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1.
Biotechniques ; 75(4): 150-156, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37671637

RESUMO

Recombinant rabbit monoclonal antibodies (rabbit rAbs) have shown promise in various biomedical fields. However, it is challenging and costly to generate rabbit rAbs using traditional techniques. Here we describe a convenient and cost-effective method. Using this method, we generated rabbit rAbs against mouse soluble IL-6 receptor α with affinities in the range of 10-9 to 10-12 M. The presented method is suitable for industrial and academic scientists looking to customize rabbit rAbs for their research.

2.
Chemosphere ; 334: 138872, 2023 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-37182716

RESUMO

In terrestrial ecosystems, the nitrogen dynamics, including N2O production, are majorly regulated by a complex consortium of microbes favored by different substrates and environmental conditions. To better predict the daily, seasonal and annual variation in N2O fluxes, it is critical to estimate the temperature sensitivity of different microbial groups for N2O fluxes under oxic and suboxic conditions prevalent in soil and wetlands. Here, we studied the temperature sensitivity of two groups of ammonia oxidizers, archaea (AOA) and bacteria (AOB), in relation to N2O fluxes through both nitrification and nitrifier-denitrification pathways across a wide temperature gradient (10-55 °C). Using square root theory (SQRT) and macromolecular rate theory (MMRT) models, we estimated thermodynamic parameters and cardinal temperatures, including maximum temperature sensitivity (TSmax). The distinction between N2O pathways was facilitated by microbial-specific inhibitors (PTIO and C2H2) and controlled oxygen supply environments (oxic: ambient level; and suboxic: ∼4%). We found that nitrification supported by AOA (NtA) and AOB (NtB) dominated N2O production in an oxic climate, while only AOB-supported nitrifier-denitrification (NDB) majorly contributed (>90%) to suboxic N2O budget. The models predicted significantly higher optimum temperature (Topt) and TSmax for NtA and NDB compared to NtB. Intriguingly, both NtB and NDB exhibited significantly wider temperature ranges than NtA. Altogether, our results suggest that temperature and oxygen supply control the dominance of specific AOA- and AOB-supported N2O pathways in soil and sediments. This emergent understanding can potentially contribute toward novel targeted N2O inhibitors for GHG mitigation under global warming.


Assuntos
Amônia , Ecossistema , Amônia/metabolismo , Oxirredução , Microbiologia do Solo , Archaea/metabolismo , Nitrificação , Solo/química , Temperatura , Oxigênio/metabolismo , Óxido Nitroso/análise
3.
Sci Total Environ ; 882: 163412, 2023 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-37059149

RESUMO

Microbes are a critical component of soil ecosystems, performing crucial functions in biogeochemical cycling, carbon sequestration, and plant health. However, it remains uncertain how their community structure, functioning, and resultant nutrient cycling, including net GHG fluxes, would respond to climate change at different scales. Here, we review global and regional climate change effects on soil microbial community structure and functioning, as well as the climate-microbe feedback and plant-microbe interactions. We also synthesize recent studies on climate change impacts on terrestrial nutrient cycles and GHG fluxes across different climate-sensitive ecosystems. It is generally assumed that climate change factors (e.g., elevated CO2 and temperature) will have varying impacts on the microbial community structure (e.g., fungi-to-bacteria ratio) and their contribution toward nutrient turnover, with potential interactions that may either enhance or mitigate each other's effects. Such climate change responses, however, are difficult to generalize, even within an ecosystem, since they are subjected to not only a strong regional influence of current ambient environmental and edaphic conditions, historical exposure to fluctuations, and time horizon but also to methodological choices (e.g., network construction). Finally, the potential of chemical intrusions and emerging tools, such as genetically engineered plants and microbes, as mitigation strategies against global change impacts, particularly for agroecosystems, is presented. In a rapidly evolving field, this review identifies the knowledge gaps complicating assessments and predictions of microbial climate responses and hindering the development of effective mitigation strategies.


