bluuebunny/crossref_metadata_2025_split huggingface dataset:
Demographic Studies from a National Gaucher Disease Screening Program
Authors: Gagnon Deborah M. , Pergament Eugene , Fine Beth A. 
Published: April 2003

AbstractIn May 1993, the National Gaucher Foundation initiated a nationwide Gaucher disease screening program in an attempt to promote recognition of the disease and to detect previously undiagnosed individuals. The program was based on self‐selection by clinical symptoms of individuals who wanted to be tested for Gaucher disease. Information about symptoms, age, ancestry, gender, and family history was obtained via a self‐report screening form completed by 700 individuals. Individuals designated at “high risk” for Gaucher disease were offered beta‐glucocerebrosidase enzyme assay testing. Twenty‐four of the respondents (3.4%) had Gaucher disease. The most commonly reported symptoms were fatigue (79.4%), bone pain (73.7%), and tendency to bruise (67.8%). The symptoms which showed a statistical difference between the “high risk” and “low risk” groups were liver enlargement (p < 0.005) and unexplained bone fractures (p < 0.03). The mean number of positive symptoms also showed a statistical difference between the groups (4.38 symptoms vs. 3.86 symptoms; p < 0.02). Due to the small sample size, no statistical comparisons were made on the symptomatology of affected vs. unaffected persons, but a descriptive analysis of these two groups is reported.
Client Advocacy and Collaboration with Insurance and Laboratory Testing Agencies: A Case Study Illustrating Genetic Counselor Roles
Authors: Lea Dale Halsey 
Published: April 2003

AbstractGenetic counseling and the many roles a genetic counselor must assume to provide quality genetic health care are becoming increasingly complex, especially when the genetic condition is rare, and DNA testing is costly and not yet routinely used. A case study of a couple with a family history of retinoblastoma and their pursuit of DNA testing for prenatal diagnosis is presented. The case study illustrates the instrumental role of the genetic counselor in advocating for clients for genetic services such as DNA testing and in educating insurance companies in the nature and importance of such services to achieve improved client health care outcomes.
Molecular Epidemiology ofMycoplasma conjunctivaein Caprinae: Transmission across Species in Natural Outbreaks
Authors: Belloy Luc , Janovsky Martin , Vilei Edy M. , Pilo Paola , Giacometti Marco , Frey Joachim 
Published: April 2003

ABSTRACTMycoplasma conjunctivaeis the etiological agent of infectious keratoconjunctivitis, a highly contagious ocular infection that affects both domestic and wild Caprinae species in the European Alps. In order to study the transmission and spread ofM. conjunctivaeacross domestic and wild Caprinae populations, we developed a molecular method for subtyping and identifying strains ofM. conjunctivae. This method is based on DNA sequence determination of a variable domain within the genelppS, a gene that encodes an antigenic lipoprotein ofM. conjunctivae. This domain oflppSshows variations among different strains but remains constant upon generations of individual strains on growth medium and thus allows identification of individual strains and estimation of their phylogenetic intercorrelations. The variable domain oflppSis amplified by PCR using primers that match conserved sequences oflppSflanking it. Sequence analysis of the amplified fragment enables fine subtyping ofM. conjunctivaestrains. The method is applicable both to isolated strains and to clinical samples directly without requiring the cultivation of the strain. Using this method, we show thatM. conjunctivaewas transmitted between domestic and wild animals that were grazing in proximate pastures. Certain animals also presented infections with two different strains simultaneously.
Effect of Temperature, pH, and Initial Cell Number on
luxCDABE
and
nah
Gene Expression during Naphthalene and Salicylate Catabolism in the Bioreporter Organism
Pseudomonas putida
RB1353
Authors: Dorn Jonathan G. , Frye Robert J. , Maier Raina M. 
Published: April 2003

