A randomized clinical trial of cognitive behavioral therapy and interpersonal psychotherapy for panic disorder with agoraphobia.

BACKGROUND: Interpersonal psychotherapy (IPT) seems to be as effective as cognitive behavioral therapy (CBT) in the treatment of major depression. Because the onset of panic attacks is often related to increased interpersonal life stress, IPT has the potential to also treat panic disorder. To date, a preliminary open trial yielded promising results but there have been no randomized controlled trials directly comparing CBT and IPT for panic disorder. METHOD: This study aimed to directly compare the effects of CBT versus IPT for the treatment of panic disorder with agoraphobia. Ninety-one adult patients with a primary diagnosis of DSM-III or DSM-IV panic disorder with agoraphobia were randomized. Primary outcomes were panic attack frequency and an idiosyncratic behavioral test. Secondary outcomes were panic and agoraphobia severity, panic-related cognitions, interpersonal functioning and general psychopathology. Measures were taken at 0, 3 and 4 months (baseline, end of treatment and follow-up). RESULTS: Intention-to-treat (ITT) analyses on the primary outcomes indicated superior effects for CBT in treating panic disorder with agoraphobia. Per-protocol analyses emphasized the differences between treatments and yielded larger effect sizes. Reductions in the secondary outcomes were equal for both treatments, except for agoraphobic complaints and behavior and the credibility ratings of negative interpretations of bodily sensations, all of which decreased more in CBT. CONCLUSIONS: CBT is the preferred treatment for panic disorder with agoraphobia compared to IPT. Mechanisms of change should be investigated further, along with long-term outcomes.

Flux-based risk management strategy of groundwater pollutions: the CMF approach.

A site- and receptor-specific risk management strategy for groundwater pollution based on the measurement of contaminant mass flux is proposed. The approach is useful and compatible with the demands formulated in the European Water Framework Directive, its Groundwater Daughter Directive and the regulations applicable in the EU member states. The proposed CMF method focuses on the following: (1) capture zones, (2) the location of control planes, (3) the definition of the maximum allowed contaminant mass discharge and (4) contaminant mass flux measurements. For every control plane, such a maximum allowed contaminant mass discharge is derived and is crucial for the receptor risk management strategy. The method is demonstrated for a large area of groundwater pollution present in the industrial area of Vilvoorde-Machelen located in Flanders, Belgium.

Microbial processes as key drivers for metal (im)mobilization along a redox gradient in the saturated zone.

Two sites representing different aquifer types, i.e., Dommel (sandy) and Flémalle (gravelly loam) along the Meuse River, have been selected to conduct microcosm experiments. Various conditions ranging from aerobic over nitrate- to sulphate reducing were imposed. For the sandy aquifer, nitrate reducing conditions predominated, which specifically in the presence of a carbon source led to pH increases and enhanced Zn removal. For the calcareous gravelly loam, sulphate reduction was dominant resulting in immobilization of both Zn and Cd. For both aquifer types and almost all redox conditions, higher arsenic concentrations were measured in the groundwater. Analyses of different specific microbial populations by polymerase chain reaction (PCR) revealed the dominance of denitrifiers for the Dommel site, while sulfate reducing bacteria (SRB) were the prevailing population for all redox conditions in the Flémalle samples.

Plasmids for heavy metal resistance in Alcaligenes eutrophus CH34: mechanisms and applications.

