Methane as a resource: can the methanotrophs add value?

Methane is an abundant gas used in energy recovery systems, heating, and transport. Methanotrophs are bacteria capable of using methane as their sole carbon source. Although intensively researched, the myriad of potential biotechnological applications of methanotrophic bacteria has not been comprehensively discussed in a single review. Methanotrophs can generate single-cell protein, biopolymers, components for nanotechnology applications (surface layers), soluble metabolites (methanol, formaldehyde, organic acids, and ectoine), lipids (biodiesel and health supplements), growth media, and vitamin B12 using methane as their carbon source. They may be genetically engineered to produce new compounds such as carotenoids or farnesene. Some enzymes (dehydrogenases, oxidase, and catalase) are valuable products with high conversion efficiencies and can generate methanol or sequester CO2 as formic acid ex vivo. Live cultures can be used for bioremediation, chemical transformation (propene to propylene oxide), wastewater denitrification, as components of biosensors, or possibly for directly generating electricity. This review demonstrates the potential for methanotrophs and their consortia to generate value while using methane as a carbon source. While there are notable challenges using a low solubility gas as a carbon source, the massive methane resource, and the potential cost savings while sequestering a greenhouse gas, keeps interest piqued in these unique bacteria.

Voriconazole therapeutic drug monitoring: results of a prematurely discontinued randomized multicenter trial.

BACKGROUND: Voriconazole (VOR) levels are highly variable, with potential implications to both efficacy and safety. We hypothesized that VOR therapeutic drug monitoring (TDM) will decrease the incidence of treatment failures and adverse events (AEs). METHODS: We initiated a prospective, randomized, non-blinded multicenter study to compare clinical outcomes in adult patients randomized to standard dosing (clinician-driven) vs. TDM (doses adjusted based on levels). VOR trough levels were obtained on day 5, 14, 28, and 42 (or at completion of drug; ± 3 days). Real-time dose adjustments were made to maintain a range between 1-5 µg/mL on the TDM-arm, while levels were assessed retrospectively in the standard-arm. Patient questionnaires were administered to assess subjective AEs. RESULTS: The study was discontinued prematurely, after 29 patients were enrolled. Seventeen (58.6%) patients experienced 38 AEs: visual changes (22/38, 57.9%), neurological symptoms (13/38, 34.2%), and liver abnormalities (3/38, 7.9%). VOR was discontinued in 7 (25%) patients because of an AE (4 standard-arm, 3 TDM-arm). VOR levels were frequently out of range in the standard-arm (8 tests >5 µg/mL; 9 tests <1 µg/mL). Three dose changes occurred in the TDM-arm for VOR levels <1 µg/mL. Levels decreased over time in the standard-arm, with mean VOR levels lower at end of therapy compared to TDM (1.3 vs. 4.6 µg/mL, P = 0.008). CONCLUSIONS: VOR TDM has become widespread clinical practice, based on known variability in drug levels, which impaired accrual in this study. Although comparative conclusions are limited, observations of variability and waning levels over time support TDM.

Pancreatic agenesis attributable to a single nucleotide deletion in the human IPF1 gene coding sequence.

The homeodomain protein IPF1 (also known as IDX1, STF1 and PDX1; see Methods) is critical for development of the pancreas in mice and is a key factor for the regulation of the insulin gene in the beta-cells of the endocrine pancreas. Targeted disruption of the Ipf1 gene encoding IPF1 in transgenic mice results in a failure of the pancreas to develop (pancreatic agenesis). Here, we report the identification of a single nucleotide deletion within codon 63 of the human IPF1 gene (13q12.1) in a patient with pancreatic agenesis. The patient is homozygous for the point deletion, whereas both parents are heterozygotes for the same mutation. The deletion was not found in 184 chromosomes from normal individuals, indicating that the mutation is unlikely to be a rare polymorphism. The point deletion causes a frame shift at the C-terminal border of the transactivation domain of IPF1 resulting in the translation of 59 novel codons before termination, aminoproximal to the homeodomain essential for DNA binding. Expression of mutant IPF1 in Cos-1 cells confirms the expression of a prematurely terminated truncated protein of 16 kD. Thus, the affected patient should have no functional IPF1 protein. Given the essential role of IPF1 in pancreas development, it is likely that this autosomal recessive mutation is the cause of the pancreatic agenesis phenotype in this patient. Thus, IPF1 appears to be a critical regulator of pancreas development in humans as well as mice.

