Biomedical subjects

Michael Berg

Publications and source records attributed to Michael Berg.

At least 19 recordsLinked to original sources

PubMed2006

Magnitude of arsenic pollution in the Mekong and Red River Deltas--Cambodia and Vietnam.

Large alluvial deltas of the Mekong River in southern Vietnam and Cambodia and the Red River in northern Vietnam have groundwaters that are exploited for drinking water by private tube-wells, which are of increasing demand since the mid-1990s. This paper presents an overview of groundwater arsenic pollution in the Mekong delta: arsenic concentrations ranged from 1-1610 microg/L in Cambodia (average 217 microg/L) and 1-845 microg/L in southern Vietnam (average 39 microg/L), respectively. It also evaluates the situation in Red River delta where groundwater arsenic concentrations vary from 1-3050 microg/L (average 159 microg/L). In addition to rural areas, the drinking water supply of the city of Hanoi has elevated arsenic concentrations. The sediments of 12-40 m deep cores from the Red River delta contain arsenic levels of 2-33 microg/g (average 7 microg/g, dry weight) and show a remarkable correlation with sediment-bound iron. In all three areas, the groundwater arsenic pollution seem to be of natural origin and caused by reductive dissolution of arsenic-bearing iron phases buried in aquifers. The population at risk of chronic arsenic poisoning is estimated to be 10 million in the Red River delta and 0.5-1 million in the Mekong delta. A subset of hair samples collected in Vietnam and Cambodia from residents drinking groundwater with arsenic levels >50 microg/L have a significantly higher arsenic content than control groups (<50 microg/L). Few cases of arsenic related health problems are recognized in the study areas compared to Bangladesh and West Bengal. This difference probably relates to arsenic contaminated tube-well water only being used substantially over the past 7 to 10 years in Vietnam and Cambodia. Because symptoms of chronic arsenic poisoning usually take more than 10 years to develop, the number of future arsenic related ailments in Cambodia and Vietnam is likely to increase. Early mitigation measures should be a high priority.

Arsenic↗

PubMed2006

Arsenite and arsenate binding to dissolved humic acids: influence of pH, type of humic acid, and aluminum.

The fate of arsenic in the aquatic environment is influenced by dissolved natural organic matter (DOM). Using an equilibrium dialysis method, conditional distribution coefficients (Dom) for As(III) and As(V) binding onto two commercial humic acids were determined at environmentally relevant As/dissolved organic carbon (DOC) ratios and as a function of pH. At all pH values, As(V) was more strongly bound than As(III). Maximum binding was observed around pH 7, which is consistent with H+ competition for binding sites at low pH values and OH- competition for the arsenic center at high pH. For both oxidation states, Dom values increased with decreasing As/DOC ratios. Dom values were fitted as a function of the As/DOC ratio for As(III) and As(V). Compared to the aquatic humic acid, the terrestrial humic acid had a higher affinity for arsenic binding with 1.5-3 times higher Dom values under the same conditions. Al3+ in excess to arsenic successfully competed for strong binding sites at low As/DOC ratios. Under environmentally relevant conditions, about 10% of total As(V) may be bound to DOM, whereas >10% of As(III) is bound to DOM at very low As/DOC ratios only. Binding of arsenic to DOM should be considered in natural systems.

Aluminum↗

PubMed2006

Arsenic removal from groundwater by household sand filters: comparative field study, model calculations, and health benefits.