Alcaligenes eutrophus CH34 is the main representative of a group of strongly related strains (mostly facultative chemolithotrophs) that are well adapted to environments containing high levels of heavy metals. It harbors the megaplasmids pMOL28 and pMOL30 which carry resistance determinants to Co2+, Ni2+, CrO(4)2-, Hg2+, Tl+, Cd2+, Cu2+ and Zn2+. Among the best characterized determinants are the cnr operon (resistance to Co, Ni) on pMOL28 and the czc operon on pMOL30 (resistance to Co, Cd and Zn). Although the two systems reveal a significant degree of amino acid similarity in the structural genes, the regulation of the operons is different. The resistance mechanism in both cases is based on efflux. The efflux mechanism leads to a pH increase outside of the cytoplasmic membrane. Metals are sequestered from the external medium through the bioprecipitation of metal carbonates formed in the saturated zone around the cell. This latter phenomenon can be exploited in bioreactors designed to remove metals from effluents. The bacteria are immobilized on composite membranes in a continuous tubular membrane reactor (CTMR). The effluent continuously circulates through the intertubular space, while the external surface of the tubes is in contact with the growth medium. Metal crystals are eventually removed by the effluent stream and collected on a glass bead column. The system has been applied to effluents containing Cd, Zn, Co, Ni and Cu. By introducing catabolic plasmids involved in the aerobic degradation of PCBs and 2,4-D into metal-resistant A. eutrophus strains, the application range was widened to include effluents polluted with both organic and inorganic substances. Biosensors have been developed which are based on the fusion of genes induced by metals to a reporter system, the lux operon of Vibrio fischeri. Bacterial luciferases produce light through the oxidation of fatty aldehydes. The gene fusions are useful both for the study of regulatory genes and for the determination of heavy metal concentrations in the environment.

Groundwater Flow Field Distortion by Monitoring Wells and Passive Flux Meters.

Due to differences in hydraulic conductivity and effects of well construction geometry, groundwater lateral flow through a monitoring well typically differs from groundwater flow in the surrounding aquifer. These differences must be well understood in order to apply passive measuring techniques, such as passive flux meters (PFMs) used for the measurement of groundwater and contaminant mass fluxes. To understand these differences, lab flow tank experiments were performed to evaluate the influences of the well screen, the surrounding filter pack and the presence of a PFM on the natural groundwater flux through a monitoring well. The results were compared with analytical calculations of flow field distortion based on the potential theory of Drost et al. (1968). Measured well flow field distortion factors were found to be lower than calculated flow field distortion factors, while measured PFM flow field distortion factors were comparable to the calculated ones. However, this latter is not the case for all conditions. The slotted geometry of the well screen seems to make a correct analytical calculation challenging for conditions where flow field deviation occurs, because the potential theory assumes a uniform flow field. Finally, plots of the functional relationships of the distortion of the flow field with the hydraulic conductivities of the filter screen, surrounding filter pack and corresponding radii make it possible to design well construction to optimally function during PFM applications.

A transcriptional luxAB reporter fusion responding to fluorene in Sphingomonas sp. LB126 and its initial characterisation for whole-cell bioreporter purposes.

The promoter probe mini-Tn5-luxAB-tet was used to create a luxAB transcriptional fusion responding to fluorene in the fluorene utilising bacterium Sphingomonas sp. LB126. The mutant strain, named L-132, was impaired in fluorene utilisation and strongly emitted light upon addition of fluorene to the growth medium. L-132 was initially characterised and examined for its potential use as a whole-cell biosensor in the perspective of quantifying fluorene in environmental samples. Activity of the reporter gene as a response to fluorene was detectable after 30 min and was optimal after 4 h. A linear response to fluorene concentrations within the water solubility range was achieved, with a detection limit of 200 microg per litre. Besides fluorene, L-132 weakly responded to the polycyclic aromatic hydrocarbons phenanthrene and dibenzothiophene, whereas strong responses were obtained with 9-fluorenone, 9-hydroxyfluorene, phthalic acid and protocatechuic acid. The latter four compounds are metabolites formed in course of fluorene degradation, which suggested that a fluorene metabolite rather than fluorene itself was the true inducer of the luxAB fusion in L-132.

Heavy metals bioremediation of soil.

