Immobilization of Cr(VI) and its reduction to Cr(III) phosphate by granular biofilms comprising a mixture of microbes.

We assessed the potential of mixed microbial consortia, in the form of granular biofilms, to reduce chromate and remove it from synthetic minimal medium. In batch experiments, acetate-fed granular biofilms incubated aerobically reduced 0.2 mM Cr(VI) from a minimal medium at 0.15 mM day(-1) g(-1), with reduction of 0.17 mM day(-1) g(-1) under anaerobic conditions. There was negligible removal of Cr(VI) (i) without granular biofilms, (ii) with lyophilized granular biofilms, and (iii) with granules in the absence of an electron donor. Analyses by X-ray absorption near edge spectroscopy (XANES) of the granular biofilms revealed the conversion of soluble Cr(VI) to Cr(III). Extended X-ray absorption fine-structure (EXAFS) analysis of the Cr-laden granular biofilms demonstrated similarity to Cr(III) phosphate, indicating that Cr(III) was immobilized with phosphate on the biomass subsequent to microbial reduction. The sustained reduction of Cr(VI) by granular biofilms was confirmed in fed-batch experiments. Our study demonstrates the promise of granular-biofilm-based systems in treating Cr(VI)-containing effluents and wastewater.

Quantification of tantalum sedimentation rates in liquid embolic agents.

Background Endovascular treatment (EVT) of brain arteriovenous malformations has evolved from cyanoacrylate derivatives such as N-butyl cyanoacrylate, an adhesive glue, to ethylene vinyl copolymer-based liquid embolics such as Onyx® and SQUID® dissolved in dimethyl sulfoxide. Although these agents offer several advantages, their rapidly decreasing radiopacity, as a result of the sedimentation of tantalum powder, compromises visual control during EVT. This study aims to quantify and compare tantalum sedimentation rates of several liquid embolic agents, and determine their effects on radiopacity. Methods The rate of sedimentation of liquid embolics Onyx 18®, SQUID 12®, and SQUID 18® was measured after preparation by single x-ray exposures for a period of 30 minutes. The signal-to-noise ratios (SNRs) of the suspension of each liquid embolic was calculated at various time points as tantalum settled out of the suspension. Precipitating Hydrophobic Injectable Liquid (PHIL®) was imaged as a control. Results Onyx 18® demonstrated the fastest sedimentation rate of the liquid embolics analyzed and demonstrated a threefold faster drop in SNR compared to SQUID 18® over 30 minutes. Onyx 18® demonstrated a one and a half times faster drop in SNR compared to SQUID 12®. Although PHIL 25® maintained constant SNR over the same time, it was lower at baseline immediately after preparation compared to tantalum-based liquids. Conclusion Caution during long injections using tantalum-based agents is advised. Onyx 18® has a significantly faster drop in radiopacity compared to SQUID 12® and SQUID 18®. Covalently bonded iodine-based embolics like PHIL® demonstrate constant radiopacity over time.

Microbial mobilization of plutonium and other actinides from contaminated soil.

We examined the dissolution of Pu, U, and Am in contaminated soil from the Nevada Test Site (NTS) due to indigenous microbial activity. Scanning transmission x-ray microscopy (STXM) analysis of the soil showed that Pu was present in its polymeric form and associated with Fe- and Mn- oxides and aluminosilicates. Uranium analysis by x-ray diffraction (µ-XRD) revealed discrete U-containing mineral phases, viz., schoepite, sharpite, and liebigite; synchrotron x-ray fluorescence (µ-XRF) mapping showed its association with Fe- and Ca-phases; and µ-x-ray absorption near edge structure (µ-XANES) confirmed U(IV) and U(VI) oxidation states. Addition of citric acid or glucose to the soil and incubated under aerobic or anaerobic conditions enhanced indigenous microbial activity and the dissolution of Pu. Detectable amount of Am and no U was observed in solution. In the citric acid-amended sample, Pu concentration increased with time and decreased to below detection levels when the citric acid was completely consumed. In contrast, with glucose amendment, Pu remained in solution. Pu speciation studies suggest that it exists in mixed oxidation states (III/IV) in a polymeric form as colloids. Although Pu(IV) is the most prevalent and generally considered to be more stable chemical form in the environment, our findings suggest that under the appropriate conditions, microbial activity could affect its solubility and long-term stability in contaminated environments.

