Impact of hydrologic boundaries on microbial planktonic and biofilm communities in shallow terrestrial subsurface environments.

Subsurface environments contain a large proportion of planetary microbial biomass and harbor diverse communities responsible for mediating biogeochemical cycles important to groundwater used by human society for consumption, irrigation, agriculture and industry. Within the saturated zone, capillary fringe and vadose zones, microorganisms can reside in two distinct phases (planktonic or biofilm), and significant differences in community composition, structure and activity between free-living and attached communities are commonly accepted. However, largely due to sampling constraints and the challenges of working with solid substrata, the contribution of each phase to subsurface processes is largely unresolved. Here, we synthesize current information on the diversity and activity of shallow freshwater subsurface habitats, discuss the challenges associated with sampling planktonic and biofilm communities across spatial, temporal and geological gradients, and discuss how biofilms may be constrained within shallow terrestrial subsurface aquifers. We suggest that merging traditional activity measurements and sequencing/-omics technologies with hydrological parameters important to sediment biofilm assembly and stability will help delineate key system parameters. Ultimately, integration will enhance our understanding of shallow subsurface ecophysiology in terms of bulk-flow through porous media and distinguish the respective activities of sessile microbial communities from more transient planktonic communities to ecosystem service and maintenance.

Novel Semiquantitative Bone Marrow Oedema Score and Fracture Score for the Magnetic Resonance Imaging Assessment of the Active Charcot Foot in Diabetes.

There are no accepted methods to grade bone marrow oedema (BMO) and fracture on magnetic resonance imaging (MRI) scans in Charcot osteoarthropathy. The aim was to devise semiquantitative BMO and fracture scores on foot and ankle MRI scans in diabetic patients with active osteoarthropathy and to assess the agreement in using these scores. Three radiologists assessed 45 scans (Siemens Avanto 1.5T, dedicated foot and ankle coil) and scored independently twenty-two bones (proximal phalanges, medial and lateral sesamoids, metatarsals, tarsals, distal tibial plafond, and medial and lateral malleoli) for BMO (0-no oedema, 1-oedema < 50% of bone volume, and 2-oedema > 50% of bone volume) and fracture (0-no fracture, 1-fracture, and 2-collapse/fragmentation). Interobserver agreement and intraobserver agreement were measured using multilevel modelling and intraclass correlation (ICC). The interobserver agreement for the total BMO and fracture scores was very good (ICC = 0.83, 95% confidence intervals (CI) 0.76, 0.91) and good (ICC = 0.62; 95% CI 0.48, 0.76), respectively. The intraobserver agreement for the total BMO and fracture scores was good (ICC = 0.78, 95% CI 0.6, 0.95) and fair to moderate (ICC = 0.44; 95% CI 0.14, 0.74), respectively. The proposed BMO and fracture scores are reliable and can be used to grade the extent of bone damage in the active Charcot foot.

Mechanisms of injury and CT findings in bowel and mesenteric trauma.

Bowel and mesenteric injuries are relatively uncommon but associated with significant morbidity and mortality. Early recognition is crucial, and multidetector computed tomography (MDCT) now has a central role in the evaluation of patients with a history of trauma. In this review, we describe the MDCT appearances of bowel and mesenteric injuries with reference to findings at surgery. Emphasis is placed on the importance of an understanding of mechanism of injury when interpreting CT findings following abdominal trauma.

The value of ultrasound in assessing displacement of a medial clavicular physeal separation in an adolescent.

We describe a case of medial clavicular physeal separation with posterior displacement of the metaphysis in a 13-year-old girl, focusing on the role played by ultrasound in the diagnosis, planning of treatment and post-reduction follow-up. On clinical examination and conventional radiography, the injury is essentially indistinguishable from a sternoclavicular dislocation or a fracture of the medial aspect of the clavicle; however, the pathogenesis is different, consisting in medial physeal separation and 'degloving' of the inner cancellous bone of the metaphysis from the surrounding periosteal collar with posterior metaphyseal dislocation. In our case, attempted closed reduction failed, and the injury required open reduction with relocation of the clavicle into the periosteal sleeve followed by suturing of the periosteal tear.

Generalized schemes for high-throughput manipulation of the Desulfovibrio vulgaris genome.

The ability to conduct advanced functional genomic studies of the thousands of sequenced bacteria has been hampered by the lack of available tools for making high-throughput chromosomal manipulations in a systematic manner that can be applied across diverse species. In this work, we highlight the use of synthetic biological tools to assemble custom suicide vectors with reusable and interchangeable DNA "parts" to facilitate chromosomal modification at designated loci. These constructs enable an array of downstream applications, including gene replacement and the creation of gene fusions with affinity purification or localization tags. We employed this approach to engineer chromosomal modifications in a bacterium that has previously proven difficult to manipulate genetically, Desulfovibrio vulgaris Hildenborough, to generate a library of over 700 strains. Furthermore, we demonstrate how these modifications can be used for examining metabolic pathways, protein-protein interactions, and protein localization. The ubiquity of suicide constructs in gene replacement throughout biology suggests that this approach can be applied to engineer a broad range of species for a diverse array of systems biological applications and is amenable to high-throughput implementation.

