Peter Maxfield

Association of the Anxiogenic and Alerting Effects of Caffeine with ADORA2A and ADORA1 Polymorphisms and Habitual Level of Caffeine Consumption

Caffeine, a widely consumed adenosine A1 and A2A receptor antagonist, is valued as a psychostimulant, but it is also anxiogenic. An association between a variant within the ADORA2A gene (rs5751876) and caffeine-induced anxiety has been reported for individuals who habitually consume little caffeine. This study investigated whether this single nucleotide polymorphism (SNP) might also affect habitual caffeine intake, and whether habitual intake might moderate the anxiogenic effect of caffeine. Participants were 162 non-/low (NL) and 217 medium/high (MH) caffeine consumers. In a randomized, double-blind, parallel groups design they rated anxiety, alertness, and headache before and after 100 mg caffeine and again after another 150 mg caffeine given 90 min later, or after placebo on both occasions. Caffeine intake was prohibited for 16 h before the first dose of caffeine/placebo. Results showed greater susceptibility to caffeine-induced anxiety, but not lower habitual caffeine intake (indeed coffee intake was higher), in the rs5751876 TT genotype group, and a reduced anxiety response in MH vs NL participants irrespective of genotype. Apart from the almost completely linked ADORA2A SNP rs3761422, no other of eight ADORA2A and seven ADORA1 SNPs studied were found to be clearly associated with effects of caffeine on anxiety, alertness, or headache. Placebo administration in MH participants decreased alertness and increased headache. Caffeine did not increase alertness in NL participants. With frequent consumption, substantial tolerance develops to the anxiogenic effect of caffeine, even in genetically susceptible individuals, but no net benefit for alertness is gained, as caffeine abstinence reduces alertness and consumption merely returns it to baseline.

Acute impact of agriculture on high affinity methanotrophic bacterial populations

Exposure of mineral soils to atmospherically relevant concentrations of (13)CH(4) (2 ppmv) followed by (13)C-phospholipid fatty acid stable isotope probing allows assessment of the high-affinity methanotrophic bacterial sink in hitherto unattainable detail. Utilizing this approach, inorganic fertilizer-treated soils from a long-term agricultural experiment were shown to display dramatic reduction, by > 70%, of the methanotrophic bacterial cell numbers. Reduction in the methane sink capacity of the soils was slightly lower than the directly observed reduction in methanotrophic bacterial counts, indicating that the inhibitory effects on high-affinity methanotrophic bacteria are not fully expressed through CH(4) oxidation rates. The results emphasize the need to rigorously assess commonly applied agricultural practices with respect to their unseen negative impacts on soil microbial diversity in relation to terrestrial sinks for atmospheric trace gases.

Stable isotope switching (SIS): a new stable isotope probing (SIP) approach to determine carbon flow in the soil food web and dynamics in organic matter pools

RATIONALE Recent advances in stable isotope probing (SIP) have allowed direct linkage of microbial population structure and function. This paper details a new development of SIP, Stable Isotope Switching (SIS), which allows the simultaneous assessment of carbon (C) uptake, turnover and decay, and the elucidation of soil food webs within complex soils or sedimentary matrices. METHODS SIS utilises a stable isotope labelling approach whereby the (13)C-labelled substrate is switched part way through the incubation to a natural abundance substrate. A (13)CH(4) SIS study of landfill cover soils from Odcombe (Somerset, UK) was conducted. Carbon assimilation and dissimilation processes were monitored through bulk elemental analysis isotope ratio mass spectrometry and compound-specific gas chromatography/combustion/isotope ratio mass spectrometry, targeting a wide range of biomolecular components including: lipids, proteins and carbohydrates. RESULTS Carbon assimilation by primary consumers (methanotrophs) and sequential assimilation into secondary (Gram-negative and -positive bacteria) and tertiary consumers (Eukaryotes) was observed. Up to 45% of the bacterial membrane lipid C was determined to be directly derived from CH(4) and at the conclusion of the experiment ca. 50% of the bulk soil C derived directly from CH(4) was retained within the soil. CONCLUSIONS This is the first estimate of soil organic carbon derived from CH(4) and it is comparable with levels observed in lakes that have high levels of benthic methanogenesis. SIS opens the way for a new generation of SIP studies aimed at elucidating total C dynamics (incorporation, turnover and decay) at the molecular level in a wide range of complex environmental and biological matrices.

