A sensitized ENU mutagenesis screen for dominant genetic modifiers of thrombosis in the factor V Leiden mouse

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106086/1/jth05193.pd

Deficiency of plasminogen activator inhibitor‐2 results in accelerated tumor growth

BackgroundUpregulation of the plasminogen activation system, including urokinase plasminogen activator (uPA), has been observed in many malignancies, suggesting that co‐opting the PA system is a common method by which tumor cells accomplish extracellular matrix proteolysis. PAI‐2, a serine protease inhibitor, produced from the SERPINB2 gene, inhibits circulating and extracellular matrix‐tethered uPA. Decreased SERPINB2 expression has been associated with increased tumor invasiveness and metastasis for several types of cancer. PAI‐2 deficiency has not been reported in humans and PAI‐2‐deficient (SerpinB2−/−) mice exhibit no apparent abnormalities.ObjectivesWe investigated the role of PAI‐2 deficiency on tumor growth and metastasis.MethodsTo explore the long‐term impact of PAI‐2 deficiency, a cohort of SerpinB2−/− mice were aged to >18 months, with spontaneous malignancies observed in 4/9 animals, all of apparently vascular origin. To further investigate the role of PAI‐2 deficiency in malignancy, SerpinB2−/− and wild‐type control mice were injected with either B16 melanoma or Lewis lung carcinoma tumor cells, with markedly accelerated tumor growth observed in SerpinB2−/− mice for both cell lines. To determine the relative contributions of PAI‐2 from hematopoietic or nonhematopoietically derived sources, bone marrow transplants between wild‐type C57BL/6J and SerpinB2−/− mice were performed.Results and ConclusionsOur results suggest that PAI‐2 deficiency increases susceptibility to spontaneous tumorigenesis in the mouse, and demonstrate that SerpinB2 expression derived from a nonhematopoietic compartment is a key host factor in the regulation of tumor growth in both the B16 melanoma and Lewis lung carcinoma models.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/163438/2/jth15054_am.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/163438/1/jth15054.pd

The factor V Leiden mutation results in the in vivo loss of a critical FV anticoagulant function

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106106/1/jth05190.pd

Treatment of experimental adjuvant arthritis with a novel folate receptor-targeted folic acid-aminopterin conjugate

Introduction Folate receptor (FR)-expressing macrophages have been shown to accumulate at sites of inflammation, where they promote development of inflammatory symptoms. To target such a macrophage population, we designed and evaluated the biologic activity of EC0746, a novel folic acid conjugate of the highly potent antifolate, aminopterin. Methods Using a FR-positive subclone of murine macrophage-derived RAW264.7 cells and rat thioglycollate-elicited macrophages, we studied the effect of EC0746 on dihydrofolate reductase activity, cell proliferation, and cellular response towards bacterial lipopolysaccharide as well as IFNγ activation. The EC0746 anti-inflammatory activity, pharmacokinetics, and toxicity were also evaluated in normal rats or in rats with adjuvant-induced arthritis; that is, a FR-positive macrophage model that closely resembles rheumatoid arthritis in humans. Results EC0746 suppresses the proliferation of RAW264.7 cells and prevents the ability of nonproliferating rat macrophages to respond to inflammatory stimuli. In the macrophage-rich rat arthritis model, brief treatment with subcutaneously administered EC0746 is shown to mediate an FR-specific anti-inflammatory response that is more potent than either orally administered methotrexate or subcutaneously delivered etanercept. More importantly, EC0746 therapy is also shown to be ~40-fold less toxic than unmodified aminopterin, with fewer bone marrow and gastrointestinal problems. Conclusions EC0746 is the first high FR-binding dihydrofolate reductase inhibitor that demonstrates FR-specific anti-inflammatory activities both in vitro and in vivo. Our data reveal that a relatively toxic anti-inflammatory drug, such as aminopterin, can be targeted with folic acid to inflammatory macrophages and thereby relieve inflammatory symptoms with greatly reduced toxicity

Temporal evolution of shallow marine diagenetic environments: Insights from carbonate concretions

Early diagenesis of marine organic matter dramatically impacts Earth's surface chemistry by changing the burial potential of carbon and promoting the formation of authigenic mineral phases including carbonate concretions. Marine sediment-hosted carbonate concretions tend to form as a result of microbial anaerobic diagenetic reactions that degrade organic matter and methane, some of which require an external oxidant. Thus, temporal changes in the oxidation state of Earth's oceans may impart a first-order control on concretion authigenesis mechanisms through time. Statistically significant variability in concretion carbonate carbon isotope compositions indicates changes in shallow marine sediment diagenesis associated with Earth's evolving redox landscape. This variability manifests itself as an expansion in carbon isotope composition range broadly characterized by an increase in maximum and decrease in minimum isotope values through time. Reaction transport modelling helps to constrain the potential impacts of shifting redox chemistry and highlights the importance of organic carbon delivery to the seafloor, marine sulfate concentrations, methane production and external methane influx. The first appearance of conclusively anaerobic oxidation of methane-derived concretions occurs in the Carboniferous and coincides with a Paleozoic rise in marine sulfate. The muted variability recognized in older concretions (and in particular for Precambrian concretions) likely reflects impacts of a smaller marine sulfate reservoir and perhaps elevated marine dissolved inorganic carbon concentrations. Causes of the increase in carbon isotope maximum values through time are more confounding, but may be related to isotopic equilibration of dissolved inorganic carbon with externally derived methane. Ultimately the concretion isotope record in part reflects changes in organic matter availability and marine oxidation state, highlighting connections with the subsurface biosphere and diagenesis throughout geologic time