ABSTRACT

One limitation of employing
lux
bioreporters to monitor in situ microbial gene expression in dynamic, laboratory-scale systems is the confounding variability in the luminescent responses. For example, despite careful control of oxygen tension, growth stage, and cell number, luminescence from
Pseudomonas putida
RB1353, a naphthalene-degrading
lux
bioreporter, varied by more than sevenfold during saturated flow column experiments in our laboratory. Therefore, this study was conducted to determine what additional factors influence the luminescent response. Specifically, this study investigated the impact of temperature, pH, and initial cell number (variations within an order of magnitude) on the peak luminescence of
P. putida
RB1353 and the maximum degradation rate (
V
max
) during salicylate and naphthalene catabolism. Statistical analyses based on general linear models indicated that under constant oxygen tension, temperature and pH accounted for 98.1% of the variability in luminescence during salicylate catabolism and 94.2 and 49.5% of the variability in
V
max
during salicylate and naphthalene catabolism, respectively. Temperature, pH, and initial substrate concentration accounted for 99.9% of the variability in luminescence during naphthalene catabolism. Initial cell number, within an order of magnitude, did not have a significant influence on either peak luminescence or
V
max
during salicylate and naphthalene catabolism. Over the ranges of temperature and pH evaluated, peak luminescence varied by more than 4 orders of magnitude. The minimum parameter deviation required to alter
lux
gene expression during salicylate and naphthalene catabolism was a change in temperature of 1°C, a change in pH of 0.2, or a change in initial cell number of 1 order of magnitude. Results from this study indicate that there is a need for careful characterization of the impact of environmental conditions on both the expression of the reporter and catabolic genes and the activities of the gene products. For example, even though
lux
gene expression was occurring at ∼35°C, the luciferase enzyme was inactive. Furthermore, this study demonstrates that with careful characterization and standardization of measurement conditions, the attainment of a reproducible luminescent response and an understanding of the response are feasible.
Surface Display of the Receptor-Binding Region of the
Lactobacillus
brevis
S-Layer Protein in
Lactococcus
lactis
Provides Nonadhesive Lactococci with the Ability To Adhere to Intestinal Epithelial Cells
Authors: Åvall-Jääskeläinen Silja , Lindholm Agneta , Palva Airi 
Published: April 2003

ABSTRACT

Lactobacillus
brevis
is a promising lactic acid bacterium for use as a probiotic dietary adjunct and a vaccine vector. The N-terminal region of the S-layer protein (SlpA) of
L
.
brevis
ATCC 8287 was recently shown to mediate adhesion to various human cell lines in vitro. In this study, a surface display cassette was constructed on the basis of this SlpA receptor-binding domain, a proteinase spacer, and an autolysin anchor. The cassette was expressed under control of the
nisA
promoter in
Lactococcus
lactis
NZ9000. Western blot assay of lactococcal cell wall extracts with anti-SlpA antibodies confirmed that the SlpA adhesion domain of the fusion protein was expressed and located within the cell wall layer. Whole-cell enzyme-linked immunosorbent assay and immunofluorescence microscopy verified that the SlpA adhesion-mediating region was accessible on the lactococcal cell surface. In vitro adhesion assays with the human intestinal epithelial cell line Intestine 407 indicated that the recombinant lactococcal cells had gained an ability to adhere to Intestine 407 cells significantly greater than that of wild-type
L
.
lactis
NZ9000. Serum inhibition assay further confirmed that adhesion of recombinant lactococci to Intestine 407 cells was indeed mediated by the N terminus-encoding part of the
slpA
gene. The ability of the receptor-binding region of SlpA to adhere to fibronectin was also confirmed with this lactococcal surface display system. These results show that, with the aid of the receptor-binding region of the
L
.
brevis
SlpA protein, the ability to adhere to gut epithelial cells can indeed be transferred to another, nonadhesive, lactic acid bacterium.
Functional Replacement of the
Escherichia coli
d-
(−)-Lactate Dehydrogenase Gene (
ldhA
) with the
l-
(+)-Lactate Dehydrogenase Gene (
ldhL)
from
Pediococcus acidilactici
Authors: Zhou Shengde , Shanmugam K. T. , Ingram L. O. 
Published: April 2003

ABSTRACT

The microbial production of
l-
(+)-lactic acid is rapidly expanding to allow increased production of polylactic acid (PLA), a renewable, biodegradable plastic. The physical properties of PLA can be tailored for specific applications by controlling the ratio of
l-
(+) and
d
-(−) isomers. For most uses of PLA, the
l-
(+) isomer is more abundant. As an approach to reduce costs associated with biocatalysis (complex nutrients, antibiotics, aeration, product purification, and waste disposal), a recombinant derivative of
Escherichia coli
W3110 was developed that contains five chromosomal deletions (
focA-pflB frdBC adhE ackA ldhA
). This strain was constructed from a
d
-(−)-lactic acid-producing strain, SZ63 (
focA-pflB frdBC adhE ackA
), by replacing part of the chromosomal
ldhA
coding region with
Pediococcus acidilactici ldhL
encoding an
l
-lactate dehydrogenase. Although the initial strain (SZ79) grew and fermented poorly, a mutant (SZ85) was readily isolated by selecting for improved growth. SZ85 exhibited a 30-fold increase in
l
-lactate dehydrogenase activity in comparison to SZ79, functionally replacing the native
d
-lactate dehydrogenase activity. Sequencing revealed mutations in the upstream, coding, and terminator regions of
ldhL
in SZ85, which are presumed to be responsible for increased
l
-lactate dehydrogenase activity. SZ85 produced
l
-lactic acid in M9 mineral salts medium containing glucose or xylose with a yield of 93 to 95%, a purity of 98% (based on total fermentation products), and an optical purity greater than 99%. Unlike other recombinant biocatalysts for
l
-lactic acid, SZ85 remained prototrophic and is devoid of plasmids and antibiotic resistance genes.
Compatibility of Rhizobial Genotypes within Natural Populations of
Rhizobium leguminosarum
Biovar viciae for Nodulation of Host Legumes
Authors: Laguerre Gisèle , Louvrier Philippe , Allard Marie-Reine , Amarger Noëlle 
Published: April 2003