Bidirectional gene transfer between Aspergillus fumigatus and Aspergillus nidulans.

A genomic DNA library was constructed from a pathogenic strain of Aspergillus fumigatus using the cosmid vector pCosAX. Cosmid clones with homologies to the rodA, brlA, fluG, flbA or trpC genes from A. nidulans were isolated from the library. Each A. fumigatus clone was used to complement a strain of A. nidulans with a mutation in the homologous gene. A spontaneous white spored strain of A. fumigatus was isolated. The mutation was complemented by transforming the strain with a plasmid containing the wA gene from A. nidulans. The results from these experiments indicate a significant degree of structural and functional homology between genes from the organisms. These findings indicate the potential to exploit the methods and information available from A. nidulans to address questions related to human disease caused by A. fumigatus and the ability to use A. nidulans as a surrogate genetic host for characterizing A. fumigatus gene function.

The chsA gene from Aspergillus nidulans is necessary for maximal conidiation.

A fragment from the open reading frame of the cloned chsA gene from Aspergillus nidulans was deleted and replaced with the argB gene. The resulting construct was used to replace the wild-type chsA gene in an argB deletion strain. The growth and morphology of the vegetative hyphae from the resulting chsA disruptant strain were indistinguishable from those of a wild-type strain but the chitin content of the hyphae from the disruptant was reduced to approximately 90% of that of wild-type. The disruptant showed reduced ability to produce the asexual spores (conidia) that are formed by differentiated aerial hyphae called conidiophores. The ability to form undifferentiated aerial hyphae was not impaired in the disruptant. The conidiophores and conidia produced by the disruptant were indistinguishable from those of wild-type. Conidium formation by the disruptant grown on a variety of media was reduced to about 30% of the wild-type. A chsE null strain did not show a defect in conidiation but a strain in which both chsA and chsE were inactivated produced about 3% of the conidia of wild-type. That finding supports the hypothesis that chsA and chsE encode a partially redundant function necessary for conidiophore development.

Morphine induced increases in the incorporation of 3H-thymidine into brain striatal DNA.

3H-thymidine uptake into DNA fractions of rat brain regions was measured following in vivo administration of (methyl-3H)-thymidine and morphine. Acute morphine administration (10 mg/kg; 30 min prior to 3H-thymidine) increased incorporation of 3H-thymidine into DNA of rat striatum. This effect was antagonized by naloxone (1 mg/kg). Further, the observed change in incorporation of 3H-thymidine into DNA in striatum could not be accounted for by differences in the local availability of the label in morphinized rats. An autoradiographic study revealed that the 3H-thymidine was localized in nuclei in cells of the sub-ependymal layer lining the lateral ventricles, an area of glial cell proliferation in adult rats. No change in 3H-thymidine incorporation into DNA was observed in any area of the brain in morphine-addicted rats or in rats undergoing naloxone-precipitated withdrawal. The results indicate that opiates may induce permanent anatomical changes in the brain, including alterations of neuroglial interactions.

Uganda--rehabilitation, or redefinition of health services?

Uganda had one of the best health care delivery systems in Africa. The decade of misrule by Amin saw a collapse of the country and an exodus of doctors and other professions. The 1979 liberation war and subsequent political instability and insecurity further aggravated the poor health services then available. When political stability was temporarily restored in December 1980 the cash crop export sector took priority over social services and the health budget declined to only 3.5% compared to a former level of 7.5% of government budget. Emergencies in West Nile, Karamoja and the Luwero triangle continued to plague rehabilitation efforts upto 1985. Alternate strategies for improving health are proposed including female education, increased budget allocations, food and nutrition policy and health information. Uganda's prospect for rebuilding the health services has begun with immunization, control of diarrhoeal diseases, nutrition surveillance in Karamoja and an essential drugs programme, but the success of these is dependent upon political stability and improvement in overall security.

Health implications of war in Uganda and Sudan.

