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A framework for risk characterization of environmental pollutants.

Risk characterization is defined by both the U.S. National Academy of Sciences and the U.S. EPA as the estimation of human health risk due to harmful (i.e., toxic or carcinogenic) substances or organisms. Risk characterization studies are accomplished by integrating quantitative exposure estimates and dose-response relationships with the qualitative results of hazard identification. A Risk Characterization Framework has been developed to encourage a systematic approach for analysis and presentation of risk estimates. This methodology subdivides the four common components of the risk assessment process into ten elements. Each of these elements is based on a term in a predictive risk equation. The equation allows independent computations of exposure, dose, lifetime individual risk, and risk to affected populations. All key assumptions in the predictive risk equation can be explicitly shown. This is important to understand the basis and inherent uncertainties of the risk estimation process. The systematic treatment of each of the ten elements in this framework aids in the difficult job of comparing risk estimates by different researchers using different methodologies. The Risk Characterization Framework has been applied to various indoor and outdoor air pollutants of a carcinogenic nature. With further development, it also promises to be applicable to noncarcinogenic effects.

Air Pollution, Indoor↗

Characterization of indoor particle sources using continuous mass and size monitors.

A comprehensive indoor particle characterization study was conducted in nine Boston-area homes in 1998 in order to characterize sources of PM in indoor environments. State-of-the-art sampling methodologies were used to obtain continuous PM2.5 concentration and size distribution particulate data for both indoor and outdoor air. Study homes, five of which were sampled during two seasons, were monitored over week-long periods. Among other data collected during the extensive monitoring efforts were 24-hr elemental/organic carbon (EC/OC) particulate data as well as semi-continuous air exchange rates and time-activity information. This rich data set shows that indoor particle events tend to be brief, intermittent, and highly variable, thus requiring the use of continuous instrumentation for their characterization. In addition to dramatically increasing indoor PM2.5 concentrations, these data demonstrate that indoor particle events can significantly alter the size distribution and composition of indoor particles. Source event data demonstrate that the impacts of indoor activities are especially pronounced in the ultrafine (da < or = 0.1 micron) and coarse (2.5 < or = da < or = 10 microns) modes. Among the sources of ultrafine particles characterized in this study are indoor ozone/terpene reactions. Furthermore, EC/OC data suggest that organic carbon is a major constituent of particles emitted during indoor source events. Whether exposures to indoor-generated particles, particularly from large short-term peak events, may be associated with adverse health effects will become clearer when biological mechanisms are better known.

Air Pollution, Indoor↗

Novel matrix invariants for characterization of changes of proteomics maps.

Previous studies on mathematical characterization of proteomics maps by sets of map invariants were based on the construction of a set of distance-related matrices obtained by matrix multiplication of a single matrix by itself. Here we consider an alternative characterization of proteomics maps based on a set of matrices characterizing local features of an embedded zigzag curve over the map. It is shown that novel invariants can well characterize proteomics maps. Advantages of the novel approach are discussed.

Models, Theoretical↗

The early stages of amyloid formation: biophysical and structural characterization of human calcitonin pre-fibrillar assemblies.

Amyloid fibril formation is a nucleation dependent process characterized by a lag-phase prior to the appearance of detectable amyloid fibrils. While the three-dimensional structure of amyloid fibrils at atomic resolution is just beginning to be elucidated, the early process of monomers assembly into oligomers is less understood. Understanding the dynamic processes that lead to the formation of these intermediates is highly important as these assemblies might be the most pathological ones. Here, we investigated the biophysical and structural features characterizing the early stage assemblies formed by the human hormone calcitonin. We calculated the initial nucleus size by experimentally determining the dependence between the lag-time length and the hCT concentrations. We used size exclusion chromatography and dynamic light scattering in order to characterize the dynamic growth process of preliminary intermediates transformed into larger structures. The early structures were visualized using high-resolution transmission electron microscopy. Annular pore-like structures were observed along with protofibrilar structures. This observed morphology is similar to structures revealed during the fibrillization processes of beta-amyloid, alpha-synuclein, and islet amyloid polypeptide, suggesting that these intermediates represent a generic early structure conformation. The results introduced here imply that a variety of intermediate assemblies are formed during the early stages of amyloid fibril formation. The characterizing of their structural features and assembly kinetics will contribute to the rational design of inhibitors directed towards early structure assemblies.

