The metabolic rate of the biosphere and its components.

We assessed the relationship between rates of biological energy utilization and the biomass sustained by that energy utilization, at both the organism and biosphere level. We compiled a dataset comprising >10,000 basal, field, and maximum metabolic rate measurements made on >2,900 individual species, and, in parallel, we quantified rates of energy utilization, on a biomass-normalized basis, by the global biosphere and by its major marine and terrestrial components. The organism-level data, which are dominated by animal species, have a geometric mean among basal metabolic rates of 0.012 W (g C)-1 and an overall range of more than six orders of magnitude. The biosphere as a whole uses energy at an average rate of 0.005 W (g C)-1 but exhibits a five order of magnitude range among its components, from 0.00002 W (g C)-1 for global marine subsurface sediments to 2.3 W (g C)-1 for global marine primary producers. While the average is set primarily by plants and microorganisms, and by the impact of humanity upon those populations, the extremes reflect systems populated almost exclusively by microbes. Mass-normalized energy utilization rates correlate strongly with rates of biomass carbon turnover. Based on our estimates of energy utilization rates in the biosphere, this correlation predicts global mean biomass carbon turnover rates of ~2.3 y-1 for terrestrial soil biota, ~8.5 y-1 for marine water column biota, and ~1.0 y-1 and ~0.01 y-1 for marine sediment biota in the 0 to 0.1 m and >0.1 m depth intervals, respectively.

A dynamic microbial sulfur cycle in a serpentinizing continental ophiolite.

Serpentinization is the hydration and oxidation of ultramafic rock, which occurs as oceanic lithosphere is emplaced onto continental margins (ophiolites), and along the seafloor as faulting exposes this mantle-derived material to circulating hydrothermal fluids. This process leads to distinctive fluid chemistries as molecular hydrogen (H2 ) and hydroxyl ions (OH- ) are produced and reduced carbon compounds are mobilized. Serpentinizing ophiolites also serve as a vector to transport sulfur compounds from the seafloor onto the continents. We investigated hyperalkaline, sulfur-rich, brackish groundwater in a serpentinizing continental ophiolite to elucidate the role of sulfur compounds in fuelling in situ microbial activities. Here we illustrate that key sulfur-cycling taxa, including Dethiobacter, Desulfitispora and 'Desulforudis', persist throughout this extreme environment. Biologically catalysed redox reactions involving sulfate, sulfide and intermediate sulfur compounds are thermodynamically favourable in the groundwater, which indicates they may be vital to sustaining life in these characteristically oxidant- and energy-limited systems. Furthermore, metagenomic and metatranscriptomic analyses reveal a complex network involving sulfate reduction, sulfide oxidation and thiosulfate reactions. Our findings highlight the importance of the complete inorganic sulfur cycle in serpentinizing fluids and suggest sulfur biogeochemistry provides a key link between terrestrial serpentinizing ecosystems and their submarine heritage.

Probing the geological source and biological fate of hydrogen in Yellowstone hot springs.

Hydrogen (H2 ) is enriched in hot springs and can support microbial primary production. Using a series of geochemical proxies, a model to describe variable H2 concentrations in Yellowstone National Park (YNP) hot springs is presented. Interaction between water and crustal iron minerals yields H2 that partition into the vapour phase during decompressional boiling of ascending hydrothermal fluids. Variable vapour input leads to differences in H2 concentration among springs. Analysis of 50 metagenomes from a variety of YNP springs reveals that genes encoding oxidative hydrogenases are enriched in communities inhabiting springs sourced with vapour-phase gas. Three springs in the Smokejumper (SJ) area of YNP that are sourced with vapour-phase gas and with the most H2 in YNP were examined to determine the fate of H2 . SJ3 had the most H2 , the most 16S rRNA gene templates and the greatest abundance of culturable hydrogenotrophic and autotrophic cells of the three springs. Metagenomics and transcriptomics of SJ3 reveal a diverse community comprised of abundant populations expressing genes involved in H2 oxidation and carbon dioxide fixation. These observations suggest a link between geologic processes that generate and source H2 to hot springs and the distribution of organisms that use H2 to generate energy.

Alcohol Use and Drinking Motives Among Suddenly Bereaved College Students.