Assuntos
Mudança Climática , Microbiologia do Solo , Dióxido de Carbono/análise , Temperatura Alta , Erosão do Solo , Bioengenharia , Bactérias/genética
4.
Nanoscale Res Lett ; 17(1): 104, 2022 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-36315294

RESUMO

Highly sensitive, simple and reliable colorimetric probe for Cu2+-ion detection was visualized with the L-cysteine functionalized gold nanoparticle (LS-AuNP) probes. The pronounced sensing of Cu2+ with high selectivity was rapidly featured with obvious colour change that enabled to visually sense Cu2+ ions by naked eyes. By employing systemic investigations on crystallinities, elemental compositions, microstructures, surface features, light absorbance, zeta potentials and chemical states of LS-AuNP probes, the oxidation-triggered aggregation effect of LS-AuNP probes was envisioned. The results indicated that the mediation of Cu2+ oxidation coordinately caused the formation of disulfide cystine, rendering the removal of thiol group at AuNPs surfaces. These features reflected the visual colour change for the employment of tracing Cu2+ ions in a quantitative way.

5.
Sci Rep ; 12(1): 16456, 2022 09 30.
Artigo em Inglês | MEDLINE | ID: mdl-36180528

RESUMO

Growing evidence suggests that the origins of the panzootic amphibian pathogens Batrachochytrium dendrobatidis (Bd) and Batrachochytrium salamandrivorans (Bsal) are in Asia. In Taiwan, an island hotspot of high amphibian diversity, no amphibian mass mortality events linked to Bd or Bsal have been reported. We conducted a multi-year study across this subtropical island, sampling 2517 individuals from 30 species at 34 field sites, between 2010 and 2017, and including 171 museum samples collected between 1981 and 2009. We analyzed the skin microbiome of 153 samples (6 species) from 2017 in order to assess any association between the amphibian skin microbiome and the probability of infection amongst different host species. We did not detect Bsal in our samples, but found widespread infection by Bd across central and northern Taiwan, both taxonomically and spatially. Museum samples show that Bd has been present in Taiwan since at least 1990. Host species, geography (elevation), climatic conditions and microbial richness were all associated with the prevalence of infection. Host life-history traits, skin microbiome composition and phylogeny were associated with lower prevalence of infection for high altitude species. Overall, we observed low prevalence and burden of infection in host populations, suggesting that Bd is enzootic in Taiwan where it causes subclinical infections. While amphibian species in Taiwan are currently threatened by habitat loss, our study indicates that Bd is in an endemic equilibrium with the populations and species we investigated. However, ongoing surveillance of the infection is warranted, as changing environmental conditions may disturb the currently stable equilibrium.


Assuntos
Quitridiomicetos , Microbiota , Micoses , Anfíbios , Animais , Batrachochytrium , Humanos , Micoses/epidemiologia , Taiwan/epidemiologia
6.
Sci Total Environ ; 853: 158611, 2022 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-36087665

RESUMO

Mountains are an essential component of the global life-support system. They are characterized by a rugged, heterogenous landscape with rapidly changing environmental conditions providing myriad ecological niches over relatively small spatial scales. Although montane species are well adapted to life at extremes, they are highly vulnerable to human derived ecosystem threats. Here we build on the manifesto 'World Scientists' Warning to Humanity', issued by the Alliance of World Scientists, to outline the major threats to mountain ecosystems. We highlight climate change as the greatest threat to mountain ecosystems, which are more impacted than their lowland counterparts. We further discuss the cascade of "knock-on" effects of climate change such as increased UV radiation, altered hydrological cycles, and altered pollution profiles; highlighting the biological and socio-economic consequences. Finally, we present how intensified use of mountains leads to overexploitation and abstraction of water, driving changes in carbon stock, reducing biodiversity, and impacting ecosystem functioning. These perturbations can provide opportunities for invasive species, parasites and pathogens to colonize these fragile habitats, driving further changes and losses of micro- and macro-biodiversity, as well further impacting ecosystem services. Ultimately, imbalances in the normal functioning of mountain ecosystems will lead to changes in vital biological, biochemical, and chemical processes, critically reducing ecosystem health with widespread repercussions for animal and human wellbeing. Developing tools in species/habitat conservation and future restoration is therefore essential if we are to effectively mitigate against the declining health of mountains.


