bluuebunny/crossref_metadata_2025_split huggingface dataset:
Zoledronate has an antitumor effect and induces actin rearrangement in dexamethasone‐resistant myeloma cells
Authors: Koizumi Masayuki , Nakaseko Chiaki , Ohwada Chikako , Takeuchi Masahiro , Ozawa Shinichi , Shimizu Naomi , Cho Ryuko , Nishimura Miki , Saito Yasushi 
Published: September 2007

AbstractNew strategies are needed to overcome the resistance of multiple myeloma (MM) to dexamethasone (Dex). Several recent in vitro studies demonstrated the antitumor effect of nitrogen‐containing amino‐bisphosphonates (N‐BPs) in various tumor cell lines. Inhibition of the prenylation of small G proteins is assumed to be one of the principal mechanisms by which N‐BPs exert their effects. There have been few reports on N‐BP treatment of MM cells that are resistant to Dex. Additionally, it is not known how small G proteins are altered in N‐BP‐treated MM cells. In this study, we evaluated the effect of the most potent N‐BP, zoledronate (ZOL), on a Dex‐resistant human MM cell subline (Dex‐R) that we established from the well‐documented RPMI8226 cell line. ZOL reduced the viability and induced apoptosis of Dex‐R cells. Some of the ZOL‐treated RPMI8226 cells and ZOL‐treated Dex‐R cells were elongated; however, elongated cells were not seen among the Dex‐treated RPMI8226 cells. Furthermore, we found that portions of the small G proteins, Rho and Rap1A, were unprenylated in the ZOL‐treated MM cells. Geranylgeraniol reduced the above‐mentioned ZOL‐induced effects. These findings suggest that ZOL may be beneficial for the treatment of Dex‐resistant MM by suppressing the processing of RhoA and Rap1A.
The clinical significance of the spectrum of interactions of CAP+1 (A→C), a silent β‐globin gene mutation, with other β‐thalassemia mutations and globin gene modifiers in north Indians
Authors: Garewal Gurjeewan , Das Reena , Awasthi Anshu , Ahluwalia Jasmina , Marwaha Ram Kumar 
Published: September 2007

Abstract Objectives: To assess the clinical significance of the interactions of CAP+1 (A→C), a silent β‐globin gene mutation, with other β‐thalassemia mutations and globin gene modifiers in north Indians. Methods: The clinical phenotypes associated with compound heterozygosity for the CAP+1 (A→C) mutation with other β‐thalassemia mutations, together with the potential effect of the genetic modifiers α‐thalassemia and the Xmn‐1Gγ C→T polymorphism were studied in 30 patients. The frequency of the CAP+1 (A→C) polymorphism was determined and an analysis of the red cell indices, HbA2 levels, iron status, and α‐globin genes was carried out in 35 heterozygotes. Results: Based on an analysis of 1075 β‐thalassemia alleles the CAP+1 (A→C) mutation constituted 3.2% of north Indians. There was a wide spectrum of phenotypic severity in compound heterozygotes; 18 of 30 were transfusion dependent. There was a very high frequency of the −/− genotype of the Xmn‐1Gγ polymorphism in compound heterozygotes. Analysis of 35 heterozygotes indicated that approximately half were hematologically normal and therefore genuine ‘silent’ carriers. Conclusions: Compound heterozygotes for CAP+1 (A→C) and other severe β‐thalassemia alleles are phenotypically severe enough to necessitate appropriate therapy and counseling. The unexpected severity of these interactions may be due, in part, to the high frequency of β‐thalassemia alleles associated with the Xmn‐1Gγ− allele in Indian populations. It is concluded that the CAP+1 (A→C) mutation can pose serious difficulties in screening and counseling programs in populations in which it occurs at a significant frequency.
Partial splenic embolization in children with hereditary spherocytosis
Authors: Pratl Barbara , Benesch Martin , Lackner Herwig , Portugaller Horst Rupert , Pusswald Bernadette , Sovinz Petra , Schwinger Wolfgang , Moser Andrea , Urban Christian 
Published: October 2007