ABSTRACT

Populations of
Rhizobium leguminosarum
biovar viciae were sampled from two bulk soils, rhizosphere, and nodules of host legumes, fava bean (
Vicia faba
) and pea (
Pisum sativum
) grown in the same soils. Additional populations nodulating peas, fava beans, and vetches (
Vicia sativa
) grown in other soils and fava bean-nodulating strains from various geographic sites were also analyzed. The rhizobia were characterized by repetitive extragenomic palindromic-PCR fingerprinting and/or PCR-restriction fragment length polymorphism (RFLP) of 16S-23S ribosomal DNA intergenic spacers as markers of the genomic background and PCR-RFLP of a nodulation gene region,
nodD
, as a marker of the symbiotic component of the genome. Pairwise comparisons showed differences among the genetic structures of the bulk soil, rhizosphere, and nodule populations and in the degree of host specificity within the Vicieae cross-inoculation group. With fava bean, the symbiotic genotype appeared to be the preponderant determinant of the success in nodule occupancy of rhizobial genotypes independently of the associated genomic background, the plant genotype, and the soil sampled. The interaction between one particular rhizobial symbiotic genotype and fava bean seems to be highly specific for nodulation and linked to the efficiency of nitrogen fixation. By contrast with bulk soil and fava bean-nodulating populations, the analysis of pea-nodulating populations showed preferential associations between genomic backgrounds and symbiotic genotypes. Both components of the rhizobial genome may influence competitiveness for nodulation of pea, and rhizosphere colonization may be a decisive step in competition for nodule occupancy.
Heavy Metal Resistance of Biofilm and Planktonic
Pseudomonas aeruginosa
Authors: Teitzel Gail M. , Parsek Matthew R. 
Published: April 2003

ABSTRACT

A study was undertaken to examine the effects of the heavy metals copper, lead, and zinc on biofilm and planktonic
Pseudomonas aeruginosa
. A rotating-disk biofilm reactor was used to generate biofilm and free-swimming cultures to test their relative levels of resistance to heavy metals. It was determined that biofilms were anywhere from 2 to 600 times more resistant to heavy metal stress than free-swimming cells. When planktonic cells at different stages of growth were examined, it was found that logarithmically growing cells were more resistant to copper and lead stress than stationary-phase cells. However, biofilms were observed to be more resistant to heavy metals than either stationary-phase or logarithmically growing planktonic cells. Microscopy was used to evaluate the effect of copper stress on a mature
P. aeruginosa
biofilm. The exterior of the biofilm was preferentially killed after exposure to elevated concentrations of copper, and the majority of living cells were near the substratum. A potential explanation for this is that the extracellular polymeric substances that encase a biofilm may be responsible for protecting cells from heavy metal stress by binding the heavy metals and retarding their diffusion within the biofilm.
The Surface-Associated and Secreted MopE Protein of
Methylococcus capsulatus
(Bath) Responds to Changes in the Concentration of Copper in the Growth Medium
Authors: Karlsen Odd A. , Berven Frode S. , Stafford Graham P. , Larsen Øivind , Murrell J. Colin , Jensen Harald B. , Fjellbirkeland Anne 
Published: April 2003

ABSTRACT

Expression of surface-associated and secreted protein MopE of the methanotrophic bacterium
Methylococcus capsulatus
(Bath) in response to the concentration of copper ions in the growth medium was investigated. The level of protein associated with the cells and secreted to the medium changed when the copper concentration in the medium varied and was highest in cells exposed to copper stress.
Wide Geographic Distribution of Bacteriophages That Lyse the Same Indigenous Freshwater Isolate (
Sphingomonas
sp. Strain B18)
Authors: Wolf Arite , Wiese Jutta , Jost Günter , Witzel Karl-Paul 
Published: April 2003