Amyloid↗

A web management service applied to a comprehensive characterization of Visible Human Dataset colour images.

Visible Human Dataset (VHD) is a remarkable piece of raw digital anatomical knowledge still to be fully exploited. Colours of VHD anatomic images are the natural targets of different algorithmic approaches devoted to understanding the content of the complex digital medical images, but they have never been analysed exhaustively. A full colorimetric characterization of all 9000 VHD colour images may help to take advantage of implicit available information in raw data. This study describes a novel colorimetric characterization and a Visual Knowledge Discovery tool, using methods from database field, data visualization, and image analysis. The applied heterogeneous methods allowed us to develop a histogram meta database and make it available remotely. It consists of a histogram-based colorimetric characterization of the all VHD 24-bit colour images. A user-friendly, interactive, and intuitive 3D framework providing 3D services was built and made freely available. It allows real-time analysis of colour component characteristics of a user-defined set of VHD images providing 3D interactive navigation of the histogram meta database. New knowledge can be discovered using our tool and the histogram meta database provided. This work allowed us to propose novel methods for colour image characterization and obtained results using developed service on VHD colour images let us to partially understand the not fully satisfactorily results achieved so far analysing these images.

Anatomy↗

Physical and chemical characterization of beryllium particles from several workplaces in Québec, Canada--part A: determining methods for the analysis of low levels of beryllium.

Chemical and physical characterizations of beryllium (Be) particles found in settled dust samples from four industries based in Québec were attempted using a variety of analytical methods. Bulk particle chemistry was determined using inductively coupled plasma-mass spectrometry (ICP-MS), graphite furnace atomic absorption spectrometry (GFAAS), and instrumental neutron activation analysis (INAA). Time-of-flight secondary-ion mass spectrometry (TOF-SIMS), transmission electron microscopy, scanning electron microscopy, energy-dispersive spectroscopy, x-ray diffraction (XRD), electron energy loss spectrometry (EELS), and Auger microscopy were used to characterize physicochemical properties of particles. These analyses were deemed important based on the hypotheses that (1) different chemical forms of Be do not present the same risks, and (2) different morphologies lead to different risks. Standards were used to prove the adequacy of XRD, EELS, and Auger microscopy prior to the analyses of industrial samples. However, low concentrations of Be in samples were a limiting factor for most methods; few detected Be in industrial samples. Only ICP-MS, GFAAS, and TOF-SIMS were able to detect Be in industrial samples analyzed in this study. Characterization of settled dust samples showed high number of Be particles, even for Be concentrations below 100 ppm. Furthermore, Be seems to be present as fine particles of Be metal, possibly mechanically agglomerated or aggregated to larger particles or compounds such as cryolite. Other major elements detected with INAA present in the samples were limited to Na, Al, Ca, and F. It was concluded that TOF-SIMS is a valid method for characterizing particles containing approximately 0.01% Be.

Beryllium↗

Production and characterization of an anti-idiotypic single chain Fv that recognizes an anti-DNA antibody.