AbstractObjectives:  Although total splenectomy is able to reduce clinical symptoms in patients with hereditary spherocytosis (HS), splenectomized patients are at risk to develop overwhelming bacterial infections and, to a lesser extent, thromboembolic complications. In contrast, partial splenectomy or partial splenic embolization (PSE) may also decrease the rate of hemolytic complications while maintaining residual splenic function. The aim of this study was to investigate the benefit of PSE in children with moderate to severe HS.Patients and methods:  We performed PSE via retrograde transfemoral access in eight children (four female, four male) with moderate to severe HS at a median age of 8 yr. HS‐related complications before PSE included gallstones in six and aplastic crises in four children. One patient was transfusion‐dependent.Results:  No acute side effects were seen during or after PSE. Median hemoglobin increased significantly from levels between 7.5 g/dL and 11.65 g/dL before PSE to levels between 8.4 g/dL and 13.35 g/dL after PSE (P = 0.012). Median splenic sizes before PSE ranged from 9.7 cm/m2 to 19.0 cm/m2 and significantly decreased to values between 4.4 cm/m2 and 15.65 cm/m2 during follow‐up (P = 0.012).Conclusions:  PSE appears to be a safe, effective and feasible treatment option for the management of children with moderate to severe HS.
Discovery of a galactic wind in the central region of M100
Authors: Jiménez-Vicente J. , Castillo-Morales A. , Mediavilla E. , Battaner E. 
Published: October 2007

Abstract
We report the discovery of a galactic wind in the central region of the galaxy M100. This result is based on a careful 2D spectroscopic study performed on observations made with the fibre system INTEGRAL on the William Herschel Telescope. The primary evidence of the wind is the presence of blueshifted interstellar NaD absorption lines. The velocity field of the absorbers show a clear rotation pattern but which is globally blueshifted (∼−115 km s−1) with respect to the systemic velocity of the galaxy. The emission lines also present a blueward component arising from the ionized gas phase of the galactic wind. The velocity field of the ionized gas wind component shows no evidence of rotation but exhibits a pattern that can be interpreted in terms of the projection of an outflowing cone or shell. The wind component has [N ii]/Hα ratios of about 1.8, typical of shock ionization. The ionized component of the wind can be identified with an expanding shell of shocked gas, and the neutral component with disc gas entrained in the wind at the interface of the expanding shell with the galactic interstellar medium. The galactic wind seems to be driven uniquely by the nuclear starburst. Our analysis indicates that a non-negligible fraction of the wind material might escape to the intergalactic medium (IGM). In this case, if the wind detected in M100 were representative of similar phenomena in other galaxies with low to moderate activity, the current estimates of metal and dust content of the IGM might be drastically underestimated.
Development of an Internal Control for Evaluation and Standardization of a Quantitative PCR Assay for Detection of
Helicobacter pylori
in Drinking Water
Authors: Sen Keya , Schable Nancy A. , Lye Dennis J. 
Published: September 2007

ABSTRACT

Due to metabolic and morphological changes that can prevent
Helicobacter pylori
cells in water from growing on conventional media, an
H. pylori
-specific TaqMan quantitative PCR (qPCR) assay was developed that uses a 6-carboxyfluorescein-labeled probe (A. E. McDaniels, L. Wymer, C. Rankin, and R. Haugland, Water Res. 39:4808-4816, 2005). However, proper internal controls are needed to provide an accurate estimate of low numbers of
H. pylori
in drinking water. In this study, the 135-bp amplicon described by McDaniels et al. was modified at the probe binding region, using PCR mutagenesis. The fragment was incorporated into a single-copy plasmid to serve as a PCR-positive control and cloned into
Escherichia coli
to serve as a matrix spike. It was shown to have a detection limit of five copies, using a VIC dye-labeled probe. A DNA extraction kit was optimized that allowed sampling of an entire liter of water. Water samples spiked with the recombinant
E. coli
cells were shown to behave like
H. pylori
cells in the qPCR assay. The recombinant
E. coli
cells were optimized to be used at 10 cells/liter of water, where they were shown not to compete with 5 to 3,000 cells of
H. pylori
in a duplex qPCR assay. Four treated drinking water samples spiked with
H. pylori
(100 cells) demonstrated similar cycle threshold values if the chlorine disinfectant was first neutralized by sodium thiosulfate.
Environmental Monitoring of
Mycobacterium bovis
in Badger Feces and Badger Sett Soil by Real-Time PCR, as Confirmed by Immunofluorescence, Immunocapture, and Cultivation
Authors: Sweeney F. P. , Courtenay O. , Hibberd V. , Hewinson R. G. , Reilly L. A. , Gaze W. H. , Wellington E. M. H. 
Published: September 2007

ABSTRACT

Real-time PCR was used to detect and quantify
Mycobacterium bovis
cells in naturally infected soil and badger feces. Immunomagnetic capture, immunofluorescence, and selective culture confirmed species identification and cell viability. These techniques will prove useful for monitoring
M. bovis
in the environment and for elucidating transmission routes between wildlife and cattle.
Evaluation of the
Staphylococcus aureus
Class C Nonspecific Acid Phosphatase (SapS) as a Reporter for Gene Expression and Protein Secretion in Gram-Negative and Gram-Positive Bacteria
Authors: du Plessis Erika , Theron Jacques , Berger Eldie , Louw Maureen 
Published: September 2007

