Aerobic metabolic trichloroethene biodegradation under field-relevant conditions.

Chloroethenes belong to the most widely distributed groundwater contaminants. Since 2014, it has been known that trichloroethene (TCE) can be degraded aerobically and metabolically as growth substrate by a mixed bacterial enrichment culture (named SF culture). In this study, the degradation capabilities under a range of field-relevant conditions were investigated in fixed-bed reactors as well as in batch experiments. Aerobic metabolic TCE degradation was stable over the long term, with degradation optima at 22 °C and pH 7. Degradation of up to 400 µM TCE was observed. The longest starvation period after which degradation of TCE was regained was 112 days. The possible co-contaminants perchloroethene, trans-1,2-dichloroethene, and cis-1,2-dichloroethene did not inhibit TCE degradation, even though they were not degraded themselves. The presence of equimolar amounts of 1,1-dichloroethene and vinyl chloride inhibited TCE degradation. Experiments with groundwater from different chloroethene-contaminated field sites proved the potential of the SF culture for bioaugmentation. Thus, aerobic metabolic TCE degradation should be considered as a promising method for the bioremediation of field sites with TCE as the main contaminant.

Orientation dependence of microcirculation-induced diffusion signal in anisotropic tissues.

PURPOSE: To seek a better understanding of the effect of organized capillary flow on the MR diffusion-weighted signal. METHODS: A theoretical framework was proposed to describe the diffusion-weighted MR signal, which was then validated both numerically using a realistic model of capillary network and experimentally in an animal model of isolated perfused heart preparation with myocardial blood flow verified by means of direct arterial spin labeling measurements. RESULTS: Microcirculation in organized tissues gave rise to an MR signal that could be described as a combination of the bi-exponential behavior of conventional intravoxel incoherent motion (IVIM) theory and diffusion tensor imaging (DTI) -like anisotropy of the vascular signal, with the flow-related pseudo diffusivity represented as the linear algebraic product between the encoding directional unit vector and an appropriate tensor entity. Very good agreement between theoretical predictions and both numerical and experimental observations were found. CONCLUSION: These findings suggest that the DTI formalism of anisotropic spin motion can be incorporated into the classical IVIM theory to describe the MR signal arising from diffusion and microcirculation in organized tissues. Magn Reson Med 76:1252-1262, 2016. © 2015 Wiley Periodicals, Inc.

Aerobic biodegradation of cis-1,2-dichloroethene as sole carbon source: Stable carbon isotope fractionation and growth characteristics.

Cis-1,2-dichloroethene (cDCE) is a compound of concern at many chloroethene-contaminated sites, since it tends to accumulate during reductive dechlorination of the higher chlorinated ethenes. Stable carbon isotope fractionation during aerobic cDCE biodegradation was observed in groundwater microcosms under varying incubation conditions (room temperature/groundwater temperature; with/without inorganic nutrients), and resulted in an average stable carbon isotope enrichment factor of -15.2+/-0.5 per thousand. A new enrichment culture, obtained from groundwater microcosms, degraded cDCE concentrations up to 100mgL(-1), was active at temperatures between 4 and 23 degrees C, had a pH optimum of approximately 7, and could withstand prolonged periods (250d) of starvation. Microbial growth during degradation of cDCE as sole carbon and energy source was demonstrated by protein formation in mineral medium not containing any known auxiliary substrate. The obtained growth yield was 12.5+/-1.9g of proteinMol(-1) of cDCE, with a doubling time of 53+/-2h at 23 degrees C. Aerobic degradation of cDCE as sole carbon and energy source appears to be a promising biological process for site remediation.

Growth kinetics and stable carbon isotope fractionation during aerobic degradation of cis-1,2-dichloroethene and vinyl chloride.

