Microbial Community Functional Potential and Composition Are Shaped by Hydrologic Connectivity in Riverine Floodplain Soils.

Riverine floodplains are ecologically and economically valuable ecosystems that are heavily threatened by anthropogenic stressors. Microbial communities in floodplain soils mediate critical biogeochemical processes, yet we understand little about the relationship between these communities and variation in hydrologic connectivity related to land management or topography. Here, we present metagenomic evidence that differences among microbial communities in three floodplain soils correspond to a long-term gradient of hydrologic connectivity. Specifically, all strictly anaerobic taxa and metabolic pathways were positively associated with increased hydrologic connectivity and flooding frequency. In contrast, most aerobic taxa and all strictly aerobic pathways were negatively related to hydrologic connectivity and flooding frequency. Furthermore, the genetic potential to metabolize organic compounds tended to decrease as hydrologic connectivity increased, which may reflect either the observed concomitant decline of soil organic matter or the parallel increase in both anaerobic taxa and pathways. A decline in soil N, accompanied by an increased genetic potential for oligotrophic N acquisition subsystems, suggests that soil nutrients also shape microbial communities in these soils. We conclude that differences among floodplain soil microbial communities can be conceptualized along a gradient of hydrologic connectivity. Additionally, we show that these differences are likely due to connectivity-related variation in flooding frequency, soil organic matter, and soil N. Our findings are particularly relevant to the restoration and management of microbially mediated biogeochemical processes in riverine floodplain wetlands.

A role for glutathione, independent of oxidative stress, in the developmental toxicity of methanol.

Oxidative stress and reactive oxygen species (ROS) have been implicated in the teratogenicity of methanol (MeOH) in rodents, both in vivo and in embryo culture. We explored the ROS hypothesis further in vivo in pregnant C57BL/6J mice. Following maternal treatment with a teratogenic dose of MeOH, 4 g/kg via intraperitoneal (ip) injection on gestational day (GD) 12, there was no increase 6h later in embryonic ROS formation, measured by 2',7'-dichlorodihydrofluorescin diacetate (DCFH-DA) fluorescence, despite an increase observed with the positive control ethanol (EtOH), nor was there an increase in embryonic oxidatively damaged DNA, quantified as 8-oxo-2'-deoxyguanosine (8-oxodG) formation. MeOH teratogenicity (primarily ophthalmic anomalies, cleft palate) also was not altered by pre- and post-treatment with varying doses of the free radical spin trapping agent alpha-phenyl-N-tert-butylnitrone (PBN). In contrast, pretreatment with L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of glutathione (GSH) synthesis, depleted maternal hepatic and embryonic GSH, and enhanced some new anomalies (micrognathia, agnathia, short snout, fused digits, cleft lip, low set ears), but not the most common teratogenic effects of MeOH (ophthalmic anomalies, cleft palate) in this strain. These results suggest that ROS did not contribute to the teratogenic effects of MeOH in this in vivo mouse model, in contrast to results in embryo culture from our laboratory, and that the protective effect of GSH in this model may arise from its role as a cofactor for formaldehyde dehydrogenase in the detoxification of formaldehyde.

Ultrasound-guided suprascapular nerve block: a correlation with fluoroscopic and cadaveric findings.

PURPOSE: Previous work on the ultrasound-guided injection technique and the sonoanatomy of the suprascapular region relevant to the suprascapular nerve (SSN) block suggested that the ultrasound scan showed the presence of the suprascapular notch and transverse ligament. The intended target of the ultrasound-guided injection was the notch. The objective of this case report and the subsequent cadaver dissection findings is to reassess the interpretation of the ultrasound images when locating structures for SSN block. CLINICAL FEATURES: A 45-yr-old man with chronic shoulder pain received an ultrasound-guided SSN block using the suprascapular notch as the intended target. The position of the needle was verified by fluoroscopy, which showed the tip of the needle well outside the suprascapular notch. Similar ultrasound-guided SSN blocks were performed in two cadavers. Dissections were performed which showed that the needle tips were not at the suprascapular notch but, more accurately, were close to the SSN but at the floor of the suprascapular fossa between the suprascapular and spinoglenoid notch. CONCLUSION: Our fluoroscopic and cadaver dissection findings both suggest that the ultrasound image of the SSN block shown by the well-described technique is actually targeting the nerve on the floor of the suprascapular spine between the suprascapular and spinoglenoid notches rather than the suprascapular notch itself. The structure previously identified as the transverse ligament is actually the fascia layer of the supraspinatus muscle.

