Insight into the role of niche concept in deciphering the ecological drivers of MPs-associated bacterial communities in mangrove forest.

Myriad inherent and variable environmental features are controlling the assembly and succession of bacterial communities colonizing on mangrove microplastics (MPs). However, the mechanisms governing mangrove MPs-associated bacterial responses to environmental changes still remain unknown. Here, we assessed the dissimilarities of MPs-associated bacterial composition, diversity and functionality as well as quantified the niche variations of each taxon on plastispheres along river-mangrove-ocean and mangrove landward-to-seaward gradients in the Beibu Gulf, China, respectively. The bacterial richness and diversity as well as the niche breadth on mangrove sedimentary MPs dramatically decreased from landward to seaward regions. Characterizing the niche variations linked the difference of ecological drivers of MPs-associated bacterial populations and functions between river-mangrove-ocean (microplastic properties) and mangrove landward-to-seaward plastispheres (sediment physicochemical properties) to the trade-offs between selective stress exerted by inherent plastic substrates and microbial competitive stress imposed by environmental conditions. Notably, Rhodococcus erythropolis was predicted to be the generalist species and closely associated to biogeochemical cycles of mangrove plastispheres. Our work provides a reliable pathway for tackling the hidden mechanisms of environmental factors driving MPs-associated microbe from perspectives of niches and highlights the spatial dynamic variations of mangrove MPs-associated bacteria.

Comparison of the Glucocorticoid Receptor Binding and Agonist Activities of Typical Glucocorticoids: Insights into Their Endocrine Disrupting Effects.

Over the past decades, the synthetic glucocorticoids (GCs) have been widely used in clinical practice and animal husbandry. Given the health hazard of these toxic residues in food, it is necessary to explore the detailed interaction mechanisms of typical GCs and their main target glucocorticoid receptor (GR). Hence, this work compared the GR binding and agonist activities of typical GCs. Fluorescence polarization assay showed that these GCs were potent ligands of GR. Their GR binding affinities were in the order of methylprednisolone>betamethasone≈prednisolone>dexamethasone, with IC50 values of 1.67, 2.94, 2.95, and 5.58 nM. Additionally, the limits of detection of dexamethasone, betamethasone, prednisolone, and methylprednisolone were 0.32, 0.14, 0.19, and 0.09 µg/kg in fluorescence polarization assay. Reporter gene assay showed that these GCs induced GR transactivation in a dose-dependent manner, confirming their GR agonist activities. Among which, dexamethasone at the concentration of 100 nM produced a maximal induction of more than 11-fold over the blank control. Molecular docking and molecular dynamics simulations suggested that hydrogen-bonding and hydrophobic interactions played an important role in stabilizing the GC-GR-LBD complexes. In summary, this work might help to understand the GR-mediated endocrine disrupting effects of typical GCs.

In vitro, in vivo, and in silico evaluation of the glucocorticoid receptor antagonist activity of 3,6-dibromocarbazole.

3,6-Dibromocarbazole is a novel environmental contaminant which is currently detected in several environmental media worldwide. This work aims to investigate the anti-glucocorticoid potency and endocrine disrupting effects of 3,6-dibromocarbazole. In vitro experiments indicated that 3,6-dibromocarbazole possessed glucocorticoid receptor (GR) antagonistic activity and inhibited dexamethasone-induced GR nuclear translocation. 3,6-Dibromocarbazole reduced the expression levels of glucocorticoid responsive genes including glucose-6-phosphatase (G6Pase), phosphoenolpyruvate carboxykinase (PEPCK), fatty acid synthase (FAS), and tyrosine aminotransferase (TAT), and further disrupted the protein expression of two key enzymes PEPCK and FAS in gluconeogenesis. In vivo experiments showed that 3,6-dibromocarbazole induced abnormal development of zebrafish embryos and disrupted the major neurohormones involved in activation of hypothalamic-pituitary-adrenocortical (HPA) axis in zebrafish larvae. The results of molecular docking and molecular dynamics simulation contributed to explain the antagonistic effect of 3,6-dibromocarbazole. Taken together, this work identified 3,6-dibromocarbazole as a GR antagonist, which might exert endocrine disrupting effects by interfering the pathway of gluconeogenesis.

Distinct microplastics abundance variation in root-associated sediments revealed the underestimation of mangrove microplastics pollution.