Historical emissions of old nonferrous factories lead to large geographical areas of metals-contaminated sites. At least 50 sites in Europe are contaminated with metals like Zn, Cd, Cu, and Pb. Several methods, based on granular differentiation, were developed to reduce the metals content. However, the obtained cleaned soil is just sand. Methods based on chemical leaching or extraction or on electrochemistry do release a soil without any salts and with an increased bioavailability of the remaining metals content. In this review a method is presented for the treatment of sandy soil contaminated with heavy metals. The system is based on the metal solubilization on biocyrstallization capacity of Alcaligenes eutrophus CH34. The bacterium can solubilize the metals (or increase their bioavailability) via the production of siderophores and adsorb the metals in their biomass on metal-induced outer membrane proteins and by bioprecipitation. After the addition of CH34 to a soil slurry, the metals move toward the biomass. As the bacterium tends to float quite easily, the biomass is separated from the water via a flocculation process. The Cd concentration in sandy soils could be reduced from 21 mg Cd/kg to 3.3 mg Cd/kg. At the same time, Zn was reduced from 1070 mg Zn/kg to 172 mg Zn/kg. The lead concentration went down from 459 mg Pb/kg to 74 mg Pb/kg. With the aid of biosensors, a complete decrease in bioavailability of the metals was measured.

Development and validation of a liquid chromatographic-tandem mass spectrometric method for the determination of galantamine in human heparinised plasma.

Galantamine is an acetylcholinesterase inhibitor, recently approved for the treatment of mild-to-moderate Alzheimer's disease. To allow a higher throughput of samples, a new bioanalytical method for the determination of galantamine in human plasma was developed and validated. A stable isotope labelled internal standard was used. Sample preparation consisted of a simple one-step liquid-liquid extraction with toluene. The extracts were analysed with positive ion TurboIonspray tandem mass spectrometry (LC-MS-MS). The method was validated in the 1-500-ng/ml range. The accuracy, precision, selectivity, lower limit of quantification, upper limit of quantification, linearity and extraction recovery were evaluated, as well as the stability of the compound in plasma, blood, methanol and 2% BSA solutions under different conditions. The method proved very rugged during the analysis of large numbers of samples from clinical trials.

Use of DNA probes and plasmid capture in a search for new interesting environmental genes.

Adaptation to a stressed environment leads to organisms bearing DNA, encoding defense mechanisms. These mechanisms can be heavy metal resistance, catabolism of organic xenobiotics or stress reactions. Genes responsible for these mechanisms can be used for monitoring changing environments and therefore it can be important to store such bacteria in a bank. DNA-probing will be presented by the use of DNA fragments (of Alcaligenes eutrophus) coding for heavy metal resistance or xenobiotic degradation. Some strains do not grow on petri dishes and accordingly cannot be isolated from soils. In order to isolate plasmids from such strains, coding for heavy metal resistances or xenobiotic degradations, an exogenous plasmid isolation method was developed. In this method, the endogenous population is conjugated with Pseudomonas or Alcaligenes strains bearing a retrotransfer plasmid like RP4. In that way new plasmids from various sources including non-culturable strains could be obtained. With these methods, a large number of specimens adapted to stressed situations can be isolated or constructed (in the case of the exogenous plasmid isolation method). They form a source of interesting genetic material that can be used to restore polluted areas in natural areas, if necessary with the aid of genetic engineering (in vitro or in vivo techniques). Full knowledge of such bacteria and their resistance mechanisms or degradation pathways, can lead to new constructions able to attack recalcitrant mixtures of different organics and to resist heavy metals.

Effect of drying on bioremediation bacteria properties.

Bioremediation bacteria with drought-resistance characteristics were selected and compared to a collection of 10 strains selected only for their bioremediation properties. Twenty-six strains were selected from dried diesel-polluted soil, and they exhibit a better level of survival during drying, compared to collection bioremediation strains (two orders of magnitude difference). The lyophilization process does not affect the strains' ability to grow on xenobiotic compound when measured immediately after drying. However, collection bioremediation strains selected only for their bioremediation properties lose up to 80% of their properties when stored at 25 degrees C for 15 d, but the strains selected for their drought resistance lose their properties to a lesser extent during the same period. The maximal growth rate and the rate of xenobiotic degradation of the still-active cells are not affected by the drying process.

Thermal degradation of cloudy apple juice phenolic constituents.