Multiscale control of flooding and riparian-forest composition in Lower Michigan, USA.

Despite general agreement that river-valley hydrology shapes riparian ecosystems, relevant processes are difficult to distinguish and often inadequately specified in riparian studies. We hypothesize that physical constraints imposed by broad-scale watershed characteristics and river valleys modify local site conditions in a predictable and probabilistic fashion. To test this hypothesis, we employ a series of structural equations that decompose occurrence of riparian ecotypes into regional temperature, catchment storm response, valley hydraulics, and local site wetness via a priori specification of factor structure and ask (1) Is there evidence for multiscale hydrologic control of riparian diversity across Lower Michigan? (2) Do representations of key constraints on flood dynamics distinguish regional patterns of riparian vegetation? (3) How important are these effects? Cross-correlation among geospatial predictors initially obscured much of the variation revealed through analysis of semipartial variance. Causal relationships implied by our model fit with observed variation in riparian conditions (chi-square P = 0.43) and accounted for between 84% and 99% of the occurrence probability of five riparian ecotypes at 94 locations. Results suggest strong variation in the effects of regional climate, and both the relative importance and spatial scale of hydrologic factors influencing riparian vegetation through explicit quantification of relative flood frequency, duration, intensity, and relative overall inundation. Although climate and hydrology are not the only determinants of riparian conditions, interactions of hydrologic sourcing and flood dynamics described by our spatial models drive a significant portion of the variation in riparian ecosystem character throughout Lower Michigan, USA.

Molecular and biochemical mechanisms in teratogenesis involving reactive oxygen species.

Developmental pathologies may result from endogenous or xenobiotic-enhanced formation of reactive oxygen species (ROS), which oxidatively damage cellular macromolecules and/or alter signal transduction. This minireview focuses upon several model drugs (phenytoin, thalidomide, methamphetamine), environmental chemicals (benzo[a]pyrene) and gamma irradiation to examine this hypothesis in vivo and in embryo culture using mouse, rat and rabbit models. Embryonic prostaglandin H synthases (PHSs) and lipoxygenases bioactivate xenobiotics to free radical intermediates that initiate ROS formation, resulting in oxidation of proteins, lipids and DNA. Oxidative DNA damage and embryopathies are reduced in PHS knockout mice, and in mice treated with PHS inhibitors, antioxidative enzymes, antioxidants and free radical trapping agents. Thalidomide causes embryonic DNA oxidation in susceptible (rabbit) but not resistant (mouse) species. Embryopathies are increased in mutant mice deficient in the antioxidative enzyme glucose-6-phosphate dehydrogenase (G6PD), or by glutathione (GSH) depletion, or inhibition of GSH peroxidase or GSH reductase. Inducible nitric oxide synthase knockout mice are partially protected. Inhibition of Ras or NF-kB pathways reduces embryopathies, implicating ROS-mediated signal transduction. Atm and p53 knockout mice deficient in DNA damage response/repair are more susceptible to xenobiotic or radiation embryopathies, suggesting a teratological role for DNA damage, consistent with enhanced susceptibility to methamphetamine in ogg1 knockout mice with deficient repair of oxidative DNA damage. Even endogenous embryonic oxidative stress carries a risk, since untreated G6PD- or ATM-deficient mice have increased embryopathies. Thus, embryonic processes regulating the balance of ROS formation, oxidative DNA damage and repair, and ROS-mediated signal transduction may be important determinants of teratological risk.