Arsenic removal efficiencies of 43 household sand filters were studied in rural areas of the Red River Delta in Vietnam. Simultaneously, raw groundwater from the same households and additional 31 tubewells was sampled to investigate arsenic coprecipitation with hydrous ferric iron from solution, i.e., without contact to sand surfaces. From the groundwaters containing 10-382 microg/L As, < 0.1-48 mg/L Fe, < 0.01-3.7 mg/L P, and 0.05-3.3 mg/L Mn, similar average removal rates of 80% and 76% were found for the sand filter and coprecipitation experiments, respectively. The filtering process requires only a few minutes. Removal efficiencies of Fe, phosphate, and Mn were > 99%, 90%, and 71%, respectively. The concentration of dissolved iron in groundwater was the decisive factor for the removal of arsenic. Residual arsenic levels below 50 microg/L were achieved by 90% of the studied sand filters, and 40% were even below 10 microg/L. Fe/As ratios of > or = 50 or > or = 250 were required to ensure arsenic removal to levels below 50 or 10 microg/L, respectively. Phosphate concentrations > 2.5 mg P/L slightly hampered the sand filter and coprecipitation efficiencies. Interestingly, the overall arsenic elimination was higher than predicted from model calculations based on sorption constants determined from coprecipitation experiments with artificial groundwater. This observation is assumed to result from As(lll) oxidation involving Mn, microorganisms, and possibly dissolved organic matter present in the natural groundwaters. Clear evidence of lowered arsenic burden for people consuming sand-filtered water is demonstrated from hair analyses. The investigated sand filters proved to operate fast and robust for a broad range of groundwater composition and are thus also a viable option for mitigation in other arsenic affected regions. An estimation conducted for Bangladesh indicates that a median residual level of 25 microg/L arsenic could be reached in 84% of the polluted groundwater. The easily observable removal of iron from the pumped water makes the effect of a sand filter immediately recognizable even to people who are not aware of the arsenic problem.

Arsenic↗

PubMed2006

HPV16 L1 capsid protein expressed from viable adenovirus recombinants elicits neutralizing antibody in mice.

Immunization against human papillomavirus (HPV) infection promises to reduce the worldwide burden of cervical cancer. To evaluate the potential of live recombinant adenoviruses for induction of HPV infection-blocking immunity, we prepared viable adenovirus recombinants that express the HPV16 L1 gene from the adenovirus major late transcriptional unit. Adenovirus-produced HPV16 L1 assembles into virus-like particles (VLPs) in infected cells in culture. Purified HPV16 VLPs are recognized by HPV16 neutralizing antibodies and induce high neutralizing titers when injected intraperitoneally into mice. Canine oral papillomavirus VLPs derived from previously described recombinants also induce strong antibody responses in mice. These data support our suggestion that viable adenovirus recombinants will be able to induce protective immunity to papillomavirus infection during replication in human vaccinees.

Adenoviridae↗

PubMed2006

Kinetics and mechanistic aspects of As(III) oxidation by aqueous chlorine, chloramines, and ozone: relevance to drinking water treatment.

Kinetics and mechanisms of As(III) oxidation by free available chlorine (FAC-the sum of HOCl and OCl-), ozone (O3), and monochloramine (NH2Cl) were investigated in buffered reagent solutions. Each reaction was found to be first order in oxidant and in As(III), with 1:1 stoichiometry. FAC-As(III) and O3-As(III) reactions were extremely fast, with pH-dependent, apparent second-order rate constants, k''app, of 2.6 (+/- 0.1) x 10(5) M(-1) s(-1) and 1.5 (+/- 0.1) x 10(6) M(-1) s(-1) at pH 7, whereas the NH2Cl-As(III) reaction was relatively slow (k''app = 4.3 (+/- 1.7) x 10(-1) M(-1) s(-1) at pH 7). Experiments conducted in real water samples spiked with 50 microg/L As(III) (6.7 x 10(-7) M) showed that a 0.1 mg/L Cl2 (1.4 x 10-6 M) dose as FAC was sufficient to achieve depletion of As(III) to <1 microg/L As(III) within 10 s of oxidant addition to waters containing negligible NH3 concentrations and DOC concentrations <2 mg-C/L. Even in a water containing 1 mg-N/L (7.1 x 10(-5) M) as NH3, >75% As(III) oxidation could be achieved within 10 s of dosing 1-2 mg/L Cl2 (1.4-2.8 x 10(-5) M) as FAC. As(III) residuals remaining in NH3-containing waters 10 s after dosing FAC were slowly oxidized (t1/2 > or = 4 h) in the presence of NH2Cl formed by the FAC-NH3 reaction. Ozonation was sufficient to yield >99% depletion of 50 microg/L As(III) within 10 s of dosing 0.25 mg/L O3 (5.2 x 10(-6) M) to real waters containing <2 mg-C/L of DOC, while 0.8 mg/L O3 (1.7 x 10(-5) M) was sufficientfor a water containing 5.4 mg-C/L of DOC. NH3 had negligible effect on the efficiency of As(III) oxidation by O3, due to the slow kinetics of the O3-NH3 reaction at circumneutral pH. Time-resolved measurements of As(III) loss during chlorination and ozonation of real waters were accurately modeled using the rate constants determined in this investigation.