Objective: Thousands of Americans grieve the sudden death of a loved one each year. These sudden deaths may be the result of violent deaths, as in the cases of homicide, suicide, or fatal accidents, or may occur as the result of natural causes such as heart attacks or strokes. Sudden loss survivors often experience negative mental health outcomes such as depression, prolonged grief disorder, and posttraumatic stress disorder. Survivors may also misuse alcohol for varied reasons after these losses, which can put them at risk for alcohol-related consequences. Thus, the purpose of this study was to explore associations between psychological distress and alcohol-related outcomes among young adults with a history of sudden loss. Methods: A sample of 659 young adults completed a series of self-report measures assessing loss history, psychosocial distress, perceived alcohol-related problems, and drinking motives. Results: Results showed that survivors of sudden, violent losses reported higher rates of past 30-day alcohol use than those who had lost a loved one to a sudden, natural loss or those who reported no loss history. Although there were no statistically significant differences in drinking motives between violent or natural loss survivors, more severe depression symptoms among bereaved individuals were associated with drinking to cope and to conform. Participants experiencing prolonged grief symptoms were also more likely to report drinking to conform. Conclusions: Bereavement-related distress may influence drinking motives among young adults with a history of sudden loss.

Comparison of Bioelectrical Impedance Analysis and Dual-Energy X-Ray Absorptiometry for Estimating Bone Mineral Content.

The purpose of this study was to validate single-frequency hand-to-foot bioelectrical impedance analysis (HFBIA) for estimating bone mineral content (BMC) using dual-energy X-ray absorptiometry as the criterion measure in healthy men and women aged 18-40 years. A total of 80 men and women participated in this study. BMC was estimated on the same day using HFBIA and dual-energy X-ray absorptiometry. The HFBIA device provided higher mean BMC values in men and the entire sample, but not in women. A smaller standard error of estimate was observed in women (0.20, corresponding to 8% of the mean reference BMC values) compared with men (0.39, corresponding to 12% of the mean reference BMC values) and the combined sample (0.31). HFBIA provided a smaller constant error and individual estimation error indicated by the 95% limits of agreement in women (-0.05 ± 0.39) compared with men (-0.16 ± 0.78) and the entire sample (-0.10 ± 0.63). In conclusion, although BMC values were found to be more accurate in women, HFBIA overestimated BMC compared with dual-energy X-ray absorptiometry, especially in individuals with lower values. Given these results, using HFBIA to measure BMC would be inappropriate for diagnostic purposes.

Connexin Hemichannels: Methods for Dye Uptake and Leakage.

It is now clear that connexin-based, gap junction "hemichannels" in an undocked state are capable of opening and connecting cytoplasm to the extracellular milieu. Varied studies also suggest that such channel activity plays a vital role in diverse cell processes and abnormal hemichannel activity contributes to pathogenesis. To pursue fundamental questions in this area, investigators require methods for studying hemichannel permeability and dynamics that are quantitative, sensitive, versatile, and available to most cellular and molecular laboratories. Here we first provide a theoretical background for this work, including the role of cellular membrane potentials. We then describe in detail our computer-assisted methods for both dye uptake and leakage along with illustrative results from different cell systems. A key feature of our protocol is the inclusion of a mechanical stimulation step. We describe dye uptake, interpreted as connexin dependent, that is shown to be enhanced with reduced extracellular Ca2+, mechanically responsive, inhibited by TPA, inhibited by EL186 antibodies for Cx43 and sustained for more than 15 min following mechanical stimulation. We describe dye leakage that displays these same properties, with estimates of hemichannel numbers per cell being derived from leakage rates. We also describe dye uptake that is shown to be unaffected by a reduction in external Ca2+, insensitive to EL186 antibodies and relatively short-lived following mechanical stimulation; this uptake may occur via pannexin 1 channels expressed in the cells studied here. It is unlikely that cell damage plays a significant role in dye uptake following mechanical stimulation, given compelling results from various control experiments.

Carbon source preference in chemosynthetic hot spring communities.

