a
loxP-aph3-loxP
cassette, and then the
aph3
marker was excised from the chromosome by Cre recombinase expressed from a
P. haemolytica
plasmid. The resulting
lktC
strain (SH2099) secretes inactive leukotoxin and carries no known antibiotic resistance genes. Strain SH2099 was tested for virulence in a calf challenge model. We inoculated 3 × 10
8
or 3 × 10
9
CFU of wild-type or mutant bacteria into the lungs of healthy, colostrum-deprived calves via transthoracic injection. Animals were observed for clinical signs and for nasal colonization for 4 days, after which they were euthanized and necropsied. The lower inoculum (3 × 10
8
CFU) caused significantly fewer deaths and allowed lung pathology to be scored and compared, while the 3 × 10
9
CFU dose of either the wild-type or mutant was lethal to ≥50% of the calves. The estimated 50% lethal dose of SH2099 was four times higher than that of the wild-type strain. Lung lesion scores were reduced twofold in animals inoculated with the mutant, while clinical scores were nearly equivalent for both strains. The wild-type and mutant strains were equally capable of colonizing the upper respiratory tracts of the calves. In this study, the
P. haemolytica lktC
mutant was shown to be less virulent than the parent strain.