A well-characterized recombinant anti-idiotype to an anti-DNA antibody can be useful for studies of the regulation of anti-DNA-producing B cells. Using a hybridoma technique, a monoclonal anti-idiotypic antibody, designated O2F3, was obtained, and its scFv gene was constructed. O2F3 single chain Fv (scFv) was produced against an idiotope of a monoclonal anti-DNA antibody, 3D8, that was obtained from an autoimmune-prone mouse, MRL-lpr/lpr. Here we describe the production and in vitro characterization of the O2F3 scFv, and compare it with its parent monoclonal antibody, O2F3 IgM. To characterize O2F3 scFv and O2F3 IgM, we generated recombinant 3D8 fragments, including 3D8 scFv, 3D8 VH, and 3D8 VL, that were used as antigens in several assays. ELISA and Western blot analysis showed that both O2F3 scFv and O2F3 IgM recognized a conformational determinant formed by the association of the variable region heavy and light chains of the 3D8 antibody, suggesting that O2F3 scFv retained a similar binding pattern to its parent O2F3 antibody. The idiotope recognized by O2F3 was shown by competitive ELISA to be outside of the DNA binding site of the 3D8 antibody. This characterized O2F3 scFv could be applied for the regulation of anti-DNA antibody production and the manipulation of recombinant antibody-based proteins to which toxins, enzymes, and chemical agents can be connected.

Animals↗

Molecular characterization of methicillin-resistant coagulase-negative staphylococci from a neonatal intensive care unit.

OBJECTIVE: To evaluate clonal dissemination of methicillin-resistant coagulase-negative staphylococci (CNS). SETTING: Neonatal intensive care unit of a 180-bed, university-affiliated general hospital. PATIENTS: Neonates admitted to the neonatal intensive care unit between March 1999 and October 2000, from whom CNS were isolated as a unique pathogen. Patients from other wards from whom epidemiologically unrelated staphylococci strains were obtained served as control-patients. METHODS: Conventional methods were used for phenotypic characterization of CNS. Methicillin resistance was determined by mecA polymerase chain reaction (PCR) amplification. Genotypic characterization was done by random amplification of DNA with degenerated primers (RAPD) and repetitive element sequence-based PCR (rep-PCR). RESULTS: Forty methicillin-resistant CNS isolates obtained from neonates were characterized as Staphylococcus epidermidis (33), S. hominis (5), S. warneri (1), and S. auricularis (1). Both RAPD and rep-PCR indicated the presence of 4 different clones among the 33 S. epidermidis isolates. In turn, the 4 randomly selected, epidemiologically unrelated methicillin-resistant CNS strains obtained from control-patients showed 3 new profiles by RAPD and 2 by rep-PCR, which differed from the corresponding patterns mentioned earlier. Persistence of S. hominis in a neonate could be assessed by both genotypic techniques. CONCLUSIONS: The molecular characterization of the methicillin-resistant CNS studied indicated dissemination of one particular methicillin-resistant CNS clone among the neonates in the ward studied. Although RAPD showed a superior power to discriminate among methicillin-resistant CNS isolates, both RAPD and rep-PCR detected intraspecific and interspecific genomic diversity.

Argentina↗

In vivo localized 1H NMR spectroscopy: an experimental characterization of the PRESS technique.

The aim of this study was the experimental characterization of the PRESS technique for image-guided in vivo 1H NMR spectroscopy, as implemented on a commercially available 1.5 T NMR whole-body system. Three characterization parameters were defined and evaluated in test object measurements: selection efficiency, suppression efficiency and contamination. The characterization parameters were evaluated in different experimental conditions. The profile of signal intensity across the volume of interest (VOI) was investigated. Our results show that the characterization parameters are slightly affected by the echo time, while they are not affected by changes in the repetition time. When the VOI is co-extensive with the inner cube of the test object the suppression efficiency is good but the selection efficiency is not very high. Contamination is affected by the synthesizer frequency offset, while the selection efficiency is quite stable. Contamination effects become significant in case of 90 degrees pulse maladjustment.

Contrast Media↗

Near full-length genome characterization of an HIV type 1 CRF05_DF virus from Spain.

We report the near full-length sequence characterization of a HIV-1 DF intersubtype recombinant virus from Spain, X492, directly amplified from peripheral blood mononuclear cells' DNA. This isolate shares an identical mosaic structure and exhibits consistent phylogenetic clustering along the genome with VI961, a previously characterized DF recombinant virus. By contrast, VI1310, which may represent the same recombinant form as VI961 (CRF05_DF), is only partially homologous to VI961 and X492. Of three additional DF recombinant viruses previously characterized in gag-pol, only one, VI1267, clusters uniformly with VI961 and X492; the other two branch separately in a segment of pol. These results allow us to define an HIV-1 circulating recombinant form (CRF05_DF), characterized in near full-length genomes of two isolates (VI961 and X492) and in partial gag-pol sequences of a third virus (VI1267). Three other reported DF recombinant viruses, including the fully sequenced VI1310, exhibit incomplete homology to VI961 and X492.

Genome, Viral↗

Genomes to Life "Center for Molecular and Cellular Systems": a research program for identification and characterization of protein complexes.

Goal 1 of Department of Energy's Genomes to Life (GTL) program seeks to identify and characterize the complete set of protein complexes within a cell. Goal 1 forms the foundation necessary to accomplish the other objectives of the GTL program, which focus on gene regulatory networks and molecular level characterization of interactions in microbial communities. Together this information would allow cells and their components to be understood in sufficient detail to predict, test and understand the responses of a biological system to its environment. The Center for Molecular and Cellular Systems has been established to identify and characterize protein complexes using high through-put analytical technologies.A dynamic research program is being developed that supports the goals of the Center by focusing on the development new capabilities for sample preparation and complex separations, molecular level identification of the protein complexes by mass spectrometry, characterization of the complexes in living cells by imaging techniques, and bioinformatics and computational tools for the collection and interpretation of data and formation of databases and tools to allow the data to be shared by the biological community.

Animals↗

Why the NOSPECS classification of Graves' eye disease should be abandoned, with suggestions for the characterization of this disease.

The NOSPECS classification of Graves' eye disease is inadequate for a number of reasons: (a) poor characterization of the condition with no indication of disease activity, marked underrepresentation of eyelid position, and overrepresentation of corneal problems, (b) it represents neither a continuous nor stepladder progression of disease as the numbered classification would suggest, (c) parts of it are subjective, and (d) the gradings of classes 3, 5, and 6 are poor. Graves' eye disease should be characterized rather than classified, using objective clinical assessments of optic nerve function, proptosis, ocular motility, and eyelid position. It is recommended that a multicenter clinical trial be done to determine the significance of different values and changes in values for each assessment. Disease activity is to be defined by the clinical trial in terms of changes of the four assessments as a function of time. This should produce a simple characterization that would create an image of the patient's eye problems for a knowledgeable physician and be sufficient to guide decisions on whether currently accepted surgical therapies are advisable. Data from the clinical trial would enable the characterization to predict the likelihood of success of various immunosuppressive therapies.

Graves Disease↗

Morphological and secretory characterization of extrafloral nectaries in plants of coastal Veracruz, Mexico.

BACKGROUND AND AIMS: Morphological descriptions of the extrafloral nectaries (EFNs) of certain plant species are common in the literature, but they rarely relate morphology with histology, gland distribution and secretory attributes. In this study a morphological/secretory characterization of EFNs occurring on several plant species in a tropical coastal community is made and the implications of gland attributes discussed from a functional perspective. METHODS: The morphology and nectar secretion of the EFNs of 20 plant species are characterized through scanning electron microscopy, histochemical detection of reducing sugars (Fehling's reagent) and nectar volume/concentration estimates. KEY RESULTS: Sixty-five per cent of plant species in coastal communities had EFNs on vegetative structures and 35 % of species had glands on reproductive and vegetative organs. The Fabaceae is the plant family with the most species with EFNs and most diversity of gland morphologies. Four types of vascularized nectaries and four of glandular trichomes are described; sugar-secreting trichomes are characterized using Fehling's technique, and the first descriptions of unicellular and peltate trichomes functioning as EFNs are provided. Glands of ten plant species and six genera are described for the first time. Four plant species possess more than one morphological type of EFN. Eleven species have EFNs in more than one location or organ. More complex glands secrete more nectar, but are functionally homologous to the aggregations of numerous secretory trichomes on specific and valuable plant organs. CONCLUSION: Important diversity of EFN morphology was foundin the coastal plant community studied. Both vascularized and non-vascularized EFNs are observed in plants and, for the latter, previously non-existent morpho-secretory characterizations are provided with a methodological approach to study them. It is recommended that studies relating EFN attributes (i.e. morphology, distribution) with their differential visitation by insects (i.e. ants) and the cost of maintenance to the plants are carried out to understand the evolution of these glands.

Acacia↗

32P-HPLC suitable for characterization of DNA adducts formed in vitro by polycyclic aromatic hydrocarbons and derivatives.

Analysis of DNA adducts demands both high sensitivity and good resolution. A high-performance liquid chromatography method for 32P-postlabeled DNA adducts (32P-HPLC) was used to investigate DNA adduct formation from 38 polycyclic hydrocarbons and biphenyls in vitro. The 32P-HPLC method proved to be useful for separation, detection and characterization of DNA adducts from most of the substances. The in vitro method used to form the DNA adducts, with calf thymus DNA, nucleotide 3'-phosphates and metabolic activation through S-9 liver homogenate, gave poor quantitative reproducibility. However, the results showed that the 32P-HPLC method was suitable for characterizing DNA adducts from many substances. From 35 of the tested substances 365 DNA and nucleotide 3'-phosphate adducts were detected and characterized concerning retention times. Of the adducts, 171 were detected in DNA and 39 of them from five substances were characterized concerning target nucleotides. The retention time library built can be used in future analyses of DNA with complex patterns of DNA adducts.

Animals↗

Cloning and characterization of the scalloped region of Drosophila melanogaster.

Viable mutants of the scalloped gene (sd) of Drosophila melanogaster exhibit defects that can include gapping of the wing margin and ectopic bristle formation on the wing. Lethal sd alleles characterized in the present study now implicate this gene in a genetic function essential for normal development. In order to further characterize the developmental role of this gene, we have undertaken to clone and characterize the region where sd maps. A P[ry+] transposon insertion at 13F associated with sd[ry+2216] served as the starting point for a 42-kb chromosomal walk. Molecular lesions associated with viable and lethal sd alleles were characterized by genomic hybridization analysis as a means of defining the extent of the gene. DNA rearrangements associated with 11 viable sd alleles map to a 2-kb interval which appears to be a "hot spot" for P element activity. Four of five recessive lethal sd mutations were mapped by denaturing gradient gel electrophoresis to a region 12-14 kb away from the region of viable lesions. In a sd+ genotype, at least two structurally related and developmentally regulated transcripts hybridize to the genomic region where several sd lethal alleles have been localized. A viable mutation, sd58, used for comparison in the transcript analysis, makes at least two slightly smaller transcripts that also hybridize to this region. Preliminary analysis of cDNA clones has identified three structurally related transcripts that hybridize to this genomic region. The 5' end of these transcripts extends into the 2-kb genomic region wherein DNA rearrangements were seen in the P element rearrangements. We favor the view that the transcripts represented by these cDNA clones are products of the sd gene. If this is true, the sd gene would include genomic sequences extending over at least 14 kb of the described chromosomal walk, and would appear to be subject to alternative splicing.

Alleles↗

Alpha 1,3 galactosyltransferase: new sequences and characterization of conserved cysteine residues.

Nucleotide sequences were determined for alpha1,3 galactosyltransferases (alpha1,3 GalTs) from several species (bat, mink, dog, sheep, and dolphin) and compared with those previously determined for this enzyme and members of the alpha1,3 galactosyl/N-acetylgalactosyltransferase (alpha1,3 Gal(NAc)Ts) family of enzymes. Sequence comparison of the newly characterized alpha1,3 GalT nucleotide and predicted amino acid sequences with those previously characterized for other alpha1,3GalT enzymes demonstrated a remarkable level of sequence identity at the nucleotide and amino acid level. The identity of each sequence as an alpha1,3 GalT was confirmed by expressing the encoded protein and characterizing the resulting enzyme. The alpha1,3 GalTs have a significant degree of sequence homology with A and B transferases, the alpha1,3 GalNAcT that catalyzes the synthesis of Forssman antigen, and the recently cloned iso-globotriaosylceramide synthase. Among the conserved residues, there are two Cys residues. To determine if these conserved residues are free or involved in the formation of a disulfide bond, bovine alpha1,3 GalT was characterized by chemical modification and mass spectrometry. Each peptide containing a Cys residue was chemically labeled with an alkylating reagent demonstrating that these enzymes do not contain disulfide bonds. Similar results have recently been reported for A and B transferases (Yen et al., 2000, J. Mass. Spectrom., 35, 990-1002). Thus, the highly conserved Cys residues found in these members of the alpha1,3 Gal(NAc)Ts family of enzymes are likely involved in other important aspects of enzyme structure/function within this enzyme family.

Amino Acid Sequence↗

Characterization of the human ABC superfamily: isolation and mapping of 21 new genes using the expressed sequence tags database.

As an approach to characterizing all human ATP-binding cassette (ABC) superfamily genes, a search of the human expressed sequence tag (EST) database was performed using sequences from known ABC genes. A total of 105 clones, containing sequences of potential ABC genes, were identified, representing 21 distinct genes. This brings the total number of characterized human ABC genes from 12 to 33. The new ABC genes were mapped by PCR on somatic cell and radiation hybrid panels and yeast artificial chromosomes (YACs). The genes are located on human chromosomes 1, 2, 3, 4, 6, 7, 10, 12, 13, 14, 16, 17 and X; at locations distinct from previously mapped members of the superfamily. The characterized genes display extensive diversity in sequence and expression pattern and this information was utilized to determine potential structural, functional and evolutionary relationships to previously characterized members of the ABC superfamily.

ATP-Binding Cassette Transporters↗

Characterization of a second human clathrin heavy chain polypeptide gene (CLH-22) from chromosome 22q11.

We report cloning and characterization of the second human clathrin heavy chain polypeptide gene (CLH-22) localized to chromosome 22q11. Hence H. sapiens is the first species for which two clathrin heavy chain genes have been reported. We provide 5470 bp cDNA sequence covering the entire open reading frame of the CLH-22 gene. The predicted polypeptide is composed of 1640 amino acids. Its 6 kb transcript is expressed in all of 16 tested human tissues, suggesting it is a housekeeping gene. Skeletal muscle, testis and heart show significantly higher expression levels. Compared to the previously characterized human clathrin heavy chain gene localized on chromosome 17 (CLH-17), CLH-22 shows different transcript size and expression profile in human tissues. Northern analysis of CLH-22 suggests that several alternatively spliced transcripts exist. A presumably single, 171 bp long alternatively spliced exon has been characterized. Amino acid sequence comparison between CLH-22 and CLH-17 shows an overall identify and similarity of 84.7 and 91.1%, respectively. At the nucleic acid level, identity between open reading frames of both genes is 74.3%. Sequence comparison with previously cloned genes in other species suggests that counterparts of the CLH-17 gene have been cloned in B. taurus and R. norvegicus, whereas presumptive mammalian homologues of the CLH-22 gene are yet to be characterized. Our Northern and Southern blot analyses of meningiomas clearly suggest the CLH-22 gene may be involved in the tumor development and can be considered as a candidate for a tumor suppressor.

Alternative Splicing↗