Fate of pathogen indicators in a domestic blend of food waste and wastewater through a two-stage anaerobic digestion system.

Anaerobic digestion is a viable on-site treatment technology for rich organic waste streams such as food waste and blackwater. In contrast to large-scale municipal wastewater treatment plants which are typically located away from the community, the effluent from any type of on-site system is a potential pathogenic hazard because of the intimacy of the system to the community. The native concentrations of the pathogen indicators Escherichia coli, Clostridium perfringens and somatic coliphage were tracked for 30 days under stable operation (organic loading rate (OLR) = 1.8 kgCOD m(-3) day(-1), methane yield = 52% on a chemical oxygen demand (COD) basis) of a two-stage laboratory-scale digester treating a mixture of food waste and blackwater. E. coli numbers were reduced by a factor of 10(6.4) in the thermophilic stage, from 10(7.5±0.3) to 10(1.1±0.1) cfu 100 mL(-1), but regenerated by a factor of 10(4) in the mesophilic stage. Neither the thermophilic nor mesophilic stages had any significant impact on C. perfringens concentrations. Coliphage concentrations were reduced by a factor of 10(1.4) across the two stages. The study shows that anaerobic digestion only reduces pathogen counts marginally but that counts in effluent samples could be readily reduced to below detection limits by filtration through a 0.22 µm membrane, to investigate membrane filtration as a possible sanitation technique.

The World Rugby and International Rugby Players Contact Load Guidelines: From conception to implementation and the future.

Managing training load in rugby union is crucial for optimising performance and injury prevention. Contact training warrants attention because of higher overall injury and head impact risk, yet players must develop physical, technical, and mental skills to withstand the demands of the game. To help coaches manage contact loads in professional rugby, World Rugby and International Rugby Players convened an expert working group. They conducted a global survey with players to develop contact load guidelines. This commentary aims to describe the contact load guidelines and their implementation, and identify areas where future work is needed to support their evolution.

Syncoilin modulates peripherin filament networks and is necessary for large-calibre motor neurons.

Syncoilin is an atypical type III intermediate filament (IF) protein, which is expressed in muscle and is associated with the dystrophin-associated protein complex. Here, we show that syncoilin is expressed in both the central and peripheral nervous systems. Isoform Sync1 is dominant in the brain, but isoform Sync2 is dominant in the spinal cord and sciatic nerve. Peripherin is a type III IF protein that has been shown to colocalise and interact with syncoilin. Our analyses suggest that syncoilin might function to modulate formation of peripherin filament networks through binding to peripherin isoforms. Peripherin is associated with the disease amyotrophic lateral sclerosis (ALS), thus establishing a link between syncoilin and ALS. A neuronal analysis of the syncoilin-null mouse (Sync(-/-)) revealed a reduced ability in accelerating treadmill and rotarod tests. This phenotype might be attributable to the impaired function of extensor digitorum longus muscle and type IIb fibres caused by a shift from large- to small-calibre motor axons in the ventral root.

Current status of inverse agonism at serotonin2A (5-HT2A) and 5-HT2C receptors.