Ambient BTEX exposure and mid-pregnancy inflammatory biomarkers in pregnant African American women

Air pollution is associated with preterm birth (PTB), potentially via inflammation. We recently showed the mixture benzene, toluene, ethylbenzene, and xylene (BTEX) is associated with PTB. We examined if ambient BTEX exposure is associated with mid-pregnancy inflammation in a sample of 140 African-American women residing in Detroit, Michigan. The Geospatial Determinants of Health Outcomes Consortium study collected outdoor air pollution measurements in Detroit; these data were coupled with Michigan Air Sampling Network measurements to develop monthly BTEX concentration estimates at a spatial density of 300 m(2). First trimester and mid-pregnancy BTEX exposure estimates were assigned to maternal address. Mid-pregnancy (mean 21.3 ± 3.7 weeks gestation) inflammatory biomarkers (high-sensitivity C-reactive protein, interleukin [IL]-6, IL-10, IL-1β, and tumor necrosis factor-α) were measured with enzyme immunoassays. After covariate adjustment, for every 1-unit increase in first trimester BTEX, there was an expected mean increase in log-transformed IL-1β of 0.05 ± 0.02 units (P = 0.014) and an expected mean increase in log-transformed tumor necrosis factor-α of 0.07 ± 0.02 units (P = 0.006). Similarly, for every 1-unit increase in mid-pregnancy BTEX, there was a mean increase in log IL-1β of 0.06 ± 0.03 units (P = 0.027). There was no association of either first trimester or mid-pregnancy BTEX with high-sensitivity C-reactive protein, IL-10, or IL-6 (all P \u3e 0.05). Ambient BTEX exposure is associated with inflammation in mid-pregnancy in African-American women. Future studies examining if inflammation mediates associations between BTEX exposure and PTB are needed

Drone-based Water Sampling and Characterization of Three Freshwater Harmful Algal Blooms in the United States

Freshwater harmful algal blooms (HABs), caused mostly by toxic cyanobacteria, produce a range of cyanotoxins that threaten the health of humans and domestic animals. Climate conditions and anthropogenic influences such as agricultural run-off can alter the onset and intensity of HABs. Little is known about the distribution and spread of freshwater HABs. Current sampling protocols in some lakes involve teams of researchers that collect samples by hand from a boat and/or from the shoreline. Water samples can be collected from the surface, from discrete-depth collections, and/or from depth-integrated intervals. These collections are often restricted to certain months of the year, and generally are only performed at a limited number of collection sites. In lakes with active HABs, surface samples are generally sufficient for HAB water quality assessments. We used a unique DrOne Water Sampling SystEm (DOWSE) to collect water samples from the surface of three different HABs in Ohio (Grand Lake St Marys, GLSM and Lake Erie) and Virginia (Lake Anna), United States in 2019. The DOWSE consisted of a 3D-printed sampling device tethered to a drone (uncrewed aerial system, or UAS), and was used to collect surface water samples at different distances (10–100 m) from the shore or from an anchored boat. One hundred and eighty water samples (40 at GLSM, 20 at Lake Erie, and 120 at Lake Anna) were collected and analyzed from 18 drone flights. Our methods included testing for cyanotoxins, phycocyanin, and nutrients from surface water samples. Mean concentrations of microcystins (MCs) in drone water samples were 15.00, 1.92, and 0.02 ppb for GLSM, Lake Erie, and Lake Anna, respectively. Lake Anna had low levels of anatoxin in nearly all (111/120) of the drone water samples. Mean concentrations of phycocyanin in drone water samples were 687, 38, and 62 ppb for GLSM, Lake Erie, and Lake Anna, respectively. High levels of total phosphorus were observed in the drone water samples from GLSM (mean of 0.34 mg/L) and Lake Erie (mean of 0.12 mg/L). Lake Anna had the highest variability of total phosphorus with concentrations that ranged from 0.01 mg/L to 0.21 mg/L, with a mean of 0.06 mg/L. Nitrate levels varied greatly across sites, inverse with bloom biomass, ranging from below detection to 3.64 mg/L, with highest mean values in Lake Erie followed by GLSM and Lake Anna, respectively. Drones offer a rapid, targeted collection of water samples from virtually anywhere on a lake with an active HAB without the need for a boat which can disturb the surrounding water. Drones are, however, limited in their ability to operate during inclement weather such as rain and heavy winds. Collectively, our results highlight numerous opportunities for drone-based water sampling technologies to track, predict, and respond to HABs in the future

Contamination of Chalk groundwater by chlorinated solvents : a case study of deep penetration by non-aqueous phase liquids

The transport behaviour of chlorinated solvents, both in the aqueous phase and as a dense non-aqueous phase liquid (DNAPL), in fissured microporous aquifers is reviewed. The presence of DNAPL in aquifers is especially serious as it is likely to be the main subsurface source of contamination and, given the slow rates of dissolution in groundwater, may persist for decades. However, the identification and quantification of DNAPLs in fractured aquifers present many practical problems and are often not achievable. A case study of a Chalk site which had been contaminated by chlorinated solvents demonstrated that the use of a range of techniques, including depth profiling of solvent porewater concentrations in cored boreholes, can provide clear evidence for the presence of DNAPL at depth, although DNAPL was not itself observed. Theoretical considerations and field observations confirmed that DNAPL movement is via fractures rather than through the microporous matrix