Classification of the morphology of the acromioclavicular joint using cadaveric and radiological analysis.

The aim of this study was to establish a classification system for the acromioclavicular joint using cadaveric dissection and radiological analyses of both reformatted computed tomographic scans and conventional radiographs centred on the joint. This classification should be useful for planning arthroscopic procedures or introducing a needle and in prospective studies of biomechanical stresses across the joint which may be associated with the development of joint pathology. We have demonstrated three main three-dimensional morphological groups namely flat, oblique and curved, on both cadaveric examination and radiological assessment. These groups were recognised in both the coronal and axial planes and were independent of age.

Imaging of patellofemoral disorders.

Anterior knee pain is a common symptom, which may have a large variety of causes including patellofemoral pathologies. Patellofemoral maltracking refers to dynamic abnormality of patellofemoral alignment and has been measured using plain film, computed tomography (CT) and magnetic resonance imaging (MRI) using static and kinematic techniques. Patellar dislocation is usually transient, but specific conventional radiographic and MRI features may provide evidence of prior acute or chronic dislocation. In addition, chondromalacia patellae, osteochondritis dissecans, patellofemoral osteoarthritis, excessive lateral pressure syndrome, and bipartite patella have all been implicated in causing patellofemoral pain. The imaging and clinical features of these processes are reviewed, highlighting the specific diagnostic features of each condition.

Characterization of microbial activities and U reduction in a shallow aquifer contaminated by uranium mill tailings.

A characterization of the Shiprock, NM, uranium mill tailing site focused on the geochemical and microbiological factors governing in-situ uranium-redox reactions. Groundwater and aqueous extracts of sediment samples contained a wide concentration range of sulfate, nitrate, and U(VI) with median values of 21.2 mM, 16.1 micro M, and 2.7 micro M, respectively. Iron(III) was not detected in groundwater, but a median value of 0.3 mM in sediment extracts was measured. Bacterial diversity down gradient from the disposal pile reflected the predominant geochemistry with relatively high numbers of sulfate- and nitrate-reducing microorganisms, and smaller numbers of acetogenic, methanogenic, nitrate-dependent Fe(II)-oxidizing, Fe(III)-reducing, and sulfide-oxidizing bacteria. In aquifer slurry incubations, nitrate reduction was always preferred and had a negative impact on sulfate-, Fe(III)-, and U-reduction rates. We also found that sulfate-reduction rates decreased sharply in the presence of clay, while Fe(III)-reduction increased with no clear impact on U reduction. In the absence of clay, iron and sulfate reduction correlated with concentrations of Fe(III) and sulfate, respectively. Rates of U(VI) loss did not correlate with the concentration of any electron acceptor. With the exception of Fe(III), electron donor amendment was largely unsuccessful in stimulating electron acceptor loss over a 2-week incubation period, suggesting that endogenous forms of organic matter were sufficient to support microbial activity. Our findings suggest that efforts to accelerate biological U reduction should initially focus on stimulating nitrate removal.

Purification of the NADP+:F420 oxidoreductase of Methanosphaera stadtmanae.

Methanosphaera stadtmanae (DSM 3091) is a methanogen that requires H2 and CH3OH for methanogenesis. The organism does not possess an F420-dependent hydrogenase and only low levels of F420. It does however possess NADP+:F420 oxidoreductase activity. The NADP+:F420 oxidoreductase, the enzyme which catalyses the electron transfer between NADP+ and F420 in this organism, was purified and characterized. NAD+, NADH, FMN, and FAD could not be used as electron acceptors. Optimal pH for F420 reduction was 6.0, and 8.5 for NADP+ reduction. During the purification process, it was noted that precipitation with (NH4)2SO4 increased total activity 16-fold but reduced the stability of the enzyme. However, recombination of cell-free extracts with resuspended 65-90% (NH4)2SO4 pellet returned activity to near cell-free extract levels. Neither high salt or protease inhibitors were effective in stabilizing the activity of the partially purified enzyme. The purified enzyme from M. stadtmanae possessed a molecular weight of 148 kDa as determined by gel filtration chromatography and native-PAGE, consisting of alpha, beta, and gamma subunits of 60, 50, and 45 kDa, respectively, using SDS-PAGE. The Km values were 370 microM for NADP+, 142 microM for NADPH, 62.5 microM for F420, and 7.7 microM for F420H2. These values were different from the Km values observed in the cell-free extract.

Estimation of methanogen biomass by quantitation of coenzyme M.

