Remote monitoring of parental incubation conditions in the greater sandhill crane

To monitor incubation conditions in nests of greater sandhill cranes, a radiotrans-mitting egg was built using six temperature sensors, a position sensor, and a light sensor. Sensor readings were received, along with time of observations, and stored in a computer. The egg was used to monitor incubation in nests of six pairs of cranes during 1987 and 1988. Ambient temperature was also measured. Analysis of covariance (ANCOVA) was used to relate highest egg temperature, core egg temperature, and lowest egg temperature to ambient temperature, time since the egg was last turned, and time since the beginning of incubation. Ambient temperature had the greatest effect on egg temperature (P < 0.0001), followed by the time since the beginning of incubation and time since the egg was last turned. Pair effect, the class variable in the ANCOVA, was also very significant (P < 0.0001). A nine-term Fourier series was used to estimate the average core egg temperature versus time of day and was found to fit the data well (r² = 0.94). The Fourier series will be used to run a mechanical incubator to simulate natural incubation conditions for cranes. © 1995 Wiley-Liss, Inc.     

Landscape structure influences pollinator movements and directly affects plant reproductive success

Pollinators play a key role within most terrestrial communities in maintaining plant populations, as well as pollinating many agricultural crops for seed and fruit production. The mobility of pollinating animals is significant to their importance but we know little about how landscape structure influences pollinator movements. Linear landscape features such as hedgerows and embankments are conspicuous features of agricultural landscape structure and are important artificial habitats in their own right. However, there has been some debate as to the function of these landscape elements as corridors between larger expanses of semi‐natural habitat separated by urban and agricultural habitats. Few studies have specifically studied insect flight responses to linear landscape elements. By observing bumblebee flight behaviour along hedgerows and, by creating a medium‐scale experimental array of flower patches using an artificial linear feature, we examined whether such structures can elicit an oriented flight response along them and therefore facilitate insect movement through the landscape. We found that both hedgerows and artificial linear landscape features can influence the flight directions of bumblebees (Bombus spp. Hymenoptera, Apidae), one of Europe's most important groups of pollinators. A bioassay experiment in which Salvia pratensis (Lamiaceae) was planted into landscape patches with differing numbers of connecting hedgerows showed that this directional response can have a profound effect on plant reproductive success – plants had increased pollinator activity, pollen receipt and subsequent seed set in patches with more connections. The overall hedgerow connectedness of a landscape is therefore important both to bumblebee movement and to those plants which depend on bumblebees for pollination services.     

Detection of Mycobacterium tuberculosis Peptides in the Exosomes of Patients with Active and Latent M. tuberculosis Infection Using MRM-MS

The identification of easily measured, accurate diagnostic biomarkers for active tuberculosis (TB) will have a significant impact on global TB control efforts. Because of the host and pathogen complexities involved in TB pathogenesis, identifying a single biomarker that is adequately sensitive and specific continues to be a major hurdle. Our previous studies in models of TB demonstrated that exosomes, such as those released from infected macrophages, contain mycobacterial products, including many Mtb proteins. In this report, we describe the development of targeted proteomics assays employing multiplexed multiple reaction monitoring mass spectrometry (MRM-MS) in order to allow us to follow those proteins previously identified by western blot or shotgun mass spectrometry, and enhance biomarker discovery to include detection of Mtb proteins in human serum exosomes. Targeted MRM-MS assays were applied to exosomes isolated from human serum samples obtained from culture-confirmed active TB patients to detect 76 peptides representing 33 unique Mtb proteins. Our studies revealed the first identification of bacteria-derived biomarker candidates of active TB in exosomes from human serum. Twenty of the 33 proteins targeted for detection were found in the exosomes of TB patients, and included multiple peptides from 8 proteins (Antigen 85B, Antigen 85C, Apa, BfrB, GlcB, HspX, KatG, and Mpt64). Interestingly, all of these proteins are known mycobacterial adhesins and/or proteins that contribute to the intracellular survival of Mtb. These proteins will be included as target analytes in future validation studies as they may serve as markers for persistent active and latent Mtb infection. In summary, this work is the first step in identifying a unique and specific panel of Mtb peptide biomarkers encapsulated in exosomes and reveals complex biomarker patterns across a spectrum of TB disease states.     

In vivo tracking of transplanted mononuclear cells using manganese-enhanced magnetic resonance imaging (MEMRI)

Background

Transplantation of mononuclear cells (MNCs) has previously been tested as a method to induce therapeutic angiogenesis to treat limb ischemia in clinical trials. Non-invasive high resolution imaging is required to track the cells and evaluate clinical relevance after cell transplantation. The hypothesis that MRI can provide in vivo detection and long-term observation of MNCs labeled with manganese contrast-agent was investigated in ischemic rat legs.

