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Khristian Alegría 

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February 07, 2023
Campus News

In Memoriam: Former Carnegie research tech dies at 75

Youying Dong

Youying Dong

Graduate Student

Abstract
The intestines of animals are colonized by commensal microbes, which impact host development, health, and behavior. Precise quantification of colonization is essential for studying the complex interactions between host and microbe both to validate the microbial composition and study its effects. Drosophila melanogaster,which has a low native microbial diversity and is economical to rear with defined microbial composition, has emerged as a model organism for studying the gut microbiome. Analyzing the microbiome of an individual organism requires identification of which microbial species are present and quantification of their absolute abundance. This article presents a method for the analysis of a large number of individual fly microbiomes. The flies are prepared in 96-well plates, enabling the handling of a large number of samples at once. Microbial abundance is quantified by plating up to 96 whole fly homogenates on a single agar plate in an array of spots and then counting the colony forming units (CFUs) that grow in each spot. This plating system is paired with an automated CFU quantification platform, which incorporates photography of the plates, differentiation of fluorescent colonies, and automated counting of the colonies using an ImageJ plugin. Advantages are that (i) this method is sensitive enough to detect differences between treatments, (ii) the spot plating method is as accurate as traditional plating methods, and (iii) the automated counting process is accurate and faster than manual counting. The workflow presented here enables high-throughput quantification of CFUs in a large number of replicates and can be applied to other microbiology study systems including in vitro and other small animal models.
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Abstract
Windthrows (trees uprooted and broken by winds) are common across the Amazon. They range in size from single trees to large gaps that lead to changes in forest dynamics, composition, structure, and carbon balance. Yet, the current understanding of the spatial variability of windthrows is limited. By integrating remote sensing data and geospatial analysis, we present the first study to examine the occurrence, area, and direction of windthrows and the control that environmental variables exert on them across the whole Amazon. Windthrows are more frequent and larger in the northwestern Amazon (Peru and Colombia), with the central Amazon (Brazil) being another hot spot of windthrows. The predominant direction of windthrows is westward. Rainfall, surface elevation, and soil characteristics explain the variability (20%-50%) of windthrows but their effects vary regionally. A better understanding of the spatial dynamics of windthrows will improve understanding of the functioning of Amazon forests.
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Abstract
Crystal structures of minerals are defined by a specific atomic arrangement within the unit-cell, which follows the laws of symmetry specific to each crystal system. The causes for a mineral to crystallize in a given crystal system have been the subject of many studies showing their dependency on different formation conditions, such as the presence of aqueous fluids, biotic activity and many others. Different attempts have been made to quantify and interpret the information that we can gather from studying crystal symmetry and its distribution in the mineral kingdom. However, these methods are mostly outdated or at least not compatible for use on large datasets available today. Therefore, a revision of symmetry index calculation has been made in accordance with the growing understanding of mineral species and their characteristics. In the gathered data, we observe a gradual but significant decrease in crystal symmetry through the stages of mineral evolution, from the formation of the solar system to modern day. However, this decrease is neither uniform nor linear, which provides further implications for mineral evolution from the viewpoint of crystal symmetry. The temporal distribution of minerals based on the number of essential elements in their chemical formulae and their symmetry index has been calculated and compared to explore their behaviour. Minerals with four to eight essential elements have the lowest average symmetry index, while being the most abundant throughout all stages of mineral evolution. There are many open questions, including those pertaining to whether or not biological activity on Earth has influenced the observed decrease in mineral symmetry through time and whether or not the trajectory of planetary evolution of a geologically active body is one of decreasing mineral symmetry/increasing complexity.
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Abstract
Nitrification controls the oxidation state of bioavailable nitrogen. Distinct clades of chemoautotrophic microorganisms - predominantly ammonia-oxidizing archaea (AOA) and nitrite-oxidizing bacteria (NOB) - regulate the two steps of nitrification in the ocean, but explanations for their observed relative abundances and nitrification rates remain incomplete and their contributions to the global marine carbon cycle via carbon fixation remain unresolved. Using a mechanistic microbial ecosystem model with nitrifying functional types, we derive simple expressions for the controls on AOA and NOB in the deep, oxygenated open ocean. The relative biomass yields, loss rates, and cell quotas of AOA and NOB control their relative abundances, though we do not need to invoke a difference in loss rates to explain the observed relative abundances. The supply of ammonium, not the traits of AOA or NOB, controls the relatively equal ammonia and nitrite oxidation rates at steady state. The relative yields of AOA and NOB alone set their relative bulk carbon fixation rates in the water column. The quantitative relationships are consistent with multiple in situ datasets. In a complex global ecosystem model, nitrification emerges dynamically across diverse ocean environments, and ammonia and nitrite oxidation and their associated carbon fixation rates are decoupled due to physical transport and complex ecological interactions in some environments. Nevertheless, the simple expressions capture global patterns to first order. The model provides a mechanistic upper estimate on global chemoautotrophic carbon fixation of 0.2-0.5 Pg C yr(-1), which is on the low end of the wide range of previous estimates. Modeled carbon fixation by AOA (0.2-0.3 Pg C yr(-1)) exceeds that of NOB (about 0.1 Pg C yr(-1)) because of the higher biomass yield of AOA. The simple expressions derived here can be used to quantify the biogeochemical impacts of additional metabolic pathways (i.e., mixotrophy) of nitrifying clades and to identify alternative metabolisms fueling carbon fixation in the deep ocean.
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Abstract
Synchrotron X-ray powder diffraction and infrared (IR) spectroscopy studies on natural brucite were conducted up to 31 GPa using diamond-anvil cell (DAC) techniques at beamlines X17C and U2A of National Synchrotron Light Source (NSLS). The lattice parameters and unit-cell volumes were refined in P (3) over bar m1 space group throughout the experimental pressure range. The anisotropy of lattice compression decreases with pressure due to a more compressible c axis and the compression becomes nearly isotropic in the pressure range of 10-25 GPa. The unit-cell volumes are fitted to the third-order Birch-Mumaghan equation of state, yielding K-0 = 39.4(1.3) GPa, K-0' = 8.4(0.4) for the bulk modulus and its pressure-derivative, respectively. No phase transition or amorphization was resolved from the X-ray diffraction data up to 29 GPa, however, starting from 4 GPa, a new infrared vibration band (similar to 3638 cm(-1)) 60 cm(-1) below the OH stretching A(2u) band of brucite was found to coexist with the A(2u) band and its intensity continuously increases with pressure. The new OH stretching band has a more pronounced redshift as a function of pressure (-4.7 cm(-1)/GPa) than the A(2u) band (-0.7 cm(-1)/GPa). Comparison with first-principles calculations suggests that a structural change involving the disordered H sublattice is capable of reconciling the observations from X-ray diffraction and infrared spectroscopy studies.
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Abstract
Zebrafish are a valuable model for mammalian lipid metabolism; larvae process lipids similarly through the intestine and hepatobiliary system and respond to drugs that block cholesterol synthesis in humans. After ingestion of fluorescently quenched phospholipids, endogenous lipase activity and rapid transport of cleavage products results in intense gall bladder fluorescence. Genetic screening identifies zebrafish mutants, such as fat free, that show normal digestive organ morphology but severely reduced phospholipid and cholesterol processing. Thus, fluorescent lipids provide a sensitive readout of lipid metabolism and are a powerful tool for identifying genes that mediate vertebrate digestive physiology.
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