Determination of the role of methanogenic bacteria in an anaerobic ecosystem often requires quantitation of the organisms. Because of the extreme oxygen sensitivity of these organisms and the inherent limitations of cultural techniques, an accurate biomass value is very difficult to obtain. We standardized a simple method for estimating methanogen biomass in a variety of environmental matrices. In this procedure we used the thiol biomarker coenzyme M (CoM) (2-mercaptoethanesulfonic acid), which is known to be present in all methanogenic bacteria. A high-performance liquid chromatography-based method for detecting thiols in pore water (A. Vairavamurthy and M. Mopper, Anal. Chim. Acta 78:363-370, 1990) was modified in order to quantify CoM in pure cultures, sediments, and sewage water samples. The identity of the CoM derivative was verified by using liquid chromatography-mass spectroscopy. The assay was linear for CoM amounts ranging from 2 to 2,000 pmol, and the detection limit was 2 pmol of CoM/ml of sample. CoM was not adsorbed to sediments. The methanogens tested contained an average of 19.5 nmol of CoM/mg of protein and 0.39 +/- 0.07 fmol of CoM/cell. Environmental samples contained an average of 0.41 +/- 0.17 fmol/cell based on most-probable-number estimates. CoM was extracted by using 1% tri-(N)-butylphosphine in isopropanol. More than 90% of the CoM was recovered from pure cultures and environmental samples. We observed no interference from sediments in the CoM recovery process, and the method could be completed aerobically within 3 h. Freezing sediment samples resulted in 46 to 83% decreases in the amounts of detectable CoM, whereas freezing had no effect on the amounts of CoM determined in pure cultures. The method described here provides a quick and relatively simple way to estimate methanogenic biomass.

Hypercapnia lowers the shivering threshold and increases core cooling rate in humans.

BACKGROUND: Hypercapnia, which may be encountered during diving operations or by patients under general anesthesia with spontaneous respiration, alters the basic thermoregulatory responses of animals and humans. In cold-exposed animals, 3-10% inspired CO2 impairs thermal homeostasis by attenuating shivering and promoting heat loss through peripheral vasodilation. Experimental results with humans are equivocal. PURPOSE: To determine the effects of hypercapnia on warm and cold thermoregulatory responses and core cooling rates during mild cold stress, we examined the core temperature (TCO) thresholds for sweating, vasoconstriction and shivering, as well as core cooling rates of eight subjects immersed in 28 degrees C water under hypercapnic conditions. METHODS: On two separate days, subjects exercised on an underwater cycle ergometer to elevate TCO above the sweating threshold. They then rested and cooled until they shivered vigorously. Subjects inspired humidified room air during the control trial and 4% CO2/20.9% O2/balance N2 during the hypercapnia trial. RESULTS: Hypercapnia lowered the threshold for shivering by 0.13 degrees C and increased the core cooling rate by 0.35 degrees C.h-1 (25%). Minute ventilation was approximately 12.0 L.min-1 throughout cooling during control and increased from 25.3 to 28.7 L.min-1 during hypercapnia. CONCLUSIONS: Therefore, hypercapnia enhances the core cooling rate during mild cold stress. This may be attributed in part to a delay in shivering onset, as well as increased respiratory heat loss during hypercapnic hyperventilation.

Alcohol lowers the vasoconstriction threshold in humans without affecting core cooling rate during mild cold exposure.

Elevated blood alcohol levels are often seen in hypothermia and hyperthermia related deaths, leading to the belief that alcohol renders humans poikilothermic. We examined the core temperature (Tco) thresholds for sweating, vasoconstriction and shivering as well as core cooling rates of seven subjects immersed in 28 degrees C water. On two separate days, subjects exercised on an underwater cycle ergometer to elevate Tco above the sweating threshold. They then rested and cooled until they shivered vigorously. Subjects drank orange juice (7 ml.kg-1) prior to immersion during the control trial and 1 ml.kg-1 absolute ethanol, added to orange juice in a 1:6 ratio, during the alcohol trial. Mean blood alcohol concentration (breath analysis) was 0.097 +/- 0.010 g% at the start of cooling and 0.077 +/- 0.008 g% at the end of the cooling period. Alcohol lowered the vasoconstriction threshold by 0.32 +/- 0.2 degrees C and elevated finger tip blood flow, but had no effect on thresholds for sweating and shivering or core cooling rate. Considering these minor effects it is unlikely that moderate alcohol consumption predisposes individuals to hypothermia or hyperthermia via impaired thermoregulation, but rather likely due to behavioral factors.

MRI and CT of sellar and parasellar disorders.

Discussion of the anatomy of the parasellar region focuses on the optic chiasm and its relationship to surrounding structures. CT and MRI are valuable tools in the imaging of these regions. This article provides a brief review of the technique of CT and MRI with visualized anatomic structures of these areas. A closer look at how CT and MRI are used to delineate the different pathologic processes of the sellar and parasellar regions then follows.

Optic nerve tumors: role of magnetic resonance imaging and computed tomography.

Localization and pathologic diagnosis of optic nerve tumors have been greatly improved by high resolution computed tomography and magnetic resonance imaging. Radiologic differentiation of the most common tumors of the optic nerve (meningioma and glioma) can be made by using these new imaging modalities. Early diagnosis of optic nerve lesions has resulted in better prognosis and surgical outcome.