Civil war disrupted agriculture and trade in Uganda and Sudan. This reduced tax revenues and drained scarce resources away from health budgets to finance increased military expenditures. Hundreds and thousands of people were driven from their homes either as internally displaced people or as refugees. Normal health service delivery systems were broken down forcing doctors, nurses and other health professionals into towns, cities or neighbouring countries in search of peace and employment. Scores of hospitals, health centres and dispensaries were abandoned, destroyed or looted, rendering even the limited physical facilities useless. Preventive public health services such as immunization and provision of potable drinking water were discontinued leaving huge populations susceptible to controllable infectious diseases and epidemics.

Effect of stepping rate on energy costs during StairMaster exercise.

The responses to a self-selected stepping pattern (random) on a StairMaster 4000PT were compared with those obtained in response to the rates established by the manufacturer (cadence) in men (N = 14) and women (N = 14). During the random test the subjects stepped at their own natural, self-selected rate and distance. In cadence trial the subjects were required to step in time with a metronome at a predetermined rates of 60, 77, 95, and 112 steps.min-1. Each trial consisted of four, 5-min continuous workloads during which HRs were recorded and expired air was analyzed using an automated open-circuit gas system each minute. All size dependent variables (i.e., VE and lO2.min-1) as well as relative VO2 (mlO2.kg-1.min-1) were significantly (P < 0.01) higher for the men across all stages and between methods. Although the random test produced slightly higher oxygen consumption values than the cadence trial, these differences were not significant (P > 0.05). The actual METs were significantly (P < 0.01) higher at all stages except at the lowest stepping rate for both methods compared with those estimated by the manufacturer. Equations were established to estimate actual MET costs: Men's METs = 2.675 + 0.935 (rate); women's METs = 2.934 + 0.817 (rate). Cross-validations of 0.975 and 0.957 were obtained on an additional group of men (N = 8) and women (N = 11), respectively.

Emergence characteristics of sevoflurane compared to halothane in pediatric patients undergoing bilateral pressure equalization tube insertion.

STUDY OBJECTIVE: To compare the emergence characteristics of sevoflurane with halothane in pediatric patients having bilateral myringotomy and pressure equalization tube insertion using a highly standardized and common anesthetic technique. DESIGN: Prospective, randomized, double-blind study. SETTING: University hospital. PATIENTS: 43 ASA physical status I and II healthy pediatric outpatients scheduled for initial ear tube placement by one surgeon. INTERVENTIONS: Patients were randomized to receive either halothane or sevoflurane as their sole anesthetic drug for ear tube insertion. All patients as well as the research nurse grading emergence agitation were blinded to group assignment. No premedication was administered, and all patients received a standard dose of rectal acetaminophen for postoperative pain control. MEASUREMENT AND MAIN RESULTS: The primary outcome was the percentage of patients with emergence agitation, defined as thrashing behavior requiring physical restraint for greater than three minutes. Time to discharge from the postanesthesia care unit (PACU) and from the hospital were also compared. Sevoflurane patients had a greater incidence of emergence delirium: 57% versus 27% [95% CI (1.7%-58.1%) p = 0.047]. Time to discharge from the PACU and the hospital were longer for patients who received sevoflurane: 62 versus 50 min [95% CI (1.4-23.6) p = 0.02] and 102 versus 79 mins [95% CI (9.6-37.1) p = 0.003]. CONCLUSION: In our institution there is a trend toward greater emergence agitation in ear tube placement patients given sevoflurane versus halothane anesthesia. Further, (with our discharge criteria) patients are actually discharged from the PACU and the hospital faster when given halothane as a sole anesthetic when compared to sevoflurane.

Parental presence plus oral midazolam decreases frequency of 5% halothane inductions in children.

STUDY OBJECTIVE: To determine the frequency of 5% halothane induction and behavioral distress during inhalation induction with both oral midazolam and parental presence compared with parental presence alone. DESIGN: Randomized, controlled, double blind study. SETTING: Same day anesthesia at a university department of anesthesiology. PATIENTS: 72 ASA status I and II children, 3 to 10 years of age, scheduled for first time anesthesia. INTERVENTIONS: Children were assigned to one of two groups to receive midazolam 0.5 mg/kg orally or placebo. A parent was present during induction. Children were videotaped at baseline, after treatment, and during induction. Tapes were scored for behavioral distress using the revised Observational Scale of Behavioral Distress. MEASUREMENTS AND MAIN RESULTS: Children who received midazolam in the setting of parental presence had significantly fewer 5% halothane inductions than those who received placebo (p < 0.02). They also had less behavioral distress (p < 0.01). CONCLUSIONS: The combination of parental presence plus oral midazolam reduces the likelihood of needing a 5% rapid halothane induction when compared with parental presence without premedication.