The peroxynitrite pathway in development: phenytoin and benzo[a]pyrene embryopathies in inducible nitric oxide synthase knockout mice.

Nitric oxide generated by nitric oxide synthases (NOSs) can react with reactive oxygen species (ROS), forming peroxynitrite, which may contribute to the ROS-initiated macromolecular damage implicated in the embryopathic effects of both endogenous and drug-enhanced oxidative stress. Inducible NOS (iNOS) is nonconstitutive in most tissues, and its embryonic expression and developmental importance are unknown. Herein, during organogenesis (Gestational Days 9 and 10), wild-type B6129PF2 embryos in culture were highly susceptible to the ROS-initiating teratogens phenytoin and benzo[a]pyrene, whereas iNOS knockout embryos were substantially but not completely protected (p < .05), implicating iNOS in the embryopathic mechanism. However, in contrast to prostaglandin H synthase-catalyzed teratogen bioactivation and ROS formation, which occurs within the embryo, in vivo iNOS expression was limited to placental tissue. These results suggest that the diffusion of nitric oxide from placental progenitor tissue (ectoplacental cone) to embryonic target tissues contributes to the embryopathic effects of ROS-initiating teratogens in embryo culture, which may constitute a mechanism by which embryonic determinants of ROS-mediated teratogenesis can be modulated by maternal extra-embryonic processes.

Methanol teratogenicity in mutant mice with deficient catalase activity and transgenic mice expressing human catalase.

The role of catalase in methanol (MeOH) teratogenesis is unclear. In rodents it both detoxifies reactive oxygen species (ROS) and metabolizes MeOH and its formic acid (FA) metabolite. We treated pregnant mice expressing either high (hCat) or low catalase activity (aCat), or their wild-type (WT) controls, with either MeOH (4g/kg ip) or saline. hCat mice and WTs were similarly susceptible to MeOH-initiated ophthalmic abnormalities and cleft palates. aCat and WT mice appeared resistant, precluding assessment of the developmental impact of catalase deficiency. Catalase activity was respectively increased at least 1.5-fold, and decreased by at least 35%, in hCat and aCat embryos and maternal livers. MeOH and FA pharmacokinetic profiles were similar among hCat, aCat and WT strains. Although the hCat results imply no ROS involvement, embryo culture studies suggest this may be confounded by maternal factors and/or a requirement for higher catalase activity in the hCat mice.

Fluorothalidomide: a characterization of maternal and developmental toxicity in rabbits and mice.

The expanding therapeutic uses of thalidomide (TD) are limited by its teratogenic side effects. Although the therapeutic and teratogenic effects may be stereoselectively separable, rapid in vivo racemization of the TD isomers confounds the corroboration of this distinction. Herein we evaluated the potential of fluorothalidomide (FTD), the closest structural analog of TD with stable, nonracemizing isomers, as a model compound for studying stereoselectivity in TD teratogenesis. In contrast to TD, FTD was a potent maternal and fetal toxicant in both rabbits and mice in vivo. Furthermore, FTD rapidly degraded in vivo, presumably via hydrolysis, which in vitro was up to 22-fold faster for FTD than TD. Most critically, in an established rabbit embryo culture model for TD teratogenesis, FTD did not initiate the limb bud embryopathies observed with TD. The chemical instability and strikingly different maternal and developmental toxicological profiles of FTD and TD make FTD an unsuitable compound for studying stereoselective mechanisms of TD teratogenesis.

Species- and strain-dependent teratogenicity of methanol in rabbits and mice.