ABSTRACT

A phosphatase secreted by
Staphylococcus aureus
strain 154 has previously been characterized and classified as a new member of the bacterial class C family of nonspecific acid phosphatases. As the acid phosphatase activity can be easily detected with a cost-effective plate screen assay, quantitatively measured by a simple enzyme assay, and detected by zymography, its potential use as a reporter system was investigated. The
S. aureus
acid phosphatase (
sapS
) gene has been cloned and expressed from its own regulatory sequences in
Escherichia coli
,
Bacillus subtilis
, and
Bacillus halodurans
. Transcriptional and translational fusions of the
sapS
gene with selected heterologous promoters and signal sequences were constructed and expressed in all three of the host strains. From the range of promoters evaluated, the strongest promoter for heterologous protein production in each of the host strains was identified, i.e., the
E. coli lacZ
promoter in
E. coli
, the
B. halodurans
alkaline protease promoter in
B. subtilis
, and the
B. halodurans
σ
D
promoter in
B. halodurans
. This is the first report on the development of a class C acid phosphatase gene as a reporter gene with the advantage of being able to function in both gram-positive and gram-negative host strains.
Prevalence of EnterotoxigenicClostridium perfringensIsolates in Pittsburgh (Pennsylvania) Area Soils and Home Kitchens
Authors: Li Jihong , Sayeed Sameera , McClane Bruce A. 
Published: September 2007

ABSTRACTIn the United States and Europe, food poisoning due toClostridium perfringenstype A is predominantly caused byC. perfringensisolates carrying a chromosomal enterotoxin gene (cpe). Neither the reservoir for these isolates nor the point in the food chain where these bacteria contaminate foods is currently understood. Therefore, the current study investigated whether type A isolates carrying a chromosomalcpegene are present in two potential reservoirs, i.e., soil and home kitchen surfaces. NoC. perfringensisolates were recovered from home kitchen surfaces, but most surveyed soil samples containedC. perfringens. The recovered soil isolates were predominantly type A, but some type C, D, and E soil isolates were also identified. Allcpe-positive isolates recovered from soil were genotyped as type A, with theircpegenes oncpeplasmids rather than the chromosome. However, twocpe-positive soil isolates did not carry a classicalcpeplasmid. Both of those atypicalcpe-positive soil isolates were sporulation capable yet failed to produceC. perfringensenterotoxin, possibly because of differences in their upstream promoter regions. Collectively these results suggest that neither soil nor home kitchen surfaces represent major reservoirs for type A isolates with chromosomalcpethat cause food poisoning, although soil does appear to be a reservoir forcpe-positive isolates causing non-food-borne gastrointestinal diseases.
Design and Performance of a 16S rRNA-Targeted Oligonucleotide Probe for Detection of Members of the Genus
Bdellovibrio
by Fluorescence In Situ Hybridization
Authors: Mahmoud Khaled K. , McNeely Damian , Elwood Chelsea , Koval Susan F. 
Published: September 2007

ABSTRACT

A 16S rRNA-targeted, Cy3-labeled oligonucleotide probe was designed to detect members of the genus
Bdellovibrio
by fluorescence in situ hybridization. Specific hybridization conditions were established; however, the detection of bdellovibrios in environmental samples required enrichment, confirming that
Bdellovibrio
spp. are not present in large numbers in the environment.
Analysis of the Mycosamine Biosynthesis and Attachment Genes in the Nystatin Biosynthetic Gene Cluster of
Streptomyces noursei
ATCC 11455
Authors: Nedal Aina , Sletta Håvard , Brautaset Trygve , Borgos Sven E. F. , Sekurova Olga N. , Ellingsen Trond E. , Zotchev Sergey B. 
Published: September 2007

