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At least 973 records · Page 54

Near-infrared laser irradiation improves the development of mouse pre-implantation embryos

The aim of the present study was to assess the effects of near-infrared laser irradiation on the in vitro development of mouse embryos. Female ICR mice were superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin (hCG), and mated with male mice. Two-cell stage embryos were collected 40 h after administering hCG and cultured in M16 medium. Two-cell embryos (0 h after culture), 8-cell embryos (approx. 30 h after culture), morula (approx. 48 h after culture), and blastocysts (approx. 73 h after culture) were irradiated at 904 nm for 60 s. These embryos were cultured in a time-lapse monitoring system and the timing of blastocyst hatching was evaluated. Some of the irradiated blastocysts were transferred to the uterine horns of pseudopregnant recipients immediately after irradiation. Pregnancy rates, and offspring growth and fertility, were evaluated. Near-infrared laser irradiation increased the speed of in vitro mouse embryo development. In irradiated blastocysts, hatching was faster than in control (non-irradiated) blastocysts (18.4 vs. 28.2 h, P < 0.05). When 195 irradiated blastocysts were transferred to 18 pseudopregnant mice, all became pregnant and 92 (47.2%) normal-looking pups were born alive. When 182 control blastocysts were transferred to 17 pseudopregnant mice, 14 (82.4%) became pregnant and 54 (29.7%) normal-looking pups were born alive. The growth trajectories (up to 5 weeks) of offspring from irradiated blastocysts were similar to those from control blastocysts. Second generation offspring from transplanted animals were all fertile. These results indicate that near-infrared laser irradiation improves the quality of mouse embryo development in vitro, and increases the live birth rate without affecting the normality of the offspring. Thus, the near-infrared laser method may enhance the quality of embryos and contribute to improvements in reproductive technologies in mammals. - Highlights: • Irradiation of blastocysts with a near-infrared laser improves embryo development. • Irradiation of blastocysts increases the live birth rate after embryo transfer. • Irradiation of blastocysts did not affect the normality of the pups. • Near-infrared laser irradiation may be useful to enhance the quality of embryos. • This study may contribute to improvements in reproductive technologies in mammals.

60 APPLIED LIFE SCIENCES↗

Newly developed selective immunoinactivation assay revealed reduction in adipose triglyceride lipase activity in peripheral leucocytes from patients with idiopathic triglyceride deposit cardiomyovasculopathy

Highlights: • Development of selective immunoinactivation assay for ATGL activity in leucocytes. • ATGL activity was markedly reduced in idiopathic TGCV without ATGL mutations. • ATGL in leucocytes may be an important biomarker for the diagnosis of TGCV. • Selective immunoinactivation assay provides insights into pathogenesis for TGCV. Triglyceride deposit cardiomyovasculopathy (TGCV) is a rare and newly identified disease among patients requiring cardiac transplantation. TGCV is characterized by cardiomyocyte steatosis and triglyceride (TG)-deposit atherosclerosis, resulting from the abnormal intracellular metabolism of TG. TGCV is classified into primary and idiopathic types. Primary TGCV carries ultra-rare genetic mutations in the adipose triglyceride lipase (ATGL), a rate-liming enzyme that hydrolyzes intracellular TG in adipose and non-adipose tissues. Idiopathic TGCV, first identified among autopsied individuals with diabetes mellitus (DM) with severe heart diseases, shows no ATGL mutations and its causes and underlying mechanisms are still unknown. TGCV is difficult to diagnose in daily clinics, thereby demanding feasible diagnostic procedures. We aimed to develop an assay to measure ATGL activity using peripheral leucocytes. Human his6-ATGL was expressed in COS1 cells, purified to homogeneity, and used to raise a polyclonal antibody neutralizing TG-hydrolyzing activity of ATGL. We developed a selective immunoinactivation assay (SIIA) for the quantitation of ATGL activity in cell lysates of leucocytes by the antibody neutralizing ATGL activities. ATGL activity was measured in 13 idiopathic TGCV patients, with two patients with primary TGCV as the negative control. Healthy (non-DM) and DM controls without heart diseases were also subjected. The developed SIIA assay revealed significant reduction in ATGL activity in leucocytes from patients with idiopathic TGCV who did not carry ATGL mutations as compared with non-DM and DM controls. Thus, ATGL in leucocytes may be an important biomarker for the diagnosis of TGCV and our assay may provide insights into pathophysiology and elucidate the underlying mechanism of TGCV and related disorders.