Estimates of human risk for developmental toxicity of methanol (MeOH) are based on studies in rodents, which unlike humans use catalase to metabolize MeOH. Rabbits, like humans, may largely use alcohol dehydrogenase (ADH), and more accurately than rodents reflect primate MeOH and formic acid (FA) pharmacokinetic profiles. Here we show that New Zealand white rabbits and one strain of mouse (C3H) are resistant to MeOH teratogenicity, whereas C57BL/6J mice are susceptible. Neither rabbits nor mice were susceptible to the acute MeOH toxicity observed in humans. The strain-dependent teratological susceptibility in mice could not be explained by differences in MeOH or FA disposition, nor could the resistance of rabbits, which exhibited more prolonged FA accumulation, suggesting that different mechanisms underlie MeOH teratogenesis and the FA-mediated acute toxicity in humans. It is not clear if the human risk for MeOH developmental toxicity can be accurately estimated using sensitive rodent strains.

Oxidative stress in developmental origins of disease: teratogenesis, neurodevelopmental deficits, and cancer.

In the developing embryo and fetus, endogenous or xenobiotic-enhanced formation of reactive oxygen species (ROS) like hydroxyl radicals may adversely alter development by oxidatively damaging cellular lipids, proteins and DNA, and/or by altering signal transduction. The postnatal consequences may include an array of birth defects (teratogenesis), postnatal functional deficits, and diseases. In animal models, the adverse developmental consequences of in utero exposure to agents like thalidomide, methamphetamine, phenytoin, benzo[a]pyrene, and ionizing radiation can be modulated by altering pathways that control the embryonic ROS balance, including enzymes that bioactivate endogenous substrates and xenobiotics to free radical intermediates, antioxidative enzymes that detoxify ROS, and enzymes that repair oxidative DNA damage. ROS-mediated signaling via Ras, nuclear factor kappa B and related transducers also may contribute to altered development. Embryopathies can be reduced by free radical spin trapping agents and antioxidants, and enhanced by glutathione depletion. Further modulatory approaches to evaluate such mechanisms in vivo and/or in embryo culture have included the use of knockout mice, transgenic knock-ins and mutant deficient mice with altered enzyme activities, as well as antisense oligonucleotides, protein therapy with antioxidative enzymes, dietary depletion of essential cofactors and chemical enzyme inhibitors. In a few cases, measures anticipated to be protective have conversely enhanced the risk of adverse developmental outcomes, indicating the complexity of development and need for caution in testing therapeutic strategies in humans. A better understanding of the developmental effects of ROS may provide insights for risk assessment and the reduction of adverse postnatal consequences.

A GIS model of subsurface water potential for aquatic resource inventory, assessment, and environmental management.

Biological, chemical, and physical attributes of aquatic ecosystems are often strongly influenced by groundwater sources. Nonetheless, widespread access to predictions of subsurface contributions to rivers, lakes, and wetlands at a scale useful to environmental managers is generally lacking. In this paper, we describe a "neighborhood analysis" approach for estimating topographic constraints on spatial patterns of recharge and discharge and discuss how this index has proven useful in research, management, and conservation contexts. The Michigan Rivers Inventory subsurface flux model (MRI-DARCY) used digital elevation and hydraulic conductivity inferred from mapped surficial geology to estimate spatial pattems of hydraulic potential. Model predictions were calculated in units of specific discharge (meters per day) for a 30-m-cell raster map and interpreted as an index of potential subsurface water flux (shallow groundwater and event through-flow). The model was evaluated by comparison with measurements of groundwater-related attributes at watershed, stream segment, and local spatial scales throughout Lower Michigan (USA). Map-based predictions using MRI-DARCY accounted for 85% of the observed variation in base flow from 128 USGS gauges, 69% of the observed variation in discharge accrual from 48 river segments, and 29% of the residual variation in local groundwater flux from 33 locations as measured by hyporheic temperature profiles after factoring out the effects of climate. Although it does not incorporate any information about the actual water table surface, by quantifying spatial variation of key constraints on groundwater-related attributes, the model provides strata for more intensive study, as well as a useful spatial tool for regional and local conservation planning, fisheries management, wetland characterization, and stream assessment.