Methods and Findings

The Mn-labeled MNCs were evaluated using 7-tesla high-field magnetic resonance imaging (MRI). Intramuscular transplanted Mn-labeled MNCs were visualized with MRI for at least 7 and up to 21 days after transplantation in the ischemic leg. The distribution of Mn-labeled MNCs was similar to that of ¹¹¹In-labeled MNCs measured with single-photon emission computed tomography (SPECT) and DiI-dyed MNCs with fluorescence microscopy. In addition, at 1–2 days after transplantation the volume of the site injected with intact Mn-labeled MNCs was significantly larger than that injected with dead MNCs, although the dead Mn-labeled MNCs were also found for approximately 2 weeks in the ischemic legs. The area covered by CD31-positive cells (as a marker of capillary endothelial cells) in the intact Mn-MNCs implanted site at 43 days was significantly larger than that at a site implanted with dead Mn-MNCs.

Conclusions

The present Mn-enhanced MRI method enabled visualization of the transplanted area with a 150–175 µm in-plane spatial resolution and allowed the migration of labeled-MNCs to be observed for long periods in the same subject. After further optimization, MRI-based Mn-enhanced cell-tracking could be a useful technique for evaluation of cell therapy both in research and clinical applications.     

Meta-analysis of genome-wide scans for total body BMD in children and adults reveals allelic heterogeneity and age-specific effects at the WNT16 locus

To identify genetic loci influencing bone accrual, we performed a genome-wide association scan for total-body bone mineral density (TB-BMD) variation in 2,660 children of different ethnicities. We discovered variants in 7q31.31 associated with BMD measurements, with the lowest P = 4.1 × 10(-11) observed for rs917727 with minor allele frequency of 0.37. We sought replication for all SNPs located ± 500 kb from rs917727 in 11,052 additional individuals from five independent studies including children and adults, together with de novo genotyping of rs3801387 (in perfect linkage disequilibrium (LD) with rs917727) in 1,014 mothers of children from the discovery cohort. The top signal mapping in the surroundings of WNT16 was replicated across studies with a meta-analysis P = 2.6 × 10(-31) and an effect size explaining between 0.6%-1.8% of TB-BMD variance. Conditional analyses on this signal revealed a secondary signal for total body BMD (P = 1.42 × 10(-10)) for rs4609139 and mapping to C7orf58. We also examined the genomic region for association with skull BMD to test if the associations were independent of skeletal loading. We identified two signals influencing skull BMD variation, including rs917727 (P = 1.9 × 10(-16)) and rs7801723 (P = 8.9 × 10(-28)), also mapping to C7orf58 (r(2) = 0.50 with rs4609139). Wnt16 knockout (KO) mice with reduced total body BMD and gene expression profiles in human bone biopsies support a role of C7orf58 and WNT16 on the BMD phenotypes observed at the human population level. In summary, we detected two independent signals influencing total body and skull BMD variation in children and adults, thus demonstrating the presence of allelic heterogeneity at the WNT16 locus. One of the skull BMD signals mapping to C7orf58 is mostly driven by children, suggesting temporal determination on peak bone mass acquisition. Our life-course approach postulates that these genetic effects influencing peak bone mass accrual may impact the risk of osteoporosis later in life.     

Modeling the temperature response of nitrification

To model nitrification rates in soils, it is necessary to have equations that accurately describe the effect of environmental variables on nitrification rates. A variety of equations have been used previously to describe the effect of temperature on rates of microbial processes. It is not clear which of these best describes the influence of temperature on nitrification rates in soil. I compared five equations for describing the effects of temperature on nitrification in two soils with very different temperature optima from a California oak woodland-annual grassland. The most appropriate equation depended on the range of temperatures being evaluated. A generalized Poisson density function best described temperature effects on nitrification rates in both soils over the range of 5 to 50 °C; however, the Arrhenius equation best described temperature effects over the narrower range of soil temperatures that normally occurs in the ecosystem (5 to 28 °C). Temperature optima for nitrification in most of the soils were greater than even the highest soil temperatures recorded at the sites. A model accounting for increased maintenance energy requirements at higher temperatures demonstrates how net energy production, rather than the gross energy production from nitrification, is maximized during adaptation by nitrifier populations to soil temperatures.     