Contemporary receptor theory was developed to account for the existence of constitutive activity, as defined by the presence of receptor signaling in the absence of any ligand. Thus, ligands acting at a constitutively active receptor, can act as agonists, antagonists, and inverse agonists. In vitro studies have also revealed the complexity of ligand/receptor interactions including agonist-directed stimulus trafficking, a finding that has led to multi-active state models of receptor function. Studies with a variety of cell types have established that the serotonin 5-HT(2A) and 5-HT(2C) receptors also demonstrate constitutive activity and inverse agonism. However, until recently, there has been no evidence to suggest that these receptors also demonstrate constitutive activity and hence reveal inverse agonist properties of ligands in vivo. This paper describes our current knowledge of constitutive activity in vitro and then examines the evidence for constitutive activity in vivo. Both the serotonin 5-HT(2A) and 5-HT(2C) receptors are involved in a number of physiological and behavioral functions and are the targets for treatment of schizophrenia, anxiety, weight control, Parkinsonism, and other disorders. The existence of constitutive activity at these receptors in vivo, along with the possibility of inverse agonism, provides new avenues for drug development.

Influence of molecular oxide cages on metal carbonyls.

A series of DFT calculations have been performed on mononuclear d(6) metal tricarbonyls supported on molecular oxide cages. The molecular cages were chosen both as models for phosphate, silicate, and aluminosilicate surfaces and because experimental data is available for a few of these molecular complexes. By systematically varying the nature of the oxide surface, qualitative estimates of metal carbonyl geometry, relative metal-CO bond strengths, and predictions of the shifts in C identical withO stretching frequencies upon changes in oxide basicity have been determined for a range of transition metals. Although most of the calculated trends correlate with expectations, additional insights into some of the bonding characteristics of these systems were obtained.

Child oral health concerns amongst parents and primary care givers in a Sure Start local programme.

AIM: To conduct an oral health promotion needs assessment amongst parents and primary care givers of pre-school children in a South East London Sure Start Local Programme (SSLP). OBJECTIVE: To explore the oral health concerns and oral health literacy with regard to children's oral health amongst parents and primary care givers in a South East London SSLP. DESIGN: A qualitative study using four in-depth focus groups with a purposive sample of 20 participants. Data were analysed using the framework method. RESULTS: The SSLP was identified as an important source of information, support and social interaction for participants. Participants rated the informal networks of the programme as equally authoritative as other formal sources of information. Oral health concerns included: introducing healthy eating, establishing tooth brushing, teething and access to dental care. While participants had adequate knowledge of how to prevent oral disease they cited many barriers to acting on their knowledge which included: parents' tiredness, lack of confidence in parenting skills, confusing information, widespread availability of sugary foods and drinks, and lack of local child friendly dentists. Parenting skills and the social support provided by the SSLP appeared to be integral to the introduction of positive oral health behaviours. CONCLUSIONS: SSLPs were seen as a trusted source of support and information for carers of pre-school children. Integration of oral health promotion into SSLPs has the potential to tap into early interventions which tackle the wider support needs of carers of pre-school children while also supporting the development of positive oral health behaviours.

Syncoilin isoform organization and differential expression in murine striated muscle.

Syncoilin is a 64kDa intermediate filament (IF) protein expressed in myocytes at the sarcolemma, perinucleus, myotendenous and neuromuscular junctions. Here we present a revised domain projection and structural analysis for the original isoform (sync-1) and introduce two novel syncoilin isoforms (sync-2 and sync-3) generated by exon splicing. On the basis of consensus identity we propose that syncoilin be reclassified as a type III IF protein. All three syncoilin isoforms lack a L1 domain, a significant departure from standard IF rod domain projections that is likely to impact significantly on their biological function. Our analyses indicate that syncoilin is unlikely to form classical intermediate filament structures by itself, and that the significant difference in C-terminal structure between the three isoforms indicates that they may play divergent roles in myocytes. We show that despite lacking an apparent structural role in striated muscle, syncoilin isoforms are differentially and strongly upregulated in response to cardiotoxin induced regeneration and denervation induced atrophy in the C57BL/6 mouse, possibly suggesting an atypical role for syncoilin in muscle.

Excess helium-4 in teggau lake: possibilities for a uranium ore body.