Although conventional thermal processing is still the most commonly used preservation technique in cloudy apple juice production, detailed knowledge on phenolic compound degradation during thermal treatment is still limited. To evaluate the extent of thermal degradation as a function of time and temperature, apple juice samples were isothermally treated during 7,200s over a temperature range of 80-145 °C. An untargeted metabolomics approach based on liquid chromatography-high resolution mass spectrometry was developed and applied with the aim to find out the most heat labile phenolic constituents in cloudy apple juice. By the use of a high resolution mass spectrometer, the high degree of in-source fragmentation, the quality of deconvolution and the employed custom-made database, it was possible to achieve a high degree of structural elucidation for the thermolabile phenolic constituents. Procyanidin subclass representatives were discovered as the most heat labile phenolic compounds of cloudy apple juice.

An improved mass spectrometric method for identification and quantification of phenolic compounds in apple fruits.

Thirty-nine phenolic compounds were analysed using ultra high performance liquid chromatography (UHPLC) coupled with diode array and accurate mass spectrometry detection using electrospray ionisation (DAD/ESI-am-MS). Instrumental parameters such as scan speed, resolution, and mass accuracy were optimised to establish accurate mass measurements. The method was fully validated in terms of model deviation (r(2)>0.9990), range (typically 10-3500 ngg(-1)), intra/inter-day precision (<6% and <8%, respectively) and accuracy (typically 100 ± 10%). The mass accuracy of each selected phenolic compound was below 1.5 ppm. The results confirmed that the UHPLC-DAD/ESI-am-MS method developed here was convenient and reliable for the determination of phenolic compounds in apple extracts.

Field demonstration and evaluation of the Passive Flux Meter on a CAH groundwater plume.

This study comprises the first application of the Passive Flux Meter (PFM) for the measurement of chlorinated aliphatic hydrocarbon (CAH) mass fluxes and Darcy water fluxes in groundwater at a European field site. The PFM was originally developed and applied to measurements near source zones. The focus of the PFM is extended from near source to plume zones. For this purpose, 48 PFMs of 1.4 m length were constructed and installed in eight different monitoring wells in the source and plume zone of a CAH-contaminated field site located in France. The PFMs were retrieved, sampled, and analyzed after 3 to 11 weeks of exposure time, depending on the expected contaminant flux. PFM evaluation criteria include analytical, technical, and practical aspects as well as conditions and applicability. PFM flux data were compared with so-called traditional soil and groundwater concentration data obtained using active sampling methods. The PFMs deliver reasonable results for source as well as plume zones. The limiting factor in the PFM applicability is the exposure time together with the groundwater flux. Measured groundwater velocities at the field site range from 2 to 41 cm/day. Measured contaminant flux data raise up to 13 g/m(2)/day for perchloroethylene in the plume zone. Calculated PFM flux averaged concentration data and traditional concentration data were of similar magnitude for most wells. However, both datasets need to be compared with reservation because of the different sampling nature and time. Two important issues are the PFM tracer loss during installation/extraction and the deviation of the groundwater flow field when passing the monitoring well and PFM. The demonstration of the PFM at a CAH-contaminated field site in Europe confirmed the efficiency of the flux measurement technique for source as well as plume zones. The PFM can be applied without concerns in monitoring wells with European standards. The acquired flux data are of great value for the purpose of site characterization and mass discharge modeling, and can be used in combination with traditional soil and groundwater sampling methods.

Selective carboxylate production by controlling hydrogen, carbon dioxide and substrate concentrations in mixed culture fermentation.

This research demonstrated the selective production of n-butyrate from mixed culture by applying 2 bar carbon dioxide into the headspace of batch fermenters or by increasing the initial substrate concentration. The effect of increasing initial substrate concentration was investigated at 8, 13.5 and 23 g COD/L with potato processing waste stream. Within 1 week of incubation, n-butyrate fraction selectively increased up to 83% by applying 2 bar hydrogen or 78% by applying carbon dioxide into the headspace whereas it was only 59% in the control reactor. Although the fraction of n-butyrate was elevated, the concentration remained lower than in the control. Both the highest concentration and fraction of n-butyrate were observed under the highest initial substrate concentration without headspace addition. The concentration was 10 g COD/L with 73% fraction. The operational conditions obtained from batch experiments for selective n-butyrate production were validated in a continuous process.

Effect of hydrogen and carbon dioxide on carboxylic acids patterns in mixed culture fermentation.