Assuntos
Biodiversidade , Ecossistema , Animais , Humanos , Mudança Climática , Água , Carbono , Conservação dos Recursos Naturais
7.
Chemosphere ; 289: 133049, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34838835

RESUMO

Understanding the environmental niche segregation of ammonia-oxidizing archaea (AOA) and bacteria (AOB) and its impact on their relative contributions to nitrification and nitrous oxide (N2O) production is essential for predicting N2O dynamics within an ecosystem. Here, we used ammonia oxidizer-specific inhibitors to measure the differential contributions of AOA and AOB to potential ammonia oxidization (PAO) and N2O fluxes over pH (4.0-9.0) and temperature (10-45 °C) gradients in five soils and three wetland sediments. AOA and AOB activities were differentiated using PTIO (2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide), 1-octyne, and acetylene. We used square root growth (SQRT) and macromolecular rate theory (MMRT) models to estimate cardinal temperatures and thermodynamic characteristics for AOA- and AOB-dominated PAO and N2O fluxes. We found that AOA and AOB occupied different niches for PAO, and soil temperature was the major determinant of niche specialization. SQRT and MMRT models predicted a higher optimum temperature for AOA-dominated PAO and N2O fluxes compared with those of AOB. Additionally, PAO was dominated by AOA in acidic conditions, whereas both AOA- and AOB-dominated N2O fluxes decreased with increasing pH. Consequently, net N2O fluxes (AOA and AOB) under acidic conditions were approximately one to three-fold higher than those observed in alkaline conditions. Moreover, structural equation and linear regression modeling confirmed a significant positive correlation (R2 = 0.45, p < 0.01) between PAO and N2O fluxes. Collectively, these results show the influence of ammonia oxidizer responses to temperature and pH on nitrification-driven N2O fluxes, highlighting the potential for mitigating N2O emissions via pH manipulation.


Assuntos
Amônia , Solo , Archaea , Ecossistema , Nitrificação , Oxirredução , Microbiologia do Solo
8.
Artigo em Inglês | MEDLINE | ID: mdl-34281066

RESUMO

Nitrification inhibitors (NIs) such as dicyandiamide (DCD), 3,4-dimethylpyrazole phosphate (DMPP), and allylthiourea (AT) are commonly used to suppress ammonia oxidization at different time scales varying from a few hours to several months. Although the responses of NIs to edaphic and temperature conditions have been studied, the influence of the aforementioned factors on their inhibitory effect remains unknown. In this study, laboratory-scale experiments were conducted to assess the short-term (24 h) influence of eight abiotic and biotic factors on the inhibitory effects of DCD, DMPP, and AT across six cropped and non-cropped soils at two temperature conditions with three covariates of soil texture. Simultaneously, the dominant contributions of ammonia-oxidizing archaea (AOA) and bacteria (AOB) to potential ammonia oxidization (PAO) were distinguished using the specific inhibitor 2 phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide (PTIO). Our results revealed that AT demonstrated a considerably greater inhibitory effect (up to 94.9% for an application rate of 75 mg of NI/kg of dry soil) than DCD and DMPP. The inhibitory effect of AT was considerably affected by the relative proportions of silt, sand, and clay in the soil and total PAO. In contrast to previous studies, the inhibitory effects of all three NIs remained largely unaffected by the landcover type and temperature conditions for the incubation period of 24 h. Furthermore, the efficacy of all three tested NIs was not affected by the differential contributions of AOA and AOB to PAO. Collectively, our results suggested a limited influence of temperature on the inhibitory effects of all three NIs but a moderate dependence of AT on the soil texture and PAO. Our findings can enhance the estimation of the inhibitory effect in soil, and pure cultures targeting the AOA and AOB supported ammonia oxidization and, hence, nitrogen dynamics under NI applications.