ABSTRACT

The polyene macrolide antibiotic nystatin produced by
Streptomyces noursei
contains a deoxyaminosugar mycosamine moiety attached to the C-19 carbon of the macrolactone ring through the β-glycosidic bond. The nystatin biosynthetic gene cluster contains three genes,
nysDI, nysDII
, and
nysDIII
, encoding enzymes with presumed roles in mycosamine biosynthesis and attachment as glycosyltransferase, aminotransferase, and GDP-mannose dehydratase, respectively. In the present study, the functions of these three genes were analyzed. The recombinant NysDIII protein was expressed in
Escherichia coli
and purified, and its in vitro GDP-mannose dehydratase activity was demonstrated. The
nysDI
and
nysDII
genes were inactivated individually in
S. noursei
, and analyses of the resulting mutants showed that both genes produced nystatinolide and 10-deoxynystatinolide as major products. Expression of the
nysDI
and
nysDII
genes in
trans
in the respective mutants partially restored nystatin biosynthesis in both cases, supporting the predicted roles of these two genes in mycosamine biosynthesis and attachment. Both antifungal and hemolytic activities of the purified nystatinolides were shown to be strongly reduced compared to those of nystatin, confirming the importance of the mycosamine moiety for the biological activity of nystatin.
Nisin-Controlled Extracellular Production of Interleukin-2 inLactococcus lactisStrains, without the Requirement for a Signal Peptide Sequence
Authors: Fernández Antonio , Rodríguez Juan M. , Bongaerts Roy J. , Gasson Michael J. , Horn Nikki 
Published: September 2007

ABSTRACTSecretion of the cytokine interleukin-2 (IL-2) was investigated inLactococcus lactisusing the secretory machinery of the bacteriocin lactococcin A. Surprisingly, thelcnCDtransport genes were not essential for mouse IL-2 secretion. Furthermore, expression of a mature mouse IL-2 gene resulted in interleukin secretion without the requirement for a leader sequence.
Engineering of Artificial Plant Cytochrome P450 Enzymes for Synthesis of Isoflavones by
Escherichia coli
Authors: Leonard Effendi , Koffas Mattheos A. G. 
Published: September 2007

ABSTRACT

Engineered microbes are becoming increasingly important as recombinant production platforms. However, the nonfunctionality of membrane-bound cytochrome P450 enzymes precludes the use of industrially relevant prokaryotes such as
Escherichia coli
for high-level in vivo synthesis of many functional plant-derived compounds. We describe the design of a series of artificial isoflavone synthases that allowed the robust production of plant estrogen pharmaceuticals by
E. coli
. Through this methodology, a plant P450 construct was assembled to mimic the architecture of a self-sufficient bacterial P450 and contained tailor-made membrane recognition signals. The specific in vivo production catalyzed by one identified chimera was up to 20-fold higher than that achieved by the native enzyme expressed in a eukaryotic host and up to 10-fold higher than production by plants. This novel biological device is a strategy for the utilization of laboratory bacteria to robustly manufacture high-value plant P450 products.
Purification and Characterization of a Three-Component Salicylate 1-Hydroxylase from
Sphingomonas
sp. Strain CHY-1
Authors: Jouanneau Yves , Micoud Julien , Meyer Christine 
Published: September 2007

ABSTRACT

In the bacterial degradation of polycyclic aromatic hydrocarbons (PAHs), salicylate hydroxylases catalyze essential reactions at the junction between the so-called upper and lower catabolic pathways. Unlike the salicylate 1-hydroxylase from pseudomonads, which is a well-characterized flavoprotein, the enzyme found in sphingomonads appears to be a three-component Fe-S protein complex, which so far has not been characterized. Here, the salicylate 1-hydroxylase from
Sphingomonas
sp. strain CHY-1 was purified, and its biochemical and catalytic properties were characterized. The oxygenase component, designated PhnII, exhibited an α
3
β
3
heterohexameric structure and contained one Rieske-type [2Fe-2S] cluster and one mononuclear iron per α subunit. In the presence of purified reductase (PhnA4) and ferredoxin (PhnA3) components, PhnII catalyzed the hydroxylation of salicylate to catechol with a maximal specific activity of 0.89 U/mg and showed an apparent

K
m

for salicylate of 1.1 ± 0.2 μM. The hydroxylase exhibited similar activity levels with methylsalicylates and low activity with salicylate analogues bearing additional hydroxyl or electron-withdrawing substituents. PhnII converted anthranilate to 2-aminophenol and exhibited a relatively low affinity for this substrate (

K
m

, 28 ± 6 μM). 1-Hydroxy-2-naphthoate, which is an intermediate in phenanthrene degradation, was not hydroxylated by PhnII, but it induced a high rate of uncoupled oxidation of NADH. It also exerted strong competitive inhibition of salicylate hydroxylation, with a

K
i

of 0.68 μM. The properties of this three-component hydroxylase are compared with those of analogous bacterial hydroxylases and are discussed in light of our current knowledge of PAH degradation by sphingomonads.
Detection of
Histoplasma
Antigen by a Quantitative Enzyme Immunoassay
Authors: Connolly Patricia A. , Durkin Michelle M. , LeMonte Ann M. , Hackett Emily J. , Wheat L. Joseph 
Published: October 2007