Excess (4)He (more than five times the solubility) has been measured in Teggau Lake in northwestern Ontario. A model suggests that an adjacent mass of greater than 10(4) kilograms of uranium is responsible for the observed (4)He excess. The area is favorable for pegmatitic uranium deposits, and the release of trapped (4)He from uraninite dikes larger than 30 cubic meters (1 percent U(3)O(8)) provides the best explanation for the excess (4)He in the lake.

Spontaneous fission-neutron fission xenon: a new technique for dating geological events.

A method for dating geological samples which uses fission product xenon in a manner similar to the use of radiogenic argon in the 40Ar-39Ar technique has been developed. The results of stepwise heating experiments for a zircon from the Ahaggar region in the Sahara are compared to the geochronology determined by the rubidiumstrontium, uranium-thorium-lead, and potassium-argon dating methods.

Strontium-90: effects of chronic ingestion on farrowing performance of miniature swine.

In experiments involving the ingestion of strontium-90 by nearly 800 female miniature swine and extending over three generations, no significant differences in litter size, percentage of stillborn, or birth weight were observed between controls and animals ingesting up to 625 microcuries of strontium-90 per day. At 625 microcuries per day, these animals were ingesting more than a million times the peak value of strontium-90 ever reported in the American diet. Animals on 3100 microcuries per day did not survive the gestation period. From these studies, it is evident that feeding levels of strontium-90 high enough to affect fetal or neonatal mortality in this species will not permit maternal survival long enough for the bearing of young.

Ceratoscopelus maderensis: peculiar sound-scattering layer identified with this myctophid fish.

A sound-scattering layer, composed of discrete hyperbolic echosequences and apparently restricted to the Slope Water region of the western North Atlantic, has been identified from the Deep Submergence Research Vehicle Alvin with schools of the meyctophid fish Ceratoscopelus maderensis. By diving into the layer and using Alvin's echo-ranging sonar, we approached and visually identified the sound scatterers. The number of echo sequences observed with the surface echo-soutnder (1/23.76 x 10(5) cubic meters of water) checked roughly with the number of sonar targets observed from the submarine (1/7.45 x 105 cubic meters). The fish schools appeared to be 5 to 10 meters thick, 10 to 100 meters in diameter, and on centers 100 to 200 meters apart. Density within schools was estimated at 10 to 15 fish per cubic meter.

The effect of biomass density on cellulose solubilisation rates.

The aim of this work was to compare the impact of inoculation density on the rate of cellulose hydrolysis by a rumen derived culture with that of a microbial enrichment from an organic waste anaerobic digester. The results showed a linear relationship between the mass of biomass at the start of the first order degradation phase (Xo) and the first order hydrolysis rate (r) for both rumen inoculated and leachate inoculated cellulose digestions and that the slopes of these relationships were not distinguishable. This suggested that differences in the microbial community, media and other environmental factors had a lesser impact on the hydrolysis rate compared to the effect of the number of cells in the system. This could be of great importance to industrial applications of anaerobic digestion technologies as it suggested that if cells densities in the waste treatment digesters could be boosted to match those seen in the rumen, then the rates of the cellulose hydrolysis would rise.

Digestion of waste bananas to generate energy in Australia.

This paper presents results from laboratory studies to measure the methane yield and rate of digestion of reject bananas. These parameters were determined in experiments that took into account the likely configuration of a full-scale plant in the banana growing region of north Queensland. The digestion was conducted in a 200-l reactor using fed-batch operation, relying entirely on the natural microbial consortia on the reject bananas to avoid reliance on external inocula such as sludge, an undesirable material around food packaging facilities. An enrichment culture was first established in a highly buffered 200-l batch digestion unit. The fed-batch digester was then started by exchanging leachate with the mature batch reactor. Under loading conditions of 0.6 kg VS m(-3)d(-1) over 70 days where the average working volume was 160 l, the digester produced 398+/-20 l CH4 kg VS(-1). Increasing the loading rate to 1.6 kg VS m(-3)d(-1) resulted in a reduced methane yield of 210 l CH4 kg VS(-1) over 23 days of operation, with a concomitant accumulation of banana waste in the digester. The leachate at the end of digestion contained over 4000 mg l(-1)K, 200 mg l(-1) N and 75 mg l(-1), levels that exceed acceptable limits for general agricultural irrigation.