Assuntos
Nitrificação , Solo , Amônia/análise , Guanidinas , Oxirredução , Fosfatos , Pirazóis , Microbiologia do Solo , Tioureia/análogos & derivados
9.
Ecotoxicol Environ Saf ; 220: 112383, 2021 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-34082242

RESUMO

Geochemical approaches are popular for evaluations based on heavy metal concentrations in sediments or soils for eco-risk assessment. This study proposes a systematic geochemical approach (SymGeo) to explore six heavy metals in topsoils and bird tissues and organs of the target birds. We assume that the proposed approach based on field-collected heavy metals in topsoils and feathers can predict the areas with the potential risk of the heavy metals in birds. Finite mixture distribution modeling (FMDM) was used to identify background values of the heavy metal concentrations in topsoil. A spatial enrichment factor (EF), potential contamination index (PCI), contamination degree (Cod), and potential ecological risk index (PRI) based on FMDM results for topsoil, and a potential risk index (PRIbird) of heavy metals in the birds, were utilized for systematic prioritization of high eco-risk areas. Using multiple EF, PRI, and Cod results and multiple PRI-based maps of the heavy metals in feathers, we systematically prioritized risk areas where there is a high potential for heavy metal contamination in the birds. Our results indicate that heavy metal concentrations in the feather, liver, and kidney are not spatially cross-autocorrelated but are statistically significantly correlated with some heavy metals in topsoil due to external and internal depositions. Further, multiple EF, Cod, and RI distributions for topsoil, along with the PRI of the feather, showed that adequate coverages for potential risk for birds were greater than 71.05% in the top 30% and 84.69% in the top 20% potential eco-risk priority area of heavy metals in bird liver and kidney. Hence, our proposed approach suggests that assessments of heavy metals in bird feathers and topsoils without bird organs can be utilized to identify spatially high-risk areas. The proposed approach could be improved by incorporating water and sediment samples to enhance the crowdsourcing and the species-specific data.


Assuntos
Aves , Monitoramento Ambiental/métodos , Plumas/química , Poluentes do Solo/química , Solo/química , Animais , China , Metais Pesados/análise , Medição de Risco
10.
J Environ Manage ; 294: 112886, 2021 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-34130136

RESUMO

Accurate information provided by reliable models is essential for identifying hotspots and mitigating roadkill. However, existing methods, such as kernel density estimation (KDE) and maximum entropy modeling (ME) may individually identify only a subset of the suitable locations for mitigation, because KDE cannot detect hotspots once local abundances are depressed, and ME may only partially identify current hotspots due to imperfect discrimination skill. Here, we propose a hybrid consensus modeling (HCM) approach that leverages the strengths of both KDE and ME by using their consensus to identify the core subset of hotspots. We collected herpetofauna (amphibians and reptiles) roadkill data (N = 839) along four roads in Taiwan (R.O.C.) to evaluate the statistical performance and theoretical mitigation efficiency of HCM, KDE and ME, and to compare the allocation among roads, spatial clustering, and environmental conditions in the identified hotspots. HCM was applied on the herpetofauna dataset as well as separately on amphibians and reptiles. Although the discrimination skill of KDE and ME models for both target clades together was good to excellent (AUCKDE = 0.944, AUCME = 0.822), the highest theoretical mitigation efficiency, was displayed by HCM Consensus (2.89), followed by KDE (2.58), and ME (1.91). Furthermore, we show that theoretical mitigation efficiency increases with decreasing spatial clustering (Moran's I). Given pervasive budget constraints, we recommend to limit permanent mitigation measures such as fenced culverts to HCM Consensus hotspots, temporary measures to KDE hotspots, and to target additional monitoring at ME hotspots.