Archaeol: An Indicator of Methanogenesis in Water-Saturated Soils

Oxic soils typically are a sink for methane due to the presence of high-affinity methanotrophic Bacteria capable of oxidising methane. However, soils experiencing water saturation are able to host significant methanogenic archaeal communities, potentially affecting the capacity of the soil to act as a methane sink. In order to provide insight into methanogenic populations in such soils, the distribution of archaeol in free and conjugated forms was investigated as an indicator of fossilised and living methanogenic biomass using gas chromatography-mass spectrometry with selected ion monitoring. Of three soils studied, only one organic matter-rich site contained archaeol in quantifiable amounts. Assessment of the subsurface profile revealed a dominance of archaeol bound by glycosidic headgroups over phospholipids implying derivation from fossilised biomass. Moisture content, through control of organic carbon and anoxia, seemed to govern trends in methanogen biomass. Archaeol and crenarchaeol profiles differed, implying the former was not of thaumarcheotal origin. Based on these results, we propose the use of intact archaeol as a useful biomarker for methanogen biomass in soil and to track changes in moisture status and aeration related to climate change.

Stable isotope switching (SIS)

RATIONALE Recent advances in stable isotope probing (SIP) have allowed direct linkage of microbial population structure and function. This paper details a new development of SIP, Stable Isotope Switching (SIS), which allows the simultaneous assessment of carbon (C) uptake, turnover and decay, and the elucidation of soil food webs within complex soils or sedimentary matrices. METHODS SIS utilises a stable isotope labelling approach whereby the (13)C-labelled substrate is switched part way through the incubation to a natural abundance substrate. A (13)CH(4) SIS study of landfill cover soils from Odcombe (Somerset, UK) was conducted. Carbon assimilation and dissimilation processes were monitored through bulk elemental analysis isotope ratio mass spectrometry and compound-specific gas chromatography/combustion/isotope ratio mass spectrometry, targeting a wide range of biomolecular components including: lipids, proteins and carbohydrates. RESULTS Carbon assimilation by primary consumers (methanotrophs) and sequential assimilation into secondary (Gram-negative and -positive bacteria) and tertiary consumers (Eukaryotes) was observed. Up to 45% of the bacterial membrane lipid C was determined to be directly derived from CH(4) and at the conclusion of the experiment ca. 50% of the bulk soil C derived directly from CH(4) was retained within the soil. CONCLUSIONS This is the first estimate of soil organic carbon derived from CH(4) and it is comparable with levels observed in lakes that have high levels of benthic methanogenesis. SIS opens the way for a new generation of SIP studies aimed at elucidating total C dynamics (incorporation, turnover and decay) at the molecular level in a wide range of complex environmental and biological matrices.

Stable isotope switching (SIS): a new stable isotope probing (SIP) approach to determine carbon flow in the soil food web and dynamics in organic matter pools: 13C Stable isotope switching (SIS)

RATIONALE Recent advances in stable isotope probing (SIP) have allowed direct linkage of microbial population structure and function. This paper details a new development of SIP, Stable Isotope Switching (SIS), which allows the simultaneous assessment of carbon (C) uptake, turnover and decay, and the elucidation of soil food webs within complex soils or sedimentary matrices. METHODS SIS utilises a stable isotope labelling approach whereby the (13)C-labelled substrate is switched part way through the incubation to a natural abundance substrate. A (13)CH(4) SIS study of landfill cover soils from Odcombe (Somerset, UK) was conducted. Carbon assimilation and dissimilation processes were monitored through bulk elemental analysis isotope ratio mass spectrometry and compound-specific gas chromatography/combustion/isotope ratio mass spectrometry, targeting a wide range of biomolecular components including: lipids, proteins and carbohydrates. RESULTS Carbon assimilation by primary consumers (methanotrophs) and sequential assimilation into secondary (Gram-negative and -positive bacteria) and tertiary consumers (Eukaryotes) was observed. Up to 45% of the bacterial membrane lipid C was determined to be directly derived from CH(4) and at the conclusion of the experiment ca. 50% of the bulk soil C derived directly from CH(4) was retained within the soil. CONCLUSIONS This is the first estimate of soil organic carbon derived from CH(4) and it is comparable with levels observed in lakes that have high levels of benthic methanogenesis. SIS opens the way for a new generation of SIP studies aimed at elucidating total C dynamics (incorporation, turnover and decay) at the molecular level in a wide range of complex environmental and biological matrices.