ABSTRACT

An indigenous freshwater bacterium (
Sphingomonas
sp. strain B18) from Lake Pluβsee (Schleswig-Holstein, Germany) was used to isolate 44 phages from 13 very different freshwater and brackish habitats in distant geographic areas. This bacterial strain was very sensitive to a broad spectrum of phages from different aquatic environments. Phages isolated from geographically distant aquatic habitats, but also those from the same sample, were diverse with respect to morphology and restriction pattern. Some phages were widely distributed, while different types coexisted in the same sample. It was concluded that phages could be a major factor in shaping the structure of bacterial communities and maintaining a high bacterial diversity.
Manipulation of the phenotype of immortalised rat hepatocytes by different culture configurations and by dimethyl sulphoxide
Authors: Grant M I , Anderson K , McKay G , Wills M , Henderson C , MacDonald C 
Published: April 2003

The liver-specific phenotype of immortalised rat hepato-cytes is not irretrievably lost as they age in culture but can be manipulated by modifying the culture environment. Testosterone metabolism was used to investigate the profile of cytochrome P450 isoenzymes present in two immortalised cell lines, P9 and LQC, and in primary cultures of rat hepatocytes, cultured on collagen films, gels and double gel cultures (sandwich configuration). The extent of testosterone metabolism, and the range of metabolites produced, was increased in immortalised cells by the presence of collagen as a substratum film or gel but survival was poorer and the range of metabolites was reduced in sandwich culture. In contrast, testosterone metabolism was retained in primary hepatocytes in sandwich cultures at a higher level than in collagen film or gel cultures. Expression of alpha class glutathione-S-transferases (GSTs) increased and that of GSTP1 decreased (changes which indicate a recovery of normal liver GST phenotype) when the medium of immortalised cell cultures was supplemented with dimethyl sulph-oxide (DMSO). DMSO also improved ethoxyresorufin 0-deethylation (EROD) and testosterone metabolism in immortalised cells. It also markedly inhibited prolifera-tion, DNA, RNA and protein synthesis. Maximal testosterone metabolism was observed in immortalised cells cultured on collagen gels in the presence of l1o (v/v) DMSO. Development of a protocol for treating immortalised liver cells cultured on collagen gels with DMSO to switch between proliferation and differentiation may provide a convenient system expres-sing the xenobiotic metabolising enzymes required for in vitro toxicity testing.
Rat alveolar and interstitial macrophages in the fibrosing stage following quartz exposure
Authors: Zetterberg G , Elmberger G , Johansson A , Lundahl J , Lundborg M , Sköld C M , Tornling G , Camner P , Eklund A 
Published: April 2003

Exposure to quartz induces pulmonary inflammation and development of fibrosis. In order to study the fibrosing process, we investigated morphology function and phenotype of alveolar (AMs) and interstitial (IMs) macrophages at an early stage of fibrosis in rats. Rats were exposed by intratracheal instillations of 10 mg quartz (n = 8) or saline (n = 8) and studied 3 months later. AMs were obtained by bronchoalveolar lavage and IMs by mechanical fragmentation, followed by enzymatic digestion of lung tissue. Histology revealed subacute silicosis, with early focal fibrosis and alveolar lipoproteinosis. AM quartz exposure increased phagocytic activity and expression of major histocompatibility complex (MHC) Ia antigens, the latter being associated with cellular antigen presenting capacity. IM had an even more pronounced expression of MHC than AM after quartz exposure. Both macrophage fractions had a higher expression of OX-42 (complement receptor 3, CR3) than controls, but the increase in the IM fraction might be explained by the remaining AM in the IM fraction. Exposed AM adhered less to extracellular matrix components (vitronectin and fibronectin) than controls. In contrast, the adhesion of IM to vitronectin increased after exposure. Besides increased adhesion, the effects on IM were scarce. Our results therefore do not support the hypothesis that IM has a key role in the process of inflammation, including fibrosis.
In vitro and in vivo studies on chelation of manganese
Authors: Missy P , Lanhers M-C , Grignon Y , Joyeux M , Burnel D 
Published: April 2003