Assuntos
Anfíbios , Répteis , Animais , Consenso , Taiwan
11.
Sci Total Environ ; 764: 144349, 2021 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-33412402

RESUMO

Soil encompasses diverse microbial communities that are essential for fundamental ecosystem functions such as biogeochemical cycling. To better understand underlying biogeochemical processes, it is necessary to know the structure of soil archaeal and bacterial communities and their responses to edaphic and climate variables within and across various land cover types (LCTs) and environmental zones (ENZs). Here we sampled eighty-nine sites across five ENZs and four LCTs within the Western Pacific region. Through leveraging the second-generation sequencing of topsoil samples, we showed that α-diversity (taxonomic diversity) of archaea strongly varied within LCTs, whereas bacterial α-diversity was significantly controlled by both LCT and ENZ. Soil archaea and bacteria showed global niche differentiation associated with contrasting diversity responses to latitude and differential responses of microbial diversity patterns to edaphic and climate variables within LCTs and ENZs. In contrast to α-diversity, microbial ß-diversity (the compositional dissimilarity between sites) was majorly governed by ENZs, particularly for archaea (P < 0.01). Our results highlight the importance of LCTs and ENZs for understanding soil microbial contributions to nutrient dynamics and ecosystem resilience under land-use intensification and climate change.


Assuntos
Ecossistema , Microbiota , Archaea/genética , Bactérias , Biodiversidade , Solo , Microbiologia do Solo
12.
Sci Total Environ ; 751: 141721, 2021 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-32861948

RESUMO

Globally, soils are subject to radical changes in their biogeochemistry as rampant deforestation and other forms of land use and climate change continue to transform planet Earth. To better understand soil ecosystem functioning, it is necessary to understand the responses of soil microbial diversity and community structure to changing climate, land cover, and associated environmental variables. With next-generation sequencing, we investigated changes in topsoil fungi community structure among different land cover types (from Forest to Cropland) and climate zones (from Hot to Cold zones) in the Western Pacific Region. We demonstrated that climate zones substantially (P = 0.001) altered the soil fungal beta-diversity (change in community composition), but not alpha-diversity (taxonomical diversity). In particular, precipitation, temperature, and also latitude were the best predictors of beta-diversity. Individual fungal classes displayed divergent but strong responses to climate variables and latitude, suggesting niche differentiation at lower taxonomic levels. We also demonstrated that fungal taxonomic diversity differentially responded to latitude across land covers: fungal diversity increased towards lower latitudes in the Forest and Cropland (R2 = 0.19) but increased towards both lower and higher latitudes in Fallow land (R2 = 0.45). Further, alpha-diversity was significantly influenced by soil pH in Forest (P = 0.02), and by diurnal temperature range in Fallow land and mean annual precipitation in Cropland. Collectively, various land cover types had differential influence on the latitude diversity gradient, while climate, and to some extent, edaphic variables, were crucial in shaping soil fungal community structure. Our results can also serve as a baseline for estimating global change impacts on fungal community structure in the Western Pacific Region.


Assuntos
Micobioma , Mudança Climática , Ecossistema , Florestas , Solo , Microbiologia do Solo
13.
PLoS One ; 15(12): e0243135, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33270722

RESUMO

Efficient biodiversity conservation requires that limited resources be allocated in accordance with national responsibilities and priorities. Without appropriate computational tools, the process of determining these national responsibilities and conservation priorities is time intensive when considering many species across geographic scales. Here, we have developed a computational tool as a module for the ArcGIS geographic information system. The ArcGIS National Responsibility Assessment Tool (NRA-Tool) can be used to create hierarchical lists of national responsibilities and priorities for global species conservation. Our tool will allow conservationists to prioritize conservation efforts and to focus limited resources on relevant species and regions. We showcase our tool with data on 258 bird species and various biophysical regions, including Environmental Zones in 58 Asian countries and regions. Our tool provides a decision support system for conservation policy with attractive and easily interpretable visual outputs illustrating national responsibilities and priorities for species conservation. The graphical output allows for smooth integration into assessment reports, such as the European Article 17 report, the Living Planet Index report, or similar regional and global reports.