Mangrove sediment is acknowledged as the critical sink of microplastics (MPs). However, the potential effect of mangrove root systems on the MPs migration in sediment remains largely unknown. Here, our study characterized the spatial distribution of MPs trapped in root hair, rhizosphere, and non-rhizosphere zones, and analyzed their correlations with physicochemical properties of sediments. The significantly increased MPs abundances toward root systems shed light on the distinct effect on the migration of MPs exerted by mangrove root systems. Partial least squares path modeling (PLS-PM) analysis revealed that pore water content and pH influenced the abundances of different MP characteristics (shape, color, size, and type) and further promoted the accumulation of MPs toward the root systems. In different mangrove areas from landward to seaward, other sediment properties (median grain size, clay content, and salinity) also controlled MP distribution. Additionally, smaller-sized MPs (<1000 µm) were more easily transported to the root systems. Our study emphasizes the importance of considering root systems effect when investigating the mechanisms of MPs distribution and migration in mangrove sediments.

Selective enrichments for color microplastics loading of marine lipophilic phycotoxins.

Microplastics (MPs) and marine lipophilic phycotoxins (MLPs) are two classes of emerging contaminants. Together, they may exacerbate the negative impacts on nearshore marine ecosystems. Herein, the loading of 14 representative MLPs, closely related to toxin-producing algae, on MPs and their relations with colorful MPs have been explored for the first time based on both field and lab data. The objectives of our study are to explore the roles of multiple factors (waterborne MLPs and MP characteristics) in the loading of MLPs by MPs with the applications of various statistical means, and to further explore the role of the color of MP in the loading of specific MLPs through lab simulation experiments. Our results demonstrated that MPs color determined the loading of some specific MLPs on MPs and green MPs can load much more than other colorful fractions (p < 0.05). These interesting phenomena illustrated that the color effects on the loading processes of MLPs on MPs are a dynamic process, and it can be well explained by the shading effect of MP color, which may affect the growth and metabolism of the attached toxic-producing algae on MPs and hence the production of specific MLPs. Furthermore, loading of MLPs on MPs can be considered as the comprehensive physicochemical and biological processes. Our results caution us that special attention should be paid to explore the real-time dynamic color shading effects on all kinds of bio-secreted contaminants loading on MPs, and highlight the necessary to comprehensive investigate the interaction between biota, organic contaminants and MPs.

Identifying the electricity-saving driving behaviors of electric bus based on trip-level electricity consumption: a machine learning approach.

Electric buses (EBs) are gaining popularity worldwide as a more sustainable and eco-friendly alternative to diesel buses (DBs). Electricity-saving driving plays a crucial role in minimizing an EB's energy consumption, subsequently leading to an extended driving range. This study proposes a machine learning-based framework for identifying electricity-saving EB driving behaviors during various driving stages, including running on road segments, entering bus stops/intersections, and exiting bus stops/intersections. The proposed random forest (RF) model effectively evaluates the energy consumption level using EB drivers' historical driving data under different scenarios. Specifically, the electricity consumption factor (ECF), as the evaluation index, is divided into three categories to determine the implicit relationship between driving behavior and energy consumption. The results indicate that the classification accuracy of RF models surpasses 90%, which highlights the effectiveness in accurately identifying energy-efficient EB driving behaviors. In addition, the Shapley additive explanations (SHAP) and partial dependency plots (PDPs) are utilized to visualize and interpret the results of RF models. A speed interval of 30-40 km/h is identified as the most energy-efficient range for EB running on a road segment. Findings from this study can be applied to targeted optimization of electricity-saving driving strategies in different driving scenarios to improve the overall efficiency and sustainability of the transportation system.

Dietary regulation of peroxisome proliferator-activated receptors in metabolic syndrome.