ABSTRACT

The second-generation
Histoplasma
antigen immunoassay is semiquantitative, expressing results as a comparison to a negative control, which requires repeat testing of the prior specimen with the current specimen to accurately determine a change in antigen. Reporting results in this manner often is confusing to the ordering physician and laboratory. Development of a quantitative assay could improve accuracy, reduce interassay variability, and eliminate the need to test the prior sample with the current sample in the same assay. Calibrators with known concentrations of
Histoplasma
antigen were used to quantitate antigen in specimens from patients with histoplasmosis and from controls. Samples from cases of disseminated histoplasmosis or other mycoses and controls were tested to evaluate the performance characteristics of the quantitative assay. Paired specimens were evaluated to determine if quantitation eliminated the need to test the current and prior specimens in the same assay to assess a change in antigen. The sensitivity in samples from patients with AIDS and disseminated histoplasmosis was 100% in urine and 92.3% in serum. Cross-reactions occurred in 70% of other endemic mycoses, but not in aspergillosis. Specificity was 99% in controls with community-acquired pneumonia, medical conditions in which histoplasmosis was excluded, or healthy subjects. A change in antigen level categorized as an increase, no change, or decrease based on antigen units determined in the same assay agreed closely with the category of change in nanograms/milliliter determined from testing current and prior specimens in different assays. Sensitivity, specificity, and interassay precision are excellent in the new third-generation quantitative
Histoplasma
antigen immunoassay.
High Prevalence of the CD14-159CC Genotype in Patients Infected with Severe Acute Respiratory Syndrome-Associated Coronavirus
Authors: Yuan Fang F. , Boehm Ingrid , Chan Paul K. S. , Marks Katherine , Tang Julian W. , Hui David S. C. , Sung Joseph J. Y. , Dyer Wayne B. , Geczy Andrew F. , Sullivan John S. 
Published: October 2007

ABSTRACT

To investigate whether genetic factors of innate immunity might influence susceptibility and/or progression in individuals infected with SARS, we evaluated the CD14 gene polymorphism in 198 Hong Kong blood donors and 152 Hong Kong severe acute respiratory syndrome (SARS) patients who were previously genotyped for FcγRIIA polymorphisms. The prevalence of the CD14-159CC polymorphism was significantly higher in the patients with severe SARS than in the those with mild SARS or controls (31% versus 15% [mild SARS] or 20% [controls]; mild SARS:
P
= 0.029; odds ratio, 2.74; 95% confidence interval, 1.15 to 6.57; controls,
P
= 0.04; odds ratio, 2.41; 95% confidence interval, 1.05 to 5.54), and both CD14-159CC and FcγRIIA-RR131 are risk genotypes for severe SARS-CoV infection.
Immunity to
Neisseria meningitidis
Group B in Adults despite Lack of Serum Bactericidal Antibody
Authors: Welsch Jo Anne , Granoff Dan 
Published: October 2007

ABSTRACT

Serum-complement-mediated bactericidal antibody (SBA) remains the serologic hallmark of protection against meningococcal disease, despite experimental and epidemiologic data that SBA may underestimate immunity. We measured bactericidal activity against three strains of
Neisseria meningitidis
group B in sera from 48 healthy adults and in whole blood from 15 subjects. Blood was anticoagulated with lepirudin, a specific thrombin inhibitor not known to activate complement. Depending on the test strain, protective SBA titers of ≥1:4 were present in only 8 to 15% of the subjects, whereas bactericidal activity was present in 40 to 87% of subjects according to the blood assay. Among SBA-negative subjects, blood from 23 to 42% gave a decrease of ≥2 log
10
CFU/ml after 1 h of incubation, and blood from 36 to 83% gave a decrease of ≥1 log
10
after 2 h. For most blood samples, bactericidal antibodies primarily were directed against noncapsular antigens, since activity was not inhibited by group B polysaccharide. For some SBA-negative subjects, white cells were not needed, since similar respective bactericidal activities were observed in blood and plasma. Bactericidal activity by whole blood of SBA-negative subjects can be rapid (<1 h) and effective (≥2 log
10
) and, among all subjects, was four- to sixfold more prevalent than a positive SBA. Thus, while an SBA titer of ≥1:4 predicts protection against meningococcal disease, a titer of <1:4 is poorly predictive of susceptibility. More sensitive assays than SBA are needed to assess protective meningococcal immunity, or we risk underestimating the extent of immunity in the population and the effectiveness of new meningococcal vaccines.