Assuntos
Biodiversidade , Conservação dos Recursos Naturais , Sistemas de Informação Geográfica , Animais , Ásia , Monitorização de Parâmetros Ecológicos , Ecossistema , Europa (Continente)
14.
Sensors (Basel) ; 20(13)2020 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-32605303

RESUMO

Real-time identification of irrigation water pollution sources and pathways (PSP) is crucial to ensure both environmental and food safety. This study uses an integrated framework based on the Internet of Things (IoT) and the blockchain technology that incorporates a directed acyclic graph (DAG)-configured wireless sensor network (WSN), and GIS tools for real-time water pollution source tracing. Water quality sensors were installed at monitoring stations in irrigation channel systems within the study area. Irrigation water quality data were delivered to databases via the WSN and IoT technologies. Blockchain and GIS tools were used to trace pollution at mapped irrigation units and to spatially identify upstream polluted units at irrigation intakes. A Water Quality Analysis Simulation Program (WASP) model was then used to simulate water quality by using backward propagation and identify potential pollution sources. We applied a "backward pollution source tracing" (BPST) process to successfully and rapidly identify electrical conductivity (EC) and copper (Cu2+) polluted sources and pathways in upstream irrigation water. With the BPST process, the WASP model effectively simulated EC and Cu2+ concentration data to identify likely EC and Cu2+ pollution sources. The study framework is the first application of blockchain technology for effective real-time water quality monitoring and rapid multiple PSPs identification. The pollution event data associated with the PSP are immutable.

15.
Chemosphere ; 252: 126583, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32443269

RESUMO

Birds are bioindicators for research on the relationship between environmental heavy metal concentration levels and accumulation levels in bird tissues. We use roadkill samples, collected by citizen science participants, to investigate the accumulation levels and associations of seven heavy metals in internal organs (heart, liver, and kidney), feathers (primary and breast), and bones (sternum and femur) of two focal species, Amaurornis phoenicurus and Gallinula chloropus. We found that heavy metal accumulation varied by target tissue, and that variables are associated with bird species and heavy metal type. Although Zn and Cu were highest by concentration among both species, Cu was mostly accumulated in internal organs, As in feathers, and Pb in bones. Concentrations of As, Ni, and Pb in feathers of both focal species were lower than those reported in literature, whereas Cd and Cr were above toxic levels. The results also showed that spatial correlation for heavy metal concentration among bird tissues were weaker than non-spatial correlation, suggesting low spatial autocorrelations and variability. In addition, multiple regression analysis revealed significant correlation for Cr, As, and Pb estimations in A. phoenicurus heart, sternum, and kidney, respectively; and potentially Cr in G. chloropus femur by using feathers. These results support the feasibility of using feathers as indicators of As, Cr, and Pb heavy metal contamination to enhance our understanding of heavy metal accumulation in birds, although caution is required for feather-based estimations of Cd, Cu, and Ni concentration.


Assuntos
Aves/metabolismo , Monitoramento Ambiental/métodos , Metais Pesados/metabolismo , Animais , Crowdsourcing , Biomarcadores Ambientais , Plumas/química , Rim/química , Fígado/química , Metais Pesados/análise
16.
Biochim Biophys Acta Biomembr ; 1862(6): 183235, 2020 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-32126232