Rates of dissolved inorganic carbon (DIC), formate, and acetate mineralization and/or assimilation were determined in 13 high-temperature (>73 °C) hot springs in Yellowstone National Park (YNP), Wyoming, in order to evaluate the relative importance of these substrates in supporting microbial metabolism. While 9 of the hot spring communities exhibited rates of DIC assimilation that were greater than those of formate and acetate assimilation, 2 exhibited rates of formate and/or acetate assimilation that exceeded those of DIC assimilation. Overall rates of DIC, formate, and acetate mineralization and assimilation were positively correlated with spring pH but showed little correlation with temperature. Communities sampled from hot springs with similar geochemistries generally exhibited similar rates of substrate transformation, as well as similar community compositions, as revealed by 16S rRNA gene-tagged sequencing. Amendment of microcosms with small (micromolar) amounts of formate suppressed DIC assimilation in short-term (<45-min) incubations, despite the presence of native DIC concentrations that exceeded those of added formate by 2 to 3 orders of magnitude. The concentration of added formate required to suppress DIC assimilation was similar to the affinity constant (K(m)) for formate transformation, as determined by community kinetic assays. These results suggest that dominant chemoautotrophs in high-temperature communities are facultatively autotrophic or mixotrophic, are adapted to fluctuating nutrient availabilities, and are capable of taking advantage of energy-rich organic substrates when they become available.

Threshold in breast compression reduction for full-field digital mammography and digital breast tomosynthesis.

INTRODUCTION: Breast compression is essential in mammography to improve image quality and reduce radiation dose. However, it can cause discomfort or even pain in women which could discourage them from attending future mammography examinations. Therefore, this study aims to explore the maximum reduction in breast compression in full-field digital mammography (FFDM) and digital breast tomosynthesis (DBT) that is achievable without impacting on image quality and dose. METHODS: Ten compression force (CF) levels (20N-110N, with 10N intervals) were assessed on Siemens MAMMOMAT Inspiration with Nuclear Associates 18-228 phantom. Imaging was carried out in craniocaudal projection using Automatic Exposure Control at 28 kVp with a Tungsten/Rhodium anode/filter combination, and at 50° sweep angle for DBT. Using ImageJ software, image quality of the acquired mammograms and central tomosynthesis slices were examined based on mass conspicuity (MC) and microcalcification conspicuity (MicroC). Entrance skin dose (ESD) and mean glandular dose (MGD) were recorded from Digital Imaging and Communication in Medicine image header. Linear regression was performed to examine the relationship between CF with ESD, MGD, MC and MicroC. Differences in image quality and radiation dose were assessed with one-way analysis of variance and Kruskal-Wallis H test. RESULTS: Significant correlations were noted between CF with ESD and MicroC for FFDM and DBT, with DBT also demonstrating associations with MGD and MC. No significant differences were observed for ESD, MGD, MC and MicroC when CF was reduced to 40N and 80N in FFDM and DBT respectively. CONCLUSION: This study demonstrated that CF can be reduced as low as 40N and 80N in FFDM and DBT respectively, without significant impact on image quality and radiation dose. IMPLICATIONS FOR PRACTICE: Reduced mammographic compression may reduce discomfort or pain in women, which may improve attendance rate in breast screening programmes. Findings from this study will provide reference for future work examining breast compression in mammography.

Competition for inorganic carbon between oxygenic and anoxygenic phototrophs in a hypersaline microbial mat, Guerrero Negro, Mexico.

While most oxygenic phototrophs harvest light only in the visible range (400-700 nm, VIS), anoxygenic phototrophs can harvest near infrared light (> 700 nm, NIR). To study interactions between the photosynthetic guilds we used microsensors to measure oxygen and gross oxygenic photosynthesis (gOP) in a hypersaline microbial mat under full (VIS + NIR) and VIS illumination. Under normal dissolved inorganic carbon (DIC) concentrations (2 mM), volumetric rates of gOP were reduced up to 65% and areal rates by 16-31% at full compared with VIS illumination. This effect was enhanced (reduction up to 100% in volumetric, 50% in areal rates of gOP) when DIC was lowered to 1 mM, but diminished at 10 mM DIC or lowered pH. In conclusion, under full-light illumination anoxygenic phototrophs are able to reduce the activity of oxygenic phototrophs by efficiently competing for inorganic carbon within the highly oxygenated layer. Anoxygenic photosynthesis, calculated from the difference in gOP under full and VIS illumination, represented between 10% and 40% of the C-fixation. The DIC depletion in the euphotic zone as well as the significant C-fixation by anoxygenic phototrophs in the oxic layer influences the carbon isotopic composition of the mat, which needs to be taken into account when interpreting isotopic biosignals in geological records.