Bioreduction and precipitation of uranium in ionic liquid aqueous solution by Clostridium sp.

The ionic liquids, 1-butyl-3-methylimidazolium hexafluorophosphate [BMIM][PF6], N-ethylpyridiniumtrifluoroacetate [EtPy][CF3COO] and N-ethylpyridiniumtetrafluoroborate [EtPy][BF4], affected the reduction and precipitation of uranium by Clostridium sp. to a varying degree. Characterization of uranium association with the ionic liquids showed that uranium formed a monodentate complex with the anion BF4(-) and PF6(-) of [EtPy][BF4] and [BMIM][PF6], respectively; and a bidentate complex with carboxylate of [EtPy][CF3COO]. Bioreduction of U(VI) was influenced by the type of complex formed: monodentate complexes were readily reduced whereas the bidentate complex of U(VI) with [CF3COO] was recalcitrant. [EtPy][BF4] affected the rate and extent of precipitation of the reduced uranium; at higher concentration the reduced U(IV) remained in the solution phase. The results suggest that by tuning the properties of ionic liquids they may be valuable candidates for uranium biotreatment.

Bioreduction of uranium(VI) complexed with citric acid by Clostridia affects its structure and solubility.

Uranium contamination of the environment from mining and milling operations, nuclear-waste disposal, and ammunition use is a widespread global problem. Natural attenuation processes such as bacterial reductive precipitation and immobilization of soluble uranium is gaining much attention. However, the presence of naturally occurring organic ligands can affect the precipitation of uranium. Here, we report that the anaerobic spore-forming bacteria Clostridia, ubiquitous in soils, sediments, and wastes, capable of reduction of Fe(III) to Fe(II), Mn(IV) to Mn(II), U(VI) to U(IV), Pu(IV) to Pu(III), and Tc(VI) to Tc(IV); reduced U(VI) associated with citric acid in a dinuclear 2:2 U(VI): citric acid complex to a biligand mononuclear 1:2 U(IV):citric acid complex,which remained in solution, in contrast to reduction and precipitation of uranium. Our findings show that U(VI) complexed with citric acid is readily accessible as an electron acceptor despite the inability of the bacterium to metabolize the complexed organic ligand. Furthermore, it suggests that the presence of organic ligands at uranium-contaminated sites can affect the mobility of the actinide under both oxic and anoxic conditions by forming such soluble complexes.

Colloidal cutin-like substances cross-linked to siderophore decomposition products mobilizing plutonium from contaminated soils.

Relatively recently, inorganic colloids have been invoked to reconcile the apparent contradictions between expectations based on classical dissolved-phase Pu transport and field observations of "enhanced" Pu mobility (Kersting et al. Nature 1999, 397, 56-59). A new paradigm for Pu transport is mobilization and transport via biologically produced ligands. This study for the first time reports a new finding of Pu being transported, at sub-pM concentrations, by a cutin-like natural substance containing siderophore-like moieties and virtually all mobile Pu. Most likely, Pu is complexed by chelating groups derived from siderophores that are covalently bound to a backbone of cutin-derived soil degradation products, thus revealing the history of initial exposure to Pu. Features such as amphiphilicity and small size make this macromolecule an ideal collector for actinides and other metals and a vector for their dispersal. Cross-linking to the hydrophobic domains (e.g., by polysaccharides) gives this macromolecule high mobility and a means of enhancing Pu transport. This finding provides a new mechanism for Pu transport through environmental systems that would not have been predicted by Pu transport models.

Coordination modes in the formation of the ternary Am(III), Cm(III), and Eu(III) complexes with EDTA and NTA: TRLFS, 13C NMR, EXAFS, and thermodynamics of the complexation.