RESUMO

The adenosine 2A receptor (A2AR), a G-protein-coupled receptor (GPCR), was solubilised and purified encapsulated in styrene maleic acid lipid particles (SMALPs). The purified A2AR-SMALP was associated with phospholipids characteristic of the plasma membrane of Pichia pastoris, the host used for its expression, confirming that the A2AR-SMALP encapsulated native lipids. The fluorescence spectrum of the A2AR-SMALP showed a characteristic broad emission peak at 330 nm, produced by endogenous Trp residues. The inverse agonist ZM241385 caused 30% increase in fluorescence emission, unusually accompanied by a red-shift in the emission wavelength. The emission spectrum also showed sub-peaks at 321 nm, 335 nm and 350 nm, indicating that individual Trp inhabited different environments following ZM241385 addition. There was no effect of the agonist NECA on the A2AR-SMALP fluorescence spectrum. Substitution of two Trp residues by Tyr suggested that ZM241385 affected the environment and mobility of Trp2466.48 in TM6 and Trp2687.33 at the extracellular face of TM7, causing transition to a more hydrophobic environment. The fluorescent moiety IAEDANS was site-specifically introduced at the intracellular end of TM6 (residue 2316.33) to report on the dynamic cytoplasmic face of the A2AR. The inverse agonist ZM241385 caused a concentration-dependent increase in fluorescence emission as the IAEDANS moved to a more hydrophobic environment, consistent with closing the G-protein binding crevice. NECA generated only 30% of the effect of ZM241385. This study provides insight into the SMALP environment; encapsulation supported constitutive activity of the A2AR and ZM241385-induced conformational transitions but the agonist NECA generated only small effects.


Assuntos
Lipídeos/química , Receptor A2A de Adenosina/química , Estireno/química , Interações Hidrofóbicas e Hidrofílicas , Ligantes , Maleatos/química , Pichia/química , Conformação Proteica , Espectrometria de Fluorescência/métodos , Triazinas/farmacologia , Triazóis/farmacologia , Triptofano/química
17.
Sci Rep ; 9(1): 18712, 2019 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-31822696

RESUMO

The E. coli membrane protein ZipA, binds to the tubulin homologue FtsZ, in the early stage of cell division. We isolated ZipA in a Styrene Maleic Acid lipid particle (SMALP) preserving its position and integrity with native E. coli membrane lipids. Direct binding of ZipA to FtsZ is demonstrated, including FtsZ fibre bundles decorated with ZipA. Using Cryo-Electron Microscopy, small-angle X-ray and neutron scattering, we determine the encapsulated-ZipA structure in isolation, and in complex with FtsZ to a resolution of 1.6 nm. Three regions can be identified from the structure which correspond to, SMALP encapsulated membrane and ZipA transmembrane helix, a separate short compact tether, and ZipA globular head which binds FtsZ. The complex extends 12 nm from the membrane in a compact structure, supported by mesoscale modelling techniques, measuring the movement and stiffness of the regions within ZipA provides molecular scale analysis and visualisation of the early divisome.


Assuntos
Proteínas de Bactérias/metabolismo , Proteínas de Transporte/metabolismo , Proteínas de Ciclo Celular/metabolismo , Divisão Celular/fisiologia , Proteínas do Citoesqueleto/metabolismo , Proteínas de Escherichia coli/metabolismo , Proteínas de Bactérias/fisiologia , Proteínas de Transporte/fisiologia , Proteínas de Transporte/ultraestrutura , Proteínas de Ciclo Celular/fisiologia , Proteínas de Ciclo Celular/ultraestrutura , Microscopia Crioeletrônica/métodos , Proteínas do Citoesqueleto/fisiologia , Escherichia coli/metabolismo , Proteínas de Escherichia coli/fisiologia , Proteínas de Escherichia coli/ultraestrutura , Proteínas de Membrana/metabolismo , Ligação Proteica
18.
Microorganisms ; 7(11)2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31690001