Ammonia↗

PubMed2005

Sequence comparisons of plasmids pBJS-O of Spiroplasma citri and pSKU146 of S. kunkelii: implications for plasmid evolution.

BACKGROUND: Spiroplasma citri BR3-3X and S. kunkelii CR2-3X cause serious diseases worldwide on citrus and maize species, respectively. S. citri BR3-3X harbors a plasmid, pBJS-Original (pBJS-O), that encodes the spiroplasma adhesion related protein 1 (SARP1), a protein implicated in binding of the pathogen to cells of its leafhopper vector, Circulifer tenellus. The S. kunkelii CR2-3X plasmid, pSKU146, encodes a homolog of SARP1, Sk-ARP1. Due to the close phylogenetic relationship of the two pathogens, we hypothesized that the two plasmids are closely related as well. RESULTS: The nucleotide sequence of pBJS-O was determined and compared to the sequences of a plasmid from BR3-T (pBJS-T), which is a multiply passaged leafhopper transmissible derivative of BR3-3X, and to known plasmid sequences including that of pSKU146. In addition to arp1, the 13,374 bp pBJS-O sequence putatively contains nine genes, recognized as open reading frames (ORFs). Several pBJS-O ORFs have homologs on pSKU146. However, the sequences flanking soj-like genes on both plasmids were found to be more distant from one another than sequences in any other region. Further, unlike pSKU146, pBJS-O lacks the conserved oriT region characteristic of the IncP group of bacterial plasmids. We were unable to identify a region in pBJS-O resembling a known plasmid origin of transfer. In regions where sequence was available for the plasmid from both BR3-3X and BR3-T, the pBJS-T sequence had a 0.4 kb deletion relative to its progenitor, pBJS-O. Southern blot hybridization of extrachromosomal DNA from various S. citri strains and spiroplasma species to an arp-specific probe and a probe made from the entire plasmid DNA of BR3-3X revealed limited conservation of both sequences in the genus Spiroplasma. Finally, we also report the presence on the BR3-3X chromosome of arp2, an S. citri homolog of arp1 that encodes the predicted protein SARP2. The C-terminal domain of SARP2 is homologous to that of SARP1, but its N-terminal domain is distinct. CONCLUSION: Our data suggest that pBJS is a novel S. citri plasmid that does not belong to any known plasmid incompatibility group. The differences between pBJS-O and pSKU146 suggest that one or more events of recombination have contributed to the divergence of the plasmids of the two sister Spiroplasma species; the plasmid from S. citri itself has diverged slightly during the derivation of S. citri BR3-T from BR3-3X. Our data also show that pBJS-O encodes the putative adhesin SARP1. The presence of traE and mob on pBJS-O suggests a role for the plasmid in spiroplasmal conjugation.

Adhesins, Bacterial↗

PubMed2005

Bacterial bioassay for rapid and accurate analysis of arsenic in highly variable groundwater samples.

In this study, we report the first ever large-scale environmental validation of a microbial reporter-based test to measure arsenic concentrations in natural water resources. A bioluminescence-producing arsenic-inducible bacterium based on Escherichia coli was used as the reporter organism. Specific protocols were developed with the goal to avoid the negative influence of iron in groundwater on arsenic availability to the bioreporter cells. A total of 194 groundwater samples were collected in the Red River and Mekong River Delta regions of Vietnam and were analyzed both by atomic absorption spectroscopy (AAS) and by the arsenic bioreporter protocol. The bacterial cells performed well at and above arsenic concentrations in groundwater of 7 microg/L, with an almost linearly proportional increase of the bioluminescence signal between 10 and 100 microg As/L (r2 = 0.997). Comparisons between AAS and arsenic bioreporter determinations gave an overall average of 8.0% false negative and 2.4% false positive identifications for the bioreporter prediction at the WHO recommended acceptable arsenic concentration of 10 microg/L, which is far betterthan the performance of chemical field test kits. Because of the ease of the measurement protocol and the low application cost, the microbiological arsenic test has a great potential in large screening campaigns in Asia and in other areas suffering from arsenic pollution in groundwater resources.