Architectural design influences the diversity and structure of the built environment microbiome

Buildings are complex ecosystems that house trillions of microorganisms interacting with each other, with humans and with their environment. Understanding the ecological and evolutionary processes that determine the diversity and composition of the built environment microbiome—the community of microorganisms that live indoors—is important for understanding the relationship between building design, biodiversity and human health. In this study, we used high-throughput sequencing of the bacterial 16S rRNA gene to quantify relationships between building attributes and airborne bacterial communities at a health-care facility. We quantified airborne bacterial community structure and environmental conditions in patient rooms exposed to mechanical or window ventilation and in outdoor air. The phylogenetic diversity of airborne bacterial communities was lower indoors than outdoors, and mechanically ventilated rooms contained less diverse microbial communities than did window-ventilated rooms. Bacterial communities in indoor environments contained many taxa that are absent or rare outdoors, including taxa closely related to potential human pathogens. Building attributes, specifically the source of ventilation air, airflow rates, relative humidity and temperature, were correlated with the diversity and composition of indoor bacterial communities. The relative abundance of bacteria closely related to human pathogens was higher indoors than outdoors, and higher in rooms with lower airflow rates and lower relative humidity. The observed relationship between building design and airborne bacterial diversity suggests that we can manage indoor environments, altering through building design and operation the community of microbial species that potentially colonize the human microbiome during our time indoors.     

The human serum proteome: display of nearly 3700 chromatographically separated protein spots on two-dimensional electrophoresis gels and identification of 325 distinct proteins

Plasma, the soluble component of the human blood, is believed to harbor thousands of distinct proteins, which originate from a variety of cells and tissues through either active secretion or leakage from blood cells or tissues. The dynamic range of plasma protein concentrations comprises at least nine orders of magnitude. Proteins involved in coagulation, immune defense, small molecule transport, and protease inhibition, many of them present in high abundance in this body fluid, have been functionally characterized and associated with disease processes. For example, protein sequence mutations in coagulation factors cause various serious disease states. Diagnosing and monitoring such diseases in blood plasma of affected individuals has typically been conducted by use of enzyme-linked immunosorbent assays, which using a specific antibody quantitatively measure only the affected protein in the tested plasma samples. The discovery of protein biomarkers in plasma for diseases with no known correlations to genetic mutations is challenging. It requires a highly parallel display and quantitation strategy for proteins. We fractionated blood serum proteins prior to display on two-dimensional electrophoresis (2-DE) gels using immunoaffinity chromatography to remove the most abundant serum proteins, followed by sequential anion-exchange and size-exclusion chromatography. Serum proteins from 74 fractions were displayed on 2-DE gels. This approach succeeded in resolving approximately 3700 distinct protein spots, many of them post-translationally modified variants of plasma proteins. About 1800 distinct serum protein spots were identified by mass spectrometry. They collapsed into 325 distinct proteins, after sequence homology and similarity searches were carried out to eliminate redundant protein annotations. Although a relatively insensitive dye, Coomassie Brilliant Blue G-250, was used to visualize protein spots, several proteins known to be present in serum in < 10 ng/mL concentrations were identified such as interleukin-6, cathepsins, and peptide hormones. Considering that our strategy allows highly parallel protein quantitation on 2-DE gels, it holds promise to accelerate the discovery of novel serum protein biomarkers.     

EFFECTS OF METAL AND PETROLEUM HYDROCARBON CONTAMINATION ON BENTHIC DIATOM COMMUNITIES NEAR CASEY STATION,ANTARCTICA: AN EXPERIMENTAL APPROACH1

The effects of metals and petroleum hydrocarbons on benthic marine diatom communities were assessed using a manipulative field experiment at Casey Station, Antarctica. Uncontaminated, metal contaminated, and petroleum hydrocarbon contaminated sediments were deployed for 11 weeks during the 1999 austral summer. The treatments were deployed at three different locations: Brown Bay, which has elevated levels of anthropogenic contaminants, and two uncontaminated reference locations, O'Brien Bay and Sparkes Bay, the latter of which has naturally occurring high concentrations of some heavy metals. At each location, significant differences between the composition of diatom communities recruiting to control and petroleum hydrocarbon contaminated treatments were observed. Navicula directa (Smith) Ralfs occurred at lower abundances in the petroleum hydrocarbon contaminated treatments than in the control treatments. In contrast, Navicula cancellata Donkin occurred at higher relative abundances in both contaminated treatments relative to the control treatment. Interactions between treatment and location were also observed for several species, including Navicula glaciei Van Heurck. Significant differences in the overall community composition of diatom communities between control and metal contaminated treatments and metal contaminated and petroleum hydrocarbon contaminated treatments were only observed within Brown Bay. The location of deployment also had a significant influence on the composition of diatom communities. Brown Bay had higher abundances of Achnanthes brevipes Agardh but lower abundances of Navicula aff. cincta (Ehrenberg) Van Heurck than either reference locations. This experiment demonstrated that benthic diatom communities are sensitive to sediment contamination and would be suitable for future monitoring work within this and other areas of Antarctica.