Modeled energetics of bacterial communities in ancient subzero brines.

Cryopeg brines are isolated volumes of hypersaline water in subzero permafrost. The cryopeg system at Utqiagvik, Alaska, is estimated to date back to 40 ka BP or earlier, a remnant of a late Pleistocene Ocean. Surprisingly, the cryopeg brines contain high concentrations of organic carbon, including extracellular polysaccharides, and high densities of bacteria. How can these physiologically extreme, old, and geologically isolated systems support such an ecosystem? This study addresses this question by examining the energetics of the Utqiagvik cryopeg brine ecosystem. Using literature-derived assumptions and new measurements on archived borehole materials, we first estimated the quantity of organic carbon when the system formed. We then considered two bacterial growth trajectories to calculate the lower and upper bounds of the cell-specific metabolic rate of these communities. These bounds represent the first community estimates of metabolic rate in a subzero hypersaline environment. To assess the plausibility of the different growth trajectories, we developed a model of the organic carbon cycle and applied it to three borehole scenarios. We also used dissolved inorganic carbon and nitrogen measurements to independently estimate the metabolic rate. The model reconstructs the growth trajectory of the microbial community and predicts the present-day cell density and organic carbon content. Model input included measured rates of the in-situ enzymatic conversion of particulate to dissolved organic carbon under subzero brine conditions. A sensitivity analysis of model parameters was performed, revealing an interplay between growth rate, cell-specific metabolic rate, and extracellular enzyme activity. This approach allowed us to identify plausible growth trajectories consistent with the observed bacterial densities in the cryopeg brines. We found that the cell-specific metabolic rate in this system is relatively high compared to marine sediments. We attribute this finding to the need to invest energy in the production of extracellular enzymes, for generating bioavailable carbon from particulate organic carbon, and the production of extracellular polysaccharides for cryoprotection and osmoprotection. These results may be relevant to other isolated systems in the polar regions of Earth and to possible ice-bound brines on worlds such as Europa, Enceladus, and Mars.

Corrigendum: Modeled energetics of bacterial communities in ancient subzero brines.

[This corrects the article DOI: 10.3389/fmicb.2023.1206641.].

Phosphate availability and implications for life on ocean worlds.

Several moons in the outer solar system host liquid water oceans. A key next step in assessing the habitability of these ocean worlds is to determine whether life's elemental and energy requirements are also met. Phosphorus is required by all known life and is often limited to biological productivity in Earth's oceans. This raises the possibility that its availability may limit the abundance or productivity of Earth-like life on ocean worlds. To address this potential problem, here we calculate the equilibrium dissolved phosphate concentrations associated with the reaction of water and rocks-a key driver of ocean chemical evolution-across a broad range of compositional inputs and reaction conditions. Equilibrium dissolved phosphate concentrations range from 10-11 to 10-1 mol/kg across the full range of carbonaceous chondrite compositions and reaction conditions considered, but are generally > 10-5 mol/kg for most plausible scenarios. Relative to the phosphate requirements and uptake kinetics of microorganisms in Earth's oceans, such concentrations would be sufficient to support initially rapid cell growth and construction of global ocean cell populations larger than those observed in Earth's deep oceans.

Receptor chimeras demonstrate that the C-terminal domain of the human cytomegalovirus US27 gene product is necessary and sufficient for intracellular receptor localization.