Assessing changes in the isotopic signature of contaminants is a promising new tool to monitor microbial degradation processes. In this study, chloroethene degradation was proven by depletion of chloroethenes, formation of chloride, increase in protein content and stable carbon isotope fractionation. Aerobic degradation of vinyl chloride (VC) was found to proceed metabolically, with degradation rates of 0.48 and 0.29 d(-1); and growth yields of 9.7 and 6.4 g of protein/mol of VC at room and groundwater temperature, respectively. Cis-1,2-dichloroethene (cDCE) was degraded cometabolically under aerobic conditions when VC was provided as growth substrate. Aerobic degradation was associated with significant stable carbon isotope fractionation, with enrichment factors ranging from -5.4+/-0.4 per thousand for metabolic degradation of VC to -9.8+/-1.7 per thousand for cometabolic degradation of cDCE. Thus, it was demonstrated that stable carbon isotope fractionation is suitable for assessing aerobic chloroethene degradation, which can contribute significantly to site remediation.

Modulation of autocrine TNF-alpha-stimulated matrix metalloproteinase 9 (MMP-9) expression by mitogen-activated protein kinases in THP-1 monocytic cells.

Matrix metalloproteinase 9 (MMP-9) is implicated in various physiological processes by its ability to degrade the extracellular matrix (ECM) and process multiple regulatory proteins. Normally, MMP-9 expression is tightly controlled in cells. Sustained or enhanced MMP-9 secretion, however, has been demonstrated to contribute to the pathophysiology of numerous diseases, including arthritis and tumor progression, rendering this enzyme a major target for clinical interventions. Here we show that constitutive MMP-9 secretion was abrogated in THP-1 monocytic leukemia cells by addition of neutralizing antibodies against tumor necrosis factor alpha (TNF-alpha) or TNF receptor type 1 (TNF-R1), as well as by inhibition of TNF-alpha converting enzyme (TACE). This indicates that MMP-9 production in these cells is maintained by autocrine stimulation, with TNF-alpha acting via TNF-R1. To investigate the intracellular signaling routes involved in MMP-9 gene transcription, cells were treated with different inhibitors of major mitogen-activated protein kinase (MAPK) pathways. Interruption of the extracellular signal-regulated kinase pathway 1/2 (ERK1/2) using PD98059 significantly downregulated constitutive MMP-9 release. In contrast, blockage of p38 kinase activity by addition of SB203580 or SB202190, as well as inhibition of c-Jun N-terminal kinase (JNK) using L-JNK-I1, clearly augmented MMP-9 expression and secretion by an upregulation of ERK1/2 phosphorylation. Moreover, exogenously added TNF-alpha augmented MMP-9 synthesis and secretion in THP-1 cells via enhancement of ERK1/2 activity. Taken together, our results indicate that ERK1/2 activity plays a pivotal role in TNF-alpha-induced MMP-9 production and demonstrate its negative modulation by p38 and JNK activity. These findings suggest ERK1/2 rather than p38 and JNK as a reasonable target to specifically block MMP-9 expression using MAPK inhibitors in therapeutic applications.

The calpastatin-derived calpain inhibitor CP1B reduces mRNA expression of matrix metalloproteinase-2 and -9 and invasion by leukemic THP-1 cells.

The ubiquitous proteases mu- and m-calpain are Ca(2+)-dependent cysteine endopeptidases. Besides involvement in a variety of physio(patho)logical processes, recent studies suggest a pivotal role of calpains in differentiation of hematopoietic cells and tumor cell invasion. However, the precise actions of calpains and their endogenous inhibitor, calpastatin, in these processes are only partially understood. Here we have studied the role of the calpain/calpastatin system in the invasion of leukemic cells under basal and differentiation-stimulating conditions. To further differentiate the human leukaemic cell line THP-1 (monocytic), the cells were treated for 24 hours with the differentiation-stimulating reagents phorbol 12-myristate 13-acetate (PMA) and dimethyl sulfoxide (DMSO). Macrophage- and granulocyte-like differentiation was confirmed by induction of vimentin expression as well as by microscopic and fluorescence-assisted cytometric analysis. Extracellular matrix (ECM) invasion of both the basal and differentiation-stimulated cells in a Matrigel assay was inhibited by pre-incubation of the cells with the specific calpain inhibitor CP1B for 24 hours. Inhibition of invasiveness correlated with decreased mRNA expression and secretion of the matrix metalloproteinases MMP-2 and MMP-9. In contrast, addition of CP1B only during the invasion process did neither influence transmigration nor MMP release. This is the first report showing that the calpain/calpastatin system mediates MMP-mRNA expression of the leukemic THP-1 cells and as a consequence their invasiveness.