This work deals with new chelating agents of manganese (Mn). Out of 24 compounds chosen for their chemical structure supposed to be favorable for Mn complexation, six polyaminopolycarboxylic acids proved to be efficient for displacing Mn bound to serum bovine proteins in vitro: TTHA, DTPA, DPTA, DPTA – OH, HBED, EDTA (mobilization>50%). The first five compounds were then tested in vivo on rats pretreated with MnC12. They exhibited only slight to moderate efficacy to diminish Mn in tissues and were ineffective on increased Mn concentration in whole blood; in addition, they had different and specific mobilizing effects on other essential elements (Fe, Zn, Cu). Their limited efficacyin vivo could be due to the formation of very stable complexes between Mn2+ and different molecules such as hemoglobin and certain cytochromes, instead of Fe2+. This could disturb the functioning of the cellular respiratory chain, leading to an incomplete reduction of 02 with formation of free oxygenated radicals, reduction in the energy supply, and disturbance of the cytochromes renewal mechanism. All of these phenomena could accelerate cellular aging and explain the lack of efficacy ofthe chelating agents towards Mn neurotoxicity (Parkinson's syndrome).
Aggressive atropinisation and continuous pralidoxime (2-PAM) infusion in patients with severe organophosphate poisoning: experience of a northwest Indian hospital
Authors: Singh S , Chaudhry D , Behera D , Gupta D , Jindal S K 
Published: April 2003

OBJECTIVE: The aim of the study was to find whether continuous pralidoxime (2-PAM) infusion along with aggressive atropinisation improves the outcome in patients with severe organophosphate poisoning who require assisted ventilation. METHODS: Sixteen patients admitted to the respiratory intensive care unit (RICU) with severe organophosphate poisoning and requiring assisted ventilation were included in the study. The compounds involved were phorate (six), dichlorvos (four), oxydimeton methyl (one), monocrotophos (one), methyl parathion (one) and in three it was unknown. After decontamination, they were given intravenous (iv) bolus atropine 5 mg at onset and then 2.5 mg every 5-10 min till atropinisation was achieved, and then maintained either by intermittent bolus doses or by continuous infusion if the required dose was large. They were also given continuous iv infusion of 2-PAM in dose of 7.5 mg/kg body weight/h (maximum 500 mg/h) after an initial bolus dose of 2 g. RESULTS: The mean (± S.D.) dose of atropine was 735.02 ± 742.98 mg (range 85-3000 mg) with maximum dose on day 1. The mean (± S.D) duration of2-PAM infusion was 96.4±49.4 h (range 10-216 h). The mean (±S.D) duration of mechanical ventilation (MV) was 131.5 ± 95.65 h (range 4-336 h). Fourteen patients could be successfully extubated and two died of bronchopneumonia and sepsis (mortality 12.5%6). CONCLUSION: Continuous 2-PAM infusion along with aggressive atropinisation after initial decontamination improved the outcome but notthe duration of MV in severely intoxicated patients with organophosphate compounds who required assisted ventilation in this case series.
Immunotoxicological investigations on rats treated subacutely with dimethoate, As3+and Hg2+ in combination
Authors: Institóris L , Siroki O , Ündeger Ü , Basaran N , Dési I 
Published: April 2003

Effects of combined exposure with dimethoate (DM), HgCl2(Hg),andNaAsO2 (As)wereinvestigatedfollowinga28-day oral exposure in male Wistar rats. In preliminary experiments, the LOEL (Lowest Observed Effect Level) and NOEL (Non Observed Effect Level) doses of the substances were determined using the same experimental system [determination of body weight gain, organ weights of brain, thymus, heart, lung, kidneys, adrenals, spleen, testicles, popliteal lymph node, white blood cell (WBC) and red blood cell (RBC) count, haematocrit (Ht), mean cell volume (MCV) of RBCs, cell content of the femoral bone marrow, IgM-plaque forming cell (PFC) content of the spleen, delayed type hypersensitivity (DTH) reaction] and animal strain. In the combination studies, LOEL dose of DM (28.2 mg/kg) was combinedwithNOELdosesoftheheavymetals(HgCl2 =0.40 mg/kg, NaAsO2 =3.33 mg/kg), and vice versa (DM=7.04 mg/kg, HgCl2 =3.20 mg/kg, NaAsO2 =13.3 mg/kg). In the DM–Hg combinations, significant alterations were found versus the corresponding high-dose internal control in the body weight gain, relative liver and kidney weights, and in the PFC response. When DM was combined with As, interactions were indicated by changes of relative liver weight, MCV value, and the PFC content of the spleen. These results support the theory that the interactions between pesticides and heavy metals may modify the toxic effects of the single substances, and may also change the functional detection limits of the exposure. The changes in the functional detection limits, if they occur, can lead to false-positive and false-negative results in human epidemiological studies.