BACKGROUND: Human cytomegalovirus (HCMV) is ubiquitous in the population but generally causes only mild or asymptomatic infection except in immune suppressed individuals. HCMV employs numerous strategies for manipulating infected cells, including mimicry of G-protein coupled receptors (GPCRs). The HCMV US27 gene product is a putative GPCR, yet no ligand or signaling has been identified for this receptor. In the present study, immunofluorescence microscopy was used to examine the cellular distribution of wild type US27, as well as US27 deletion mutants and chimeric receptors. RESULTS: In transiently transfected cells, wild type US27 was found primarily in intracellular compartments, in striking contrast to the cell surface distribution seen for the human cellular chemokine receptor CXCR3. When the N-terminal extracellular domains of the two receptors were swapped, no change in protein localization was observed. However, swapping of the C-terminal intracellular domains resulted in a significant change in receptor distribution. A chimera that contained US27 fused to the C-terminal intracellular tail of CXCR3 exhibited surface distribution similar to that of wild-type CXCR3. When the C-terminal domain of US27 was fused to CXCR3, this chimeric receptor (CXCR3/US27-CT) was found in the same intracellular pattern as wild-type US27. In addition, a US27 mutant lacking the C-terminus (US27ΔCT) failed to accumulate inside the cell and exhibited cell surface distribution. Co-localization with organelle-specific markers revealed that wild-type US27 was found predominantly in the Golgi apparatus and in endosomal compartments, whereas the US27/CXCR3-CT chimera, US27ΔCT and US27Δ348 mutants were not localized to endosomal compartments. CONCLUSIONS: The results indicate that the C-terminal domain of the HCMV US27 protein, which contains a di-leucine endocytic sorting motif, is both necessary and sufficient for intracellular localization, which may also help explain why no cellular ligands have yet been identified for this viral receptor.

Probing Internal Assumptions of the Revised Bloom's Taxonomy.

Bloom's taxonomy is a classification of learning objectives originally developed for general educational purposes. The taxonomy was revised to expand beyond cognitive processes and to include knowledge types as an orthogonal dimension. As Bloom's taxonomy is a tool widely used in biology education by researchers and instructors, it is important to examine the underlying assumptions embedded within how people may implicitly understand and use the taxonomy. In this paper, we empirically examine two major assumptions: the independence of the knowledge-type and cognitive-process dimensions and the use of action verbs as proxies for different cognitive processes. Contingency analysis on 940 assessment items revealed that the knowledge-type and cognitive-process dimensions are related and not independent. Subsequent correspondence analysis identified two principle axes in how the two dimensions are related, with three clusters of knowledge types and cognitive processes. Using the Shannon evenness index, we did not find a clear relationship between question prompt words (including action verbs) and cognitive processes in the assessment items. Based on these results, we suggest that both dimensions of the revised Bloom's taxonomy should be used and that question prompt words or action verbs alone are not sufficient in classifying the embedded learning objectives within assessment items.

Prevalence and Characterization of Quinolone Resistance in Campylobacter spp. Isolates in Chicken Livers from Retail Stores in Georgia, USA.

ABSTRACT: Campylobacter is a bacterial pathogen that causes human foodborne illnesses worldwide, and outbreaks have been associated with consumption of undercooked chicken livers. The objectives of this study were to compare two PCR assays of 250 Campylobacter isolates for identification to species, to assess antibiotic resistance of the isolates, and to analyze genetic diversity of the quinolone resistance determining regions (QRDRs) of the isolates. A double-blind design was used to identify the species of Campylobacter; 181 (72%) of the isolates were identified as Campylobacter jejuni, and 69 (28%) isolates were identified as Campylobacter coli by both PCR assays. A total of 93 (37.2%) isolates were resistant to at least one antibiotic. Among 88 C. jejuni isolates, 33 (18%) were resistant to nalidixic acid (NAL) and ciprofloxacin (CIP), 25 (14%) were resistant to tetracycline (TET), and 18 (10%) were resistant to NAL and TET. Two C. jejuni isolates were resistant to four of the tested antibiotics, and one isolate was resistant to five antibiotics. Two C. coli isolates were resistant to TET, and two were resistant to NAL, CIP, and TET. The amino acid sequences of the QRDRs for the isolates had eight point mutations and could be classified into 12 groups. Thirty-eight C. jejuni isolates resistant to NAL and CIP had a point mutation at residue 86 (substitution from threonine to isoleucine). However, six isolates without this substitution were resistant to NAL and/or CIP. Ten isolates with a point mutation at residue 86 were susceptible to NAL and CIP. This observation suggests that in addition to the substitution at residue 86 other mechanisms may confer resistance to quinolones. Further studies are needed to elucidate mechanisms for quinolone resistance in Campylobacter. The Campylobacter spp. isolated from chicken livers in this study were resistant to quinolones and other classes of antibiotics.