RESUMO

Ammonia oxidizing archaea (AOA) and bacteria (AOB) are thought to contribute differently to soil nitrification, yet the extent to which their relative abundances influence the temperature response of nitrification is poorly understood. Here, we investigated the impact of different AOA to AOB ratios on soil nitrification potential (NP) across a temperature gradient from 4 °C to 40 °C in twenty different organic and inorganic fertilized soils. The temperature responses of different relative abundance of ammonia oxidizers for nitrification were modeled using square rate theory (SQRT) and macromolecular rate theory (MMRT) models. We found that the proportional nitrification rates at different temperatures varied among AOA to AOB ratios. Predicted by both models, an optimum temperature (Topt) for nitrification in AOA dominated soils was significantly higher than for soils where AOA and AOB abundances are within the same order of magnitude. Moreover, the change in heat capacity ( Δ C P ‡ ) associated with the temperature dependence of nitrification was positively correlated with Topt and significantly varied among the AOA to AOB ratios. The temperature ranges for NP decreased with increasing AOA abundance for both organic and inorganic fertilized soils. These results challenge the widely accepted approach of comparing NP rates in different soils at a fixed temperature. We conclude that a shift in AOA to AOB ratio in soils exhibits distinguished temperature-dependent characteristics that have an important impact on nitrification responses across the temperature gradient. The proposed approach benefits the accurate discernment of the true contribution of fertilized soils to nitrification for improvement of nitrogen management.

19.
Environ Sci Process Impacts ; 21(9): 1596-1608, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31414689

RESUMO

Soil nitrification responses to temperature have major implications for the global nitrogen cycle. Temperature sensitivity of soil nitrification has been modeled using several mathematical models, yet the extent to which model-generated thermodynamic parameters are accurate and sensitive in describing temperature sensitivity is unclear. In this study, we performed global sensitivity analysis to identify the key thermodynamic parameters that are most influential when simulating the temperature response of the soil nitrification potential (NP) across two different temperature gradients (4-40 °C and 20-45 °C) which are imposed upon sixteen different soils with square root growth (SQRT) and macromolecular rate theory (MMRT) models. We found that two thermodynamic parameters stand out as moderately to highly sensitive, and are uniquely identifiable in each model, regardless of the temperature range. The minimum and maximum measured temperatures seem to have no impact on the list of sensitive parameters but do influence the parameter ranges, especially for the SQRT model. However, parameters that control the minimum temperature and curvature of the NP response curve (Tmin and ΔC‡P) were found to have little to no sensitivity to SQRT and MMRT model outputs, respectively. We show that the parameter sensitivity and range of measured temperatures influence the complementary model's ability to describe the temperature sensitivity of soil nitrification. Our proposed framework enhances the accurate interpretation of existing thermodynamic parameters that explain the temperature sensitivity of soil biochemical processes, and provides methodological recommendations for future temperature sensitivity studies.


Assuntos
Modelos Teóricos , Nitrificação , Ciclo do Nitrogênio , Solo/química , Termodinâmica , Microbiologia do Solo , Temperatura
20.
Artigo em Inglês | MEDLINE | ID: mdl-28704958

RESUMO

This study presents an approach for obtaining realization sets of parameters for nitrogen removal in a pilot-scale waste stabilization pond (WSP) system. The proposed approach was designed for optimal parameterization, local sensitivity analysis, and global uncertainty analysis of a dynamic simulation model for the WSP by using the R software package Flexible Modeling Environment (R-FME) with the Markov chain Monte Carlo (MCMC) method. Additionally, generalized likelihood uncertainty estimation (GLUE) was integrated into the FME to evaluate the major parameters that affect the simulation outputs in the study WSP. Comprehensive modeling analysis was used to simulate and assess nine parameters and concentrations of ON-N, NH3-N and NO3-N. Results indicate that the integrated FME-GLUE-based model, with good Nash-Sutcliffe coefficients (0.53-0.69) and correlation coefficients (0.76-0.83), successfully simulates the concentrations of ON-N, NH3-N and NO3-N. Moreover, the Arrhenius constant was the only parameter sensitive to model performances of ON-N and NH3-N simulations. However, Nitrosomonas growth rate, the denitrification constant, and the maximum growth rate at 20 °C were sensitive to ON-N and NO3-N simulation, which was measured using global sensitivity.


Assuntos
Modelos Teóricos , Nitrogênio/análise , Eliminação de Resíduos Líquidos , Poluentes Químicos da Água/análise , Cadeias de Markov , Método de Monte Carlo , Nitrosomonas/crescimento & desenvolvimento , Lagoas , Software , Incerteza
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