Measurement and quantification of sessile and planktonic microbial populations during the anaerobic digestion of cellulose.

Crystalline cellulose was anaerobically degraded using a leachate inoculum derived from simulated municipal solid waste. Bicinchoninic Acid (BCA) protein assays were used to measure the distribution of biomass during cellulose degradation, including the planktonic and sessile biomass fractions. A comparison of sessile and planktonic microbial growth indicated that the microbial growth was dominated by the planktonic fraction with the biofilms accounting for approximately 25% of the population. Additional biomass measurements were conducted to test the reliability of the BCA protein assays. Total microbial growth was inferred from the accumulation and depletion of ammonia nitrogen measured using flow injection analysis. The planktonic biomass was estimated from direct cell counts using light microscopy and the sessile biomass was estimated by analysing the nitrogen content of the separated and washed cellulose pellet. Regression analysis showed good correlations between the measurement pairs representing the total biomass (R2=0.90), planktonic biomass (R2=0.97) and sessile biomass (R2=0.85), supporting the use of protein assays as an indicator of microbial growth in mixed culture environments.

Understanding the motivation: a qualitative study of dental students' choice of professional career.

BACKGROUND: Given the changing nature of the dental workforce, and the need to retain the services of future members, it is important to understand why current dental students perceive that they were motivated to study dentistry. Qualitative research provides the opportunity to explore the underlying issues in addition to informing subsequent quantitative research. The objectives of this research were to investigate final-year dental students' motivation for studying dentistry and how they perceive this has been modified during their undergraduate degree programme. METHODS: Purposive sampling of a representative group of 35 final-year dental students at King's College London Dental Institute to participate in audio-taped focus groups. Qualitative data were analysed using Framework Methodology. RESULTS: The findings suggest a strong emphasis on having a career, providing 'professional status', 'financial benefits', 'job security, flexibility and independence' and 'good quality of life'. Students reported being attracted by features of the job, supported to a greater or lesser extent by personal experience, family and friends. It appears however that students' initial motivation is being tempered by their experiences during their undergraduate degree programme, in particular, the 'responsibilities of an intensive professional education', their 'mounting student debt' and the perception of 'feeling undervalued'. This perception related to dentistry in general and National Health Service dentistry in particular, being undervalued, by government, patients, the public and members of the dental profession. CONCLUSIONS: Students' vision of a 'contained professional career' within health care, providing status and financial benefits, appears to have influenced their choice of dentistry. Pressures relating to student life and policy changes are perceived as impacting on key components of professional life, particularly status in the social and economic order. The implications for educators, professional leaders and policy makers are explored.

The uptake of anaerobic digestion for the organic fraction of municipal solid waste - Push versus pull factors.

There are thousands of anaerobic digestion facilities worldwide applied to agricultural waste, energy crops and industrial food processing wastes. Yet, centralised anaerobic digestion for the organic fraction of Municipal Solid Waste (OFMSW) is almost exclusively applied in Europe and California where diversion of organics from landfill is enforced by legislation. Even in the EU however, only 5% of OFMSW is currently digested, with most organic waste incinerated. Municipalities elsewhere are reluctant to switch to biological treatment, even when made financially preferable through mechanisms such as landfill taxes. The uptake of OFMSW anaerobic digestion around the world and the legislative and economic drivers where this has occurred are reviewed. The prime reason for lack of uptake in regions with ample economic drivers is a lack of experience of municipalities and environmental regulators in regulating both the facilities and the disposal of treated organic material to land.