Arsenic↗

PubMed2005

Viable adenovirus vaccine prototypes: high-level production of a papillomavirus capsid antigen from the major late transcriptional unit.

Safe, effective, orally delivered, live adenovirus vaccines have been in use for three decades. Recombinant derivatives of the live adenovirus vaccines may prove an economical alternative to current vaccines for a variety of diseases. To explore that possibility, we constructed a series of recombinants that express the major capsid protein (L1) of canine oral papillomavirus (COPV), a model for mucosal human papillomavirus (HPV) infection. Vaccination with virus-like particles (VLPs) composed of recombinant HPV L1 completely prevents persistent HPV infection [Koutsky, L. A., Ault, K. A., Wheeler, C. M., Brown, D. R., Barr, E., Alvarez, F. B., Chiacchierini, L. M. & Jansen, K. U. (2002) N. Engl. J. Med. 347, 1645-1651], suggesting that L1 expressed from recombinant adenoviruses might provide protective immunity. In our recombinants, COPV L1 is incorporated into adenovirus late region 5 (Ad L5) and is expressed as a member of the adenoviral major late transcriptional unit (MLTU). COPV L1 production by the most prolific recombinant is comparable to that of the most abundant adenoviral protein, hexon. COPV L1 production by recombinants is influenced by Ad L5 gene order, the specific mRNA processing signals associated with COPV L1, and the state of a putative splicing inhibitor in the COPV L1 gene. Recombinant COPV L1 protein assembles into VLPs that react with an antibody specific for conformational epitopes on native COPV L1 protein that correlate with protection in vivo. The designs of these recombinants can be applied directly to the production of recombinants appropriate for assessing immunogenicity and protective efficacy in animal models and in human trials.

Adenoviridae↗

PubMed2005

New evaluation scheme for two-dimensional isotope analysis to decipher biodegradation processes: application to groundwater contamination by MTBE.

Compound-specific analysis of stable carbon and hydrogen isotopes was used to assess the fate of the gasoline additive methyl tert-butyl ether (MTBE) and its major degradation product tert-butyl alcohol (TBA) in a groundwater plume at an industrial disposal site. We present a novel approach to evaluate two-dimensional compound-specific isotope data with the potential to identify reaction mechanisms and to quantify the extent of biodegradation at complex field sites. Due to the widespread contaminant plume, multiple MTBE sources, the presence of numerous other organic pollutants, and the complex biogeochemical and hydrological regime atthe site, a traditional mass balance approach was not applicable. The isotopic composition of MTBE steadily changed from the source regions along the major contaminant plume (-26.4% to +40.0% (carbon); -73.1% to +60.3% (hydrogen)) indicating substantial biodegradation. Constant carbon isotopic signatures of TBA suggest the absence of TBA degradation at the site. Published carbon and hydrogen isotope fractionation data for biodegradation of MTBE under oxic and anoxic conditions, respectively, were examined and used to determine both the nature and the extent of in-situ biodegradation along the plume(s). The coupled evaluation of two-dimensional compound-specific isotope data explained both carbon and hydrogen fractionation data in a consistent way and indicate anaerobic biodegradation of MTBE along the entire plume. A novel scheme to reevaluate empiric isotopic enrichment factors (epsilon) in terms of theoretically based intrinsic carbon (12k/13k) and hydrogen (1k/2k) kinetic isotope effects (KIE) is presented. Carbon and hydrogen KIE values, calculated for different potential reaction mechanisms, imply that anaerobic biodegradation of MTBE follows a SN2-type reaction mechanism. Furthermore, our data suggest that additional removal process(es) such as evaporation contributed to the overall MTBE removal along the plume, a phenomenon that might be significant also for other field sites at tropic or subtropic climates with elevated groundwater temperatures (25 degrees C).

Bacteria, Anaerobic↗

PubMed2005

Requirement of aminoacyl-tRNA synthetases for gametogenesis and embryo development in Arabidopsis.

Aminoacyl-tRNA synthetases (AARSs) are required for translation in three different compartments of the plant cell: chloroplasts, mitochondria and the cytosol. Elimination of this basal function should result in lethality early in development. Phenotypes of individual mutants may vary considerably, depending on patterns of gene expression, functional redundancy, allele strength and protein localization. We describe here a reverse genetic screen of 50 insertion mutants disrupted in 21 of the 45 predicted AARSs in Arabidopsis. Our initial goal was to find additional EMB genes with a loss-of-function phenotype in the seed. Several different classes of knockouts were discovered, with defects in both gametogenesis and seed development. Three major trends were observed. Disruption of translation in chloroplasts often results in seed abortion at the transition stage of embryogenesis with minimal effects on gametophytes. Disruption of translation in mitochondria often results in ovule abortion before and immediately after fertilization. This early phenotype was frequently missed in prior screens for embryo-defective mutants. Knockout alleles of non-redundant cytosolic AARSs were in general not identified, consistent with the absolute requirement of cytosolic translation for development of male and female gametophytes. These results provide a framework for evaluating redundant functions of AARSs in Arabidopsis, a valuable data set of phenotypes resulting from multiple disruptions of a single basal process, and insights into which genes are required for both gametogenesis and embryo development and might therefore escape detection in screens for embryo-defective mutants.

Amino Acyl-tRNA Synthetases↗

PubMed2004

Identification of genes required for embryo development in Arabidopsis.

A long-term goal of Arabidopsis research is to define the minimal gene set needed to produce a viable plant with a normal phenotype under diverse conditions. This will require both forward and reverse genetics along with novel strategies to characterize multigene families and redundant biochemical pathways. Here we describe an initial dataset of 250 EMB genes required for normal embryo development in Arabidopsis. This represents the first large-scale dataset of essential genes in a flowering plant. When compared with 550 genes with other knockout phenotypes, EMB genes are enriched for basal cellular functions, deficient in transcription factors and signaling components, have fewer paralogs, and are more likely to have counterparts among essential genes of yeast (Saccharomyces cerevisiae) and worm (Caenorhabditis elegans). EMB genes also represent a valuable source of plant-specific proteins with unknown functions required for growth and development. Analyzing such unknowns is a central objective of genomics efforts worldwide. We focus here on 34 confirmed EMB genes with unknown functions, demonstrate that expression of these genes is not embryo-specific, validate a strategy for identifying interacting proteins through complementation with epitope-tagged proteins, and discuss the value of EMB genes in identifying novel proteins associated with important plant processes. Based on sequence comparison with essential genes in other model eukaryotes, we identify 244 candidate EMB genes without paralogs that represent promising targets for reverse genetics. These candidates should facilitate the recovery of additional genes required for seed development.

Animals↗

PubMed2003

Compound-specific stable isotope analysis of organic contaminants in natural environments: a critical review of the state of the art, prospects, and future challenges.

Compound-specific stable isotope analysis (CSIA) using gas chromatography-isotope ratio mass spectrometry (GC/IRMS) has developed into a mature analytical method in many application areas over the last decade. This is in particular true for carbon isotope analysis, whereas measurements of the other elements amenable to CSIA (hydrogen, nitrogen, oxygen) are much less routine. In environmental sciences, successful applications to date include (i) the allocation of contaminant sources on a local, regional, and global scale, (ii) the identification and quantification of (bio)transformation reactions on scales ranging from batch experiments to contaminated field sites, and (iii) the characterization of elementary reaction mechanisms that govern product formation. These three application areas are discussed in detail. The investigated spectrum of compounds comprises mainly n-alkanes, monoaromatics such as benzene and toluene, methyl tert-butyl ether (MTBE), polycyclic aromatic hydrocarbons (PAHs), and chlorinated hydrocarbons such as tetrachloromethane, trichloroethylene, and polychlorinated biphenyls (PCBs). Future research directions are primarily set by the state of the art in analytical instrumentation and method development. Approaches to utilize HPLC separation in CSIA, the enhancement of sensitivity of CSIA to allow field investigations in the microg L(-1) range, and the development of methods for CSIA of other elements are reviewed. Furthermore, an alternative scheme to evaluate isotope data is outlined that would enable estimates of position-specific kinetic isotope effects and, thus, allow one to extract mechanistic chemical and biochemical information.

Chromatography, High Pressure Liquid↗

PubMed2003

Compound-specific carbon isotope analysis of volatile organic compounds in the low-microgram per liter range.

Compound-specific carbon isotope analysis (CSIA) has become an important tool in biological, archeological, and geological studies as well as in forensics, food sciences, and organic chemistry. If sensitivity could be enhanced, CSIA would further have an improved potential for environmental applications such as, for example, in situ remediation studies to assess contaminated environments, identification of pollutant degradation pathways and kinetics, distinction between degradation/formation mechanisms, or, verification of contaminant sources. With this goal in mind, we have developed methods to determine delta13C values of commonly reported groundwater contaminants in low-microgram per liter concentrations. Several injection and preconcentration techniques were evaluated for this purpose, i.e., on-column injection, split/ splitless injection, solid-phase microextraction (SPME), and purge and trap (P&T) in combination with gas chromatography-isotope ratio mass spectrometry. The delta13C values of the target compounds were determined by liquid injections of the analytes dissolved in diethyl ether or, in the case of P&T and SPME, by extraction from water spiked with the analytes. P&T extraction was the most efficient preconcentration technique reaching method detection limits (MDLs) from 0.25 to 5.0 microg/L. These are the lowest MDLs reported so far for continuous-flow isotope ratio determinations, using a commercially available and fully automated system. Isotopic fractionation resulting from preconcentration and injection was investigated and quantified for the priority groundwater pollutants methyl tert-butyl ether (MTBE), chloroform, tetrachloromethane, chlorinated ethylenes, benzene, and toluene. The isotopic fractionations caused by the extraction techniques were small but highly reproducible and could therefore be corrected for. P&T was characterized by a higher reproducibility and smaller isotopic fractionations than SPME. Among the liquid injection techniques, cold on-column injection resulted in slightly better precision compared to split/splitless injection. However, the MDLs determined for liquid injections were 4-6 orders of magnitude higher (i.e., 9.5-2800 mg/L) than for P&T and SPME. Since both of the latter methods are solventless, a better chromatographic resolution was obtained than for the liquid injection techniques. The P&T and SPME methods described here are also applicable for CSIA of D/H ratios, which require 10-20 times higher analyte concentrations than 13C/12C analysis. Finally, the applicability of the described methods is demonstrated for pollutant concentrations of only 5-60 microg/L in environmental samples.

Journal Article↗

PubMed2003

Trihalomethane formation by chlorination of ammonium- and bromide-containing groundwater in water supplies of Hanoi, Vietnam.

The occurrence and the fate of trihalomethanes (THMs) in the water supply system of Hanoi City, Vietnam was investigated from 1998 to 2001. The chlorination efficiency, THM speciation, and, THM formation potential (THMFP) was determined in the water works and in tap water. With regard to THM formation, three types of groundwater resources were identified: (I) high bromide, (II) low bromide, and (III) high bromide combined with high ammonia and high dissolved organic carbon (DOC) concentrations. Under typical treatment conditions (total chlorine residual 0.5-0.8 mg/L), the total THM formation was always below WHO, EU, and USEPA drinking water standards and decreased in the order type I > type II > type III, although the THMFP was > 400 micrograms/L for type III water. The speciation showed > 80% of bromo-THMs in type I water due to the noticeable high bromide level (< or = 140 micrograms/L). In type II water, the bromo-THMs still accounted for some 40% although the bromide concentration is significantly lower (< or = 30 micrograms/L). In contrast, only traces of bromo-THMs were formed (approximately 5%) in type III water, despite bromide levels were high (< or = 240 micrograms/L). This observation could be explained by competition kinetics of chlorine reacting with ammonia and bromide. Based on chlorine exposure (CT) estimations, it was concluded that the current chlorination practice for type I and II waters is sufficient for > or = 2-log inactivation of Giardia lamblia cysts. However, in type III water the applied chlorine is masked as chloramine with a much lower disinfection efficiency. In addition to high levels of ammonia, type III groundwater is also contaminated by arsenic that is not satisfactory removed during treatment. N-nitrosodimethylamine, a potential carcinogen suspected to be formed during chloramination processes, was below the detection limit of 0.02 microgram/L in type III water.

Ammonia↗

PubMed2003

Role of capsular colanic acid in adhesion of uropathogenic Escherichia coli.

Urinary tract infections are the most common urologic disease in the United States and one of the most common bacterial infections of any organ system. Biofilms persist in the urinary tract and on catheter surfaces because biofilm microorganisms are resistant to host defense mechanisms and antibiotic therapy. The first step in the establishment of biofilm infections is bacterial adhesion; preventing bacterial adhesion represents a promising method of controlling biofilms. Evidence suggests that capsular polysaccharides play a role in adhesion and pathogenicity. This study focuses on the role of physiochemical and specific binding interactions during adhesion of colanic acid exopolysaccharide mutant strains. Bacterial adhesion was evaluated for isogenic uropathogenic Escherichia coli strains that differed in colanic acid expression. The atomic force microscope (AFM) was used to directly measure the reversible physiochemical and specific binding interactions between bacterial strains and various substrates as bacteria initially approach the interface. AFM results indicate that electrostatic interactions were not solely responsible for the repulsive forces between the colanic acid mutant strains and hydrophilic substrates. Moreover, hydrophobic interactions were not found to play a significant role in adhesion of the colanic acid mutant strains. Adhesion was also evaluated by parallel-plate flow cell studies in comparison to AFM force measurements to demonstrate that prolonged incubation times alter bacterial adhesion. Results from this study demonstrate that the capsular polysaccharide colanic acid does not enhance bacterial adhesion but rather blocks the establishment of specific binding as well as time-dependent interactions between uropathogenic E. coli and inert substrates.

Bacterial Adhesion↗

PubMed2002

Development and characterization of temperature-controlled microreactors for protein crystallization.

Physico-chemical properties of the crystallization of biological macromolecules are of particular interest for an efficient way to get high-quality crystals. Concept and realization of a novel temperature-controlled microreactor to study these parameters is revealed. The characterization of the device is focussed on the temperature distribution across the reaction chamber, its long-term stability and accuracy as well as the regeneration of the surface inside the chamber after contamination with a hydrophilic protein (rGFP). We conclude that a microreactor for in situ observation of a temperature-controlled batch crystallization is in place.

Chemical Phenomena↗

PubMed2002

Simultaneous determination of fuel oxygenates and BTEX using direct aqueous injection gas chromatography mass spectrometry (DAI-GC/MS).

A direct aqueous injection-gas chromatography/mass spectrometry (DAI-GC/MS) method for trace analysis of gasoline components in water is presented. The method allows for the simultaneous quantification of the following solutes: methyl tert-butyl ether (MTBE), its major degradation products (tert-butyl formate, tert-butyl alcohol (TBA), methyl acetate, and acetone), and possible substitutes of MTBE as an octane enhancer in gasoline (tert-amyl methyl ether, ethyl tert-butyl ether) as well as benzene, toluene, ethylbenzene, p-xylene, m-xylene, and o-xylene (BTEX). No enrichment or pretreatment steps are required, and sample volumes of only 50 microL are needed for analysis. The detection limits in two different matrixes (spiked lake water and contaminated groundwater) are < or = 2 microg/L for most analytes and < 0.2 microg/L for MTBE, benzene, and toluene. The accuracy of the DAI-GC/MS method was excellent as determined from comparison with headspace-GC/MS and purge-and-trap-GC/MS. The DAI-GC/MS method has been applied to various environmental studies, which demonstrated its versatility. The applications comprised both laboratory (MTBE degradation in water treatment, quantification of polar gasoline components) and field (MTBE degradation ata gasoline spill site) investigations.

Benzene↗

PubMed2002

Determination of liposome/water partition coefficients of organic acids and bases by solid-phase microextraction.

The extraction of two methylated anilines and three chlorinated phenols by solid-phase microextraction (SPME) fibers coated with polyacrylate was investigated as a function of pH. Only the neutral species of the acids and bases partitioned into the polymer. Extraction kinetics were accelerated for the hydrophobic phenols at pH values around their acidity constant. This is presumably due to a reconstitution of the neutral species in the unstirred aqueous layer adjacent to the polymer surface by the charged species through the fast acid-base equilibrium. Although the charged species is not taken up into the polymer, liposome/water distribution ratios could be measured up to a pH value, where 99% of the compounds were present as charged species. The partition coefficients of the neutral and charged species were extrapolated from the pH profiles of the liposome/water distribution ratios. The resulting values were slightly lower than those measured with equilibrium dialysis. The discrepancies are discussed with respect to differences in the experimental conditions and the possibility of matrix effects during SPME measurements.

Journal Article↗