BACKGROUND: Peroxisome proliferator-activated receptors (PPARs) are a class of ligand-activated nuclear transcription factors, members of the type nuclear receptor superfamily, with three subtypes, namely PPARα, PPARß/δ, and PPARγ, which play a key role in the metabolic syndrome. In the past decades, a large number of studies have shown that natural products can act by regulating metabolic pathways mediated by PPARs. PURPOSE: This work summarizes the physiological importance and clinical significance of PPARs and reviews the experimental evidence that natural products mediate metabolic syndrome via PPARs. METHODS: This study reviews relevant literature on clinical trials, epidemiology, animals, and cell cultures published in NCBI PubMed, Scopus, Web of Science, Google Scholar, and other databases from 2001 to October 2022. Search keywords were "natural product" OR "botanical" OR "phytochemical" AND "PPAR" as well as free text words. RESULTS: The modulatory involvement of PPARs in the metabolic syndrome has been supported by prior research. It has been observed that many natural products can treat metabolic syndrome by altering PPARs. The majority of currently described natural compounds are mild PPAR-selective agonists with therapeutic effects that are equivalent to synthetic medicines but less harmful adverse effects. CONCLUSION: PPAR agonists can be combined with natural products to treat and prevent metabolic syndrome. Further human investigations are required because it is unknown how natural products cause harm and how they might have negative impacts.

Role of mangrove forest in interception of microplastics (MPs): Challenges, progress, and prospects.

Mangroves receive microplastics (MPs) from terrestrial, marine and atmospheric sources, acting as a huge filter for environmental MPs between land and sea. Due to the high primary production and complex hydrodynamic conditions in mangroves, MPs are extensively intercepted in various ways while flowing through mangroves, leading to a long-standing but fiercely increasing MPs accumulation. However, current researches mainly focused on the occurrence, source and fate of MPs pollution in mangroves, ignoring the role of mangrove forests in the interception of MPs. Our study firstly demonstrates that mangrove ecosystems have significantly greater MPs interception capacity than their surrounding environments. Then, the current status of studies related to the interception of MPs in mangrove ecosystems is comprehensively reviewed, with the main focus on the interception process and mechanisms. At last, the most pressing shortcomings of current research are highlighted regarding the intercepted flux, interception mechanisms, retention time and ecological risks of MPs in mangrove ecosystems and the relevant future perspectives are provided. This review is expected to emphasize the critical role of mangrove forests in the interception of MPs and provide the foundational knowledge for evaluating the MPs interception effect of mangrove forests globally.

Characterization of tangeretin as an activator of nuclear factor erythroid 2-related factor 2/antioxidant response element pathway in HEK293T cells.

Numerous studies have reported that tangeretin is a polymethoxylated flavone with a variety of biological activates, but little research has been done on the antioxidant mechanism of tangeretin. Hence, we investigated the effect of tangeretin on the nuclear factor erythroid 2-related factor 2 (Nrf2)/antioxidant response element (ARE) pathway and its potential molecular mechanisms by in vitro and in silico research. The results of molecular docking suggested that tangeretin bound at the top of the central pore of Kelch-like ECH-associated protein 1 (Keap1) Kelch domain, and the hydrophobic and hydrogen bond interactions contributed to their stable binding. Herein, the regulation of Nrf2-ARE pathway by tangeretin was explored in the human embryonic kidney cell line HEK293T, which is relatively easy to be transfected. Upon binding to tangeretin, Nrf2 translocated to the nucleus of HEK293T cells, which in turn activated the Nrf2-ARE pathway. Luciferase reporter gene analysis showed that tangeretin significantly induced ARE-mediated transcriptional activation. Real-time PCR and Western blot assays showed that tangeretin induced the gene and protein expressions of Nrf2-mediated targets, including heme oxygenase 1 (HO-1), nicotinamide adenine dinucleotide phosphate (NADPH) quinone dehydrogenase 1 (NQO1), and glutamate-cysteine ligase (GCLM). In addition, tangeretin could effectively scavenge 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radicals. In summary, tangeretin may be a potential antioxidant via activating the Nrf2-ARE pathway.

Quercetin ameliorates hepatic fat accumulation in high-fat diet-induced obese mice via PPARs.

As a natural pigment in food, quercetin possesses multiple biological activities and plays a crucial role in regulating metabolic syndrome. Herein, we aim to explore the potential mechanism of quercetin to ameliorate hepatic fat accumulation. In vivo experiments showed that quercetin significantly relieved inflammation response by decreasing the serum TNF-α and IL-6 levels and also improved high-fat diet-induced hepatic steatosis without other organ injuries. Quercetin can effectively reduce lipid aggregation and down-regulate the protein expression of PCK1 in HepG2 cells induced by oleic acid and palmitic acid, indicating that inhibiting gluconeogenesis leads to hepatic fat accumulation reduction. Furthermore, molecular docking results suggested that quercetin can bind to both PPARα and PPARγ, with an even more potent binding affinity than indeglitazar, a pan-agonist of PPARs. In conclusion, quercetin may regulate gluconeogenesis to ameliorate hepatic fat accumulation via targeting PPARα/γ.

Riverine microplastics derived from mulch film in Hainan Island: Occurrence, source and fate.

Mulch film (MF) residues is an important source of microplastics (MPs) in farmland, but its transportation risk to the wider environment was still unknown. Some researches have pursued the sources of MPs found in exorheic rivers. Even so, a systematic study depicting the occurrence, source and fate of microplastics derived from mulch films (MPMF), the crucial component of MPs in farmlands, in exorheic rivers still lacking. Here, the combination of UV-Vis Raman spectroscopy and X-ray photoelectron spectroscopy (XPS) was used to identify the full-size MPMF (1-5000 µm) in field sediment samples collected by single-diagonal systematic sampling. This study verified that MPMF, a polyethylene-matrix composite doped with additives, contributed a considerable part of MPs detected in upstream farmland soil and riverine sediments, and even had an abundance of 38 ± 11 items/kg to 82 ± 15 items/kg, accounting for 9.0%-13.7% of the total MPs in estuary sediments. Notably, upstream farmland was identified to the main source of the riverine MPMF by partial least square path modeling (PLS-PM), contributing to 94.7% of MPMF in riverside sediments and 85.0% of MPMF in estuary sediments. Our study first demonstrates that MPMF constitutes a non-negligible component of MPs in estuarine sediments and underlines the urgency of strengthening the management of MPs pollution in drainage areas with a high agricultural intensity.

Distribution and retention of microplastics in plantation mangrove forest sediments.

Microplastics (MPs) in ocean tides can be effectively intercepted by mangroves, especially sediments, which are considered to be effective sinks. However, the retention of plantation mangrove forests on MPs is still unclear. In this study, the spatial distribution and its implication factors of MPs in surface sediments of plantation mangrove forests were investigated for the first time. In plantation forests, MPs were detected with abundances ranging from 67 ± 21 to 203 ± 25 items/kg, and plantation forests were significantly lower than natural forests at the CJ sampling site (p < 0.05). Plantation forests had fewer fibrous MPs than natural forests (p < 0.05). Furthermore, the MPs abundance showed strong linear relationships with the sand content (p = 0.002, R2 = 0.86) and Aegiceras corniculata biomass (p = 0.001, R2 = 0.84). Partial least squares path modeling analysis (PLS-PM) indicated that these two factors influenced MPs abundance by retaining MPs with fibrous, fragmented, denser and larger-sized characteristics. Our results revealed the differences in MPs abundance and characteristics between plantation and natural mangrove forests, and it is necessary to monitor MPs pollution to provide significant guidance for the restoration of constructed wetlands.

Effective and Acceptable Eco-Driving Guidance for Human-Driving Vehicles: A Review.

Eco-driving guidance refers to courses, warnings, or suggestions provided to human drivers to improve driving behaviour to enable less energy use and emissions. This paper reviews existing eco-driving guidance studies and identifies challenges to tackle in the future. We summarize two categories of current guidance systems, static and dynamic, distinguished by whether real-world driving records are used to generate behaviour guidance or not. We find that influencing factors, such as the content of suggestions, the display methods, and drivers' socio-demographic characteristics, have varied effects on the guidance results across studies. Drivers are reported to have basic eco-driving knowledge, while the question of how to motivate the acceptance and practice of such behaviour, especially in the long term, is overlooked. Adaptive driving suggestions based on drivers' individual habits can improve the effectiveness and acceptance while this field is under investigation. In-vehicle assistance presents potential safety issues, and visualized in-vehicle assistance is reported to be most distractive. Given existing studies focusing on the operational level, a common agreement on the guidance design and associated influencing factors has yet to be reached. Research on the systematic and tactical design of eco-driving guidance and in-vehicle interaction is advised.

Impact of vehicle type, tyre feature and driving behaviour on tyre wear under real-world driving conditions.

Tyre wear generates not only large pieces of microplastics but also airborne particle emissions, which have attracted considerable attention due to their adverse impacts on the environment, human health, and the water system. However, the study on tyre wear is scarce in real-world driving conditions. In the present study, the left-front and left-rear tyre wear in terms of volume lost in mm3 of 76 taxi cars was measured about every three months. This study covered 22 months from September 2019 to June 2021 and included more than 500 measurements in total. Some of the data was used to evaluate the effects of vehicle type and tyre type on tyre wear. In addition, a machine learning method (i.e., Extreme gradient boosting (XGBoost)) was used to probe the effect of driving behaviour on tyre wear by monitoring real-time driving behaviour. The current statistical results showed that, on average, the tyre wear was 72 mg veh-1 km-1 for a hybrid car and 53 mg veh-1 km-1 for a conventional internal combustion engine car. The average tyre wear measured for a taxi vehicle configuration featuring winter tyres was 160 mg veh-1 km-1, which was 1.4 and 3.0 times as much as those with all-season tyres and summer tyres, respectively. The wear rate of left-front tyres was 1.7 times higher than that of left-rear tyres. The XGBoost results indicated that compared to driving behaviour, tyre type and tyre position had more important effects on tyre wear. Among driving behaviours, braking and accelerating events presented the most considerable impact on tyre wear, followed by cornering manoeuvres and driving speed. Thus, it seems that limiting harsh braking and acceleration has the potential to reduce tyre wear significantly.

Quantifying the Impact of Alternative Bus Stop Platforms on Vehicle Emissions and Individual Pollution Exposure at Bus Stops.

Due to stop-and-go events, bus stops are often treated as "hot spots" for air pollution. The design of bus stops should be optimized to reduce emissions and exposure for transit commuters. The objective of this study was to analyze the impact of bus stop platform types on vehicle emissions and individual pollution exposure. Second-by-second emissions data were first collected from one bus using a portable emission measurement system (PEMS). Microscopic traffic simulation was then used to estimate overall traffic emissions under six scenarios with different bus stop settings. Numerical simulation of pollutant dispersion was also conducted to calculate individual pollution exposure at bus stops. The results of PEMS tests showed no significant differences between bus emissions generated near two different types of stops. However, the effect of platform types on overall traffic emissions was revealed using traffic simulation. The results demonstrated that bus bays reduced the emissions of other heavy-duty vehicles. However, bus bays were not always effective during rush hours. The study also highlighted the importance of the location of bus stops, the number of bus lines, and the length of the platform, in addition to dynamic characteristics of traffic flows in the design of bus stop platforms. Bus stop platforms also affected individuals' exposure due to the changes in the pollutant flow field. The passenger's exposure at one bus stop was influenced by both the platform type and standing location. Results suggested that in a condition with a wind direction perpendicular to the bus stop shelter, the total exposure level to CO was lower at the bus bay stop if a passenger stood at the upstream of the station platform. However, the exposure was less at the downstream of the curbside bus stop.

Mangrove forest: An important coastal ecosystem to intercept river microplastics.

The research on transportation of river microplastics (MPs) mainly focuses on the estimations of the total contents of river MPs entering the ocean, while the related transportation processes and influence factors were still largely unknown. In our study, the role of mangrove forest, a special tropical ecosystem in the estuary, on the transportations of MPs from rivers to ocean was explored. Except for the ND river with the absence of mangrove forest, the MPs collected from the water sample of the river upstream were much higher than their corresponding downstream (p < 0.05), with the interception rate of riverine MPs by mangrove forests ranging from 12.86% to 56% in dry season and 10.57%-42% in rainy season. The MPs with the characteristics of high density, larger size and regular shape were more easily intercepted. Furthermore, the combined effects of ecological indicators, the properties of mangrove and the hydrodynamic factors jointly determined the interception rates of MPs. This study provides a new perspective and data support for quantifying mangrove forests intercepting MPs in rivers as a factor of MPs retention in global rivers.

Characterization of diphenyl phthalate as an agonist for estrogen receptor: an in vitro and in silico study.

Phthalate esters (PAEs) are important pollutants in the environment, which can interfere with the endocrine system by mimicking estrogen. However, limited information is available on modulating the estrogen receptor (ER) of five PAEs including di (2-ethylhexyl) phthalate (DEHP), diisononyl phthalate (DINP), benzyl butyl phthalate (BBP), diphenyl phthalate (DPhP) and dicyclohexyl phthalate (DCHP). This study evaluated the agonistic effects of PAEs on human ER. The cytotoxicity assay showed that there were a significant inhibition of the cell proliferation with treatment of five PAEs. Moreover, DPhP does-dependently enhanced ER-mediated transcriptional activity in the reporter gene assay. The increased expression of estrogen-responsive genes (TFF1, CTSD, and GREB1) was also observed in MCF-7 cells treated with DPhP. The result of molecular docking showed that DPhP tended to bind to the agonist conformation of ER compared with the antagonist conformation of ER, demonstrating its agonist characteristic that has been confirmed in the reporter gene assay. Thus, we found that DPhP may be evaluated as an ER agonist in vitro and it can interfere with the normal function of human ER.