The formation and the structure of the ternary complexes of trivalent Am, Cm, and Eu with mixtures of EDTA+NTA (ethylenediamine tetraacetate and nitrilotriacetate) have been studied by time-resolved laser fluorescence spectroscopy, 13C NMR, extended X-ray absorption fine structure, and two-phase metal ion equilibrium distribution at 6.60 m (NaClO4) and a hydrogen ion concentration value (pcH) between 3.60 and 11.50. In the ternary complexes, EDTA binds via four carboxylates and two nitrogens, while the binding of the NTA varies with the hydrogen ion concentration, pcH, and the concentration ratios of the metal ion and the ligand. When the concentration ratios of the metal to ligand is low (1:1:1-1:1:2), two ternary complexes, M(EDTA)(NTAH)(3-) and M(EDTA)(NTA)(4-), are formed at pcH ca. 9.00 in which NTA binds via three carboxylates, via two carboxylates and one nitrogen, or via two carboxylates and a H2O. At higher ratios (1:1:20 and 1:10:10) and pcH's of ca. 9.00 and 11.50, one ternary complex, M(EDTA)(NTA)(4-), is formed in which NTA binds via three carboxylates and not via nitrogen. The two-phase equilibrium distribution studies at tracer concentrations of Am, Cm, and Eu have also confirmed the formation of the ternary complex M(EDTA)(NTA)(4-) at temperatures between 0 and 60 degrees C. The stability constants (log beta111) for these metal ions increase with increasing temperature. The endothermic enthalpy and positive entropy indicated a significant effect of cation dehydration in the formation of the ternary complexes at high ionic strength.

Analysis of novel soluble chromate and uranyl reductases and generation of an improved enzyme by directed evolution.

Most polluted sites contain mixed waste. This is especially true of the U.S. Department of Energy (DOE) waste sites which hold a complex mixture of heavy metals, radionuclides, and organic solvents. In such environments enzymes that can remediate multiple pollutants are advantageous. We report here evolution of an enzyme, ChrR6 (formerly referred to as Y6), which shows a markedly enhanced capacity for remediating two of the most serious and prevalent DOE contaminants, chromate and uranyl. ChrR6 is a soluble enzyme and reduces chromate and uranyl intracellularly. Thus, the reduced product is at least partially sequestered and nucleated, minimizing the chances of reoxidation. Only one amino acid change, (Tyr)128(Asn), was responsible for the observed improvement. We show here that ChrR6 makes Pseudomonas putida and Escherichia coli more efficient agents for bioremediation if the cellular permeability barrier to the metals is decreased.

Decontamination of uranium-contaminated steel surfaces by hydroxycarboxylic acid with uranium recovery.

We developed a simple, safe method to remove uranium from contaminated metallic surfaces so that the materials can be recycled or disposed of as low-level radioactive or nonradioactive waste. Surface analysis of rusted uranium-contaminated plain carbon-steel coupons by X-ray photoelectron spectroscopy and Rutherford backscattering spectroscopy showed that uranium was predominantly associated with ferrihydrite, lepidocrocite, and magnetite, or occluded in the matrix of the corrosion product as uranyl hydroxide and schoepite (UO3 x 2H2O). Citric acid formulations, consisting of oxalic acid-hydrogen peroxidecitric acid (OPC) or citric acid-hydrogen peroxidecitric acid (CPC), were used to remove uranium from the coupons. The efficiency of uranium removal varied from 68% to 94% depending on the extent of corrosion, the association of uranium with the iron oxide matrix, and the accessibility of the occluded contaminant. Decontaminated coupons clearly showed evidence of the extensive removal of rust and uranium. The waste solutions containing uranium and iron from decontamination by OPC and CPC were treated first by subjecting them to biodegradation followed by photodegradation. Biodegradation of a CPC solution by Pseudomonas fluorescens resulted in the degradation of the citric acid with concomitant precipitation of Fe (>96%), whereas U that remained in solution was recovered (>99%) by photodegradation as schoepite. In contrast, in an OPC solution citric acid was biodegraded but not oxalic acid, and both Fe and U remained in solution. Photodegradation of this OPC solution resulted in the precipitation of iron as ferrihydrite and uranium as uranyl hydroxide.