This study investigated the carboxylate spectrum from mixed culture fermentation of three organic waste streams after supplying 2 bar hydrogen and carbon dioxide or a mixture of these two gases to the headspace. Under any modified headspace, propionate production was ceased and butyrate, caproate and the total carboxylate concentrations were higher than in the reactors with N(2) headspace (control). Production of one major compound was achieved under hydrogen and carbon dioxide mixed headspace after 4 weeks of incubation. Both the highest acetate concentration (17.4 g COD/l) and the highest fraction (87%) were observed in reactors with mixed hydrogen and carbon dioxide headspace independent of the substrate used. In the control reactor, acetate made up maximum 67% of the total products. For other products, the highest concentration and fraction were seldom observed together. Selective butyrate production reaching a 75% fraction was found under the carbon dioxide headspace on the carbohydrate rich waste.

Potential use of the plant antioxidant network for environmental exposure assessment of heavy metals in soils.

In recent years, awareness has risen that the total soil content of pollutants by itself does not suffice to fully assess the potential ecotoxicological risks involved. Chemical analysis will require to be complemented with biological assays in a multidisciplinary approach towards site specific ecological risk assessment (SS-ERA). This paper evaluates the potential use of the plants' antioxidant response to metal-induced oxidative stress to provide a sensitive biological assay in SS-ERA. To this end, plants of Phaseolus vulgaris were grown for two weeks on 15 soils varying in contamination level. Morphological parameters and enzymatic plant responses were measured upon harvest. Foliar concentrations of the (heavy) metals Al, Cu, Cd, Cr, Fe, Mn, Ni, Pb, Zn were also determined. Metal mobility in the soil was further assessed by determining soil solution and NH4OAc extractable levels. In general more significant correlations were observed between plant responses and foliar metal concentrations or exchangeable/soluble levels than between plant responses and the total soil content. The study demonstrates the potential use of the plants' antioxidant defence mechanisms to assess substrate phytotoxicity for application in SS-ERA protocols. However, the system, based on calculation of a soil Phytotoxicity Index (PI), will require adaptation and fine-tuning to meet the specific needs for this type of environmental monitoring. Large variation was observed in phytotoxicity classification based on the various test parameters. The thresholds for classification of the various morphological and enzymatic response parameters may require adaptation according to parameter stress sensitivity in order to decrease the observed variation. The use of partial PI's (leaves and roots separately) may in addition increase the sensitivity of the system since some metals show specific effects in one of both organs only. Loss of biological functionality of enzymes, as was observed for ICDH in one of the more strongly contaminated soils, may also be recognized as an additional stress symptom when assigning phytotoxicity classification, whereas the current system only considers increasing enzymatic capacities. Other easily distinguishable parameters, which could be added to the current indexation are: failure to germinate and the incapacity to develop roots in the toxic substrate. Additional research will be required to determine the possible application range of soil properties for this biological assay and to further improve its performance in SS-ERA.

DNA probe-mediated detection of resistant bacteria from soils highly polluted by heavy metals.

Alcaligenes eutrophus CH34 DNA fragments encoding resistance to Cd, Co, Zn (czc), or Hg (merA) were cloned and used as probes in colony hybridization procedures with bacteria isolated from polluted environments such as a zinc factory area (desertified because of the toxic effects of zinc contamination) and from sediments from factories of nonferrous metallurgy in Belgium and mine areas in Zaire. From the different soil samples, strains could be isolated and hybridized with the czc probe (resistance to Cd, Co, and Zn from plasmid pMOL30). Percentages of CFU isolated on nonselective plates which hybridized with the czc and the mercury resistance probes were, respectively, 25 and 0% for the zinc desert, 15 to 20 and 10 to 20% for the two Belgian factories, and 40 and 40% for the Likasi mine area. Most of these strains also carried two large plasmids of about the same size as those of A. eutrophus CH34 and shared many phenotypic traits with this strain. These findings indicated a certain correlation between the heavy-metal content in contaminated soils and the presence of heavy-metal-resistant megaplasmid-bearing A. eutrophus strains.