Comparison of NOx and PN emissions between Euro 6 petrol and diesel passenger cars under real-world driving conditions.

With emission standards becoming stricter, nitrogen oxides (NOx) and particle number (PN) emissions are the main concerns of modern passenger cars, especially for the real-world driving. In this paper, two direct injection (DI) petrol passenger cars and a diesel passenger car are tested on the same routes, driven by the same driver. Instantaneous NOx and PN emissions are monitored by a portable emission measurement system (PEMS) in the tests. During the real-world driving, the exhaust temperatures of the two petrol cars are sufficiently high to ensure high efficiency of three-way catalysts (TWCs). On the other hand, the exhaust temperatures of the diesel car in some sections of the route are lower than the crucial light-off temperature of the selective catalytic reduction (SCR) below which its effectiveness in NOx reduction would be much affected. NOx and PN concentrations are low during motorway driving for the petrol passenger car equipped with a gasoline particulate filter (GPF); however, they are high and change frequently in the whole journey for the petrol passenger car without a GPF. NOx emission factors are quite low over most of the driving sections for the diesel car, but some significant high peaks are observed in the acceleration process. NOx emission distributions over speed and acceleration are similar for both petrol cars; and they differ significantly from the diesel counterpart. Particle size from the diesel car is the largest, followed by the petrol car with a GPF.

Identification of dicyclohexyl phthalate as a glucocorticoid receptor antagonist by molecular docking and multiple in vitro methods.

The potential activities of phthalate esters (PAEs) that interfere with the endocrine system have been focused recently. However, information on modulating the glucocorticoid receptor (GR) of PAEs is scarce. Our aim was to evaluate the agonistic / antagonistic properties of PAEs on human GR. Luciferase reporter gene assay revealed that the tested chemicals displayed no agonistic effects but dicyclohexyl phthalate (DCHP) exerted antagonistic activity in a dose-responsive manner for GR in HeLa cells. The effects of DCHP on dexamethasone (DEX)-induced GR nuclear translocation and gene expression of glucocorticoid-responsive gene expression (G6Pase, PEPCK, FAS, GILZ and MKP-1), as well as protein expression of G6Pase and PEPCK were further examined by RT-qPCR and western blot analysis. DCHP antagonized DEX-induced GR nuclear translocation and suppressed gene expression in both mRNA and protein levels. Furthermore, the results of molecular docking and molecular dynamics simulation showed that DCHP could bind to GR and exhibited potential regulation on this target protein. Collectively, we demonstrate that DCHP may act as a GR antagonist in vitro and is considered to exert endocrine effects via human GR.

Real-world emissions of construction mobile machines and comparison to a non-road emission model.

This study implemented real-world tests in Nanjing, China for measuring emission factors (EFs) of air pollutants, including Carbon Monoxide (CO), Hydrocarbon (HC), Nitrogen Oxides (NOx), and Particulate Matter (PM) from ten construction machines in three operational modes (idling, moving, and working) with a Portable Emission Measurement System. The idling mode shows the least variation of EFs, and its average CO EFs can be higher than the moving and working modes by 43% and 34%, respectively. The working mode generates the highest emission for all other pollutants with the highest variation. The EFs suggested by the Guide (an official guidebook for developing emission inventory in China) are in general lower than the measured EFs, and the gap becomes larger for older machines. The EFs of CO, NOx, and PM of China Stage II machines are 24%, 120%, and 66% higher than those of the Guide, respectively. The differences go up as high as 126%, 1066%, and 559% for China Stage I machines, indicating the upgrade of engine technology from Stage I to Stage II, as well as the effect of machine deterioration. The result of this study reveals the effectiveness of stringent emission standards in controlling emissions from construction machines. High emissions from older machines emphasize the importance of a more rigorous machine replacement policy and a regulated maintenance strategy. The result also stresses the need to update the Guide with differentiated activity modes, region variations, and machine deterioration effects.