60 APPLIED LIFE SCIENCES↗

Distinct expression patterns of Flk1 and Flt1 in the coronary vascular system during development and after myocardial infarction

Highlights: • Flk1 is expressed in ECs with an epicardial-to-endocardial transmural gradient. • Flt1 is homogeneously expressed in ECs in neonatal and adult mouse hearts. • Flk1 is transiently induced but Flt1 is maintained in coronary vessels after MI. • The kinetics of Flk1 and Flt1 after MI recapitulates that in cardiac development. The coronary vascular system is critical for myocardial growth and cardiomyocyte survival. However, the molecular mechanism regulating coronary angiogenesis remains elusive. Vascular endothelial growth factor (VEGF) regulates angiogenesis by binding to the specific receptors Flk1 and Flt1, which results in different functions. Despite the importance of Flk1 and Flt1, their expression in the coronary vasculature remains largely unknown due to the lack of appropriate antibodies for immunostaining. Here, we analyzed multiple reporter mice including Flk1-GFP BAC transgenic (Tg), Flk1-LacZ knock-in, Flt1-DsRed BAC Tg, and Flk1-GFP/Flt1-DsRed double Tg animals to determine expression patterns in mouse hearts during cardiac growth and after myocardial infarction (MI). We found that Flk1 was expressed in endothelial cells (ECs) with a pattern of epicardial-to-endocardial transmural gradients in the neonatal mouse ventricle, which was downregulated in adult coronary vessels with development. In contrast, Flt1 was homogeneously expressed in the ECs of neonatal mouse hearts and expression was maintained until adulthood. After MI, expression of both Flk1 and Flt1 was induced in the regenerating coronary vessels at day 7. Intriguingly, Flk1 expression was downregulated thereafter, whereas Flt1 expression was maintained in the newly formed coronary vessels until 30 days post-MI, recapitulating their expression kinetics during development. This is the first report demonstrating the spatiotemporal expression patterns of Flk1 and Flt1 in the coronary vascular system during development and after MI; thus, this study suggests that these factors have distinct and important functions in coronary angiogenesis.

60 APPLIED LIFE SCIENCES↗

Harmine promotes molar root development via SMAD1/5/8 phosphorylation

Tooth and bone are major tissues involved in physiological calcification in the body, and they use similar molecular pathways for development, homeostasis, and regeneration. Harmine (HMN) is a natural small compound that stimulates osteoblast differentiation in vitro and in vivo. Here we examined the biological effect of HMN on the postnatal development of molar tooth roots and periodontal tissues. HMN supported the formation of tooth roots and periodontal tissues in developing tooth germs. In tooth germ organ culture, HMN promoted the elongation of Hertwig's epithelial root sheath (HERS) and stimulated cell proliferation in HERS and dental follicle-derived tissues, including dental papillae and dental follicles. HMN stimulated cell proliferation and cell movement of HERS-derived cells without mesenchymal cells in vitro and directly induced the phosphorylation of SMAD1/5/8 protein in HERS-derived cells. Our results indicated that HMN was the first natural small compound to stimulate postnatal development of tooth germs.

60 APPLIED LIFE SCIENCES↗

Mice overexpressing latent matrix metalloproteinase-2 develop lung emphysema after short-term exposure to cigarette smoke extract

Highlights: • Protease/anti-protease imbalance is an important event in the pathogenesis of lung emphysema. • Long time exposure to cigarette smoke is generally required to induce mouse model of lung emphysema. • Mice overexpressing matrix metalloproteinase-2 develop lung emphysema after short exposure to cigarette smoke extract. Chronic obstructive pulmonary disease is the major growing cause of mortality and morbidity worldwide, and it is going to become the third most common cause of death by 2020. Chronic obstructive pulmonary disease is pathologically characterized by lung emphysema and small airway inflammation. Animal models are very important to get insights into the disease pathogenesis but current models of chronic obstructive pulmonary disease take a long time to develop. The need of a new model is compelling. In the present study we focus on the role of matrix metalloproteinases in the pathogenesis of chronic obstructive pulmonary disease and hypothesized that lung overexpression of latent matrix metalloproteinases-2 would allow the development of emphysema after short-term exposure to cigarette smoke extract inhalation. Human latent matrix metalloproteinases-2 transgenic mouse expressing high level of the protein in the lungs and wild type mouse were exposed to aerosolized cigarette smoke extract for two weeks. Transgenic mice showed significant lung emphysematous changes, increased infiltration of inflammatory cells and enhanced lung concentrations of inflammatory cytokines in the lungs compared to their wild type counterparts after inhalation of cigarette smoke extract. This novel mouse model will be a very useful tool for evaluating the mechanistic pathways and for development of novel therapies in cigarette smoke-associated lung emphysema.

60 APPLIED LIFE SCIENCES↗

Loss of RapC causes defects in cytokinesis, cell migration, and multicellular development of Dictyostelium

Highlights: • We characterized the functions of RapC, an isoform of Rap1 small GTPase. • Loss of RapC causes defects in cytokinesis, cell migration, and development. • RapC plays a negative role in cell spreading and cell adhesion. • RapC is involved in tip formation from mounds during multicellular development. • This contributes to the understanding of Ras-mediated cellular processes. The small GTPase Ras proteins are involved in diverse cellular processes. We investigated the functions of RapC, one of 15 Ras subfamily GTPases in Dictyostelium. Loss of RapC resulted in a spread shape of cells; severe defects in cytokinesis leading to multinucleation; decrease of migration speed in chemoattractant-mediated cell migration, likely through increased cell adhesion; and aberrations in multicellular development producing abnormal multiple tips from one mound and multi-branched developmental structures. Defects in cells lacking RapC were rescued by expressing GFP-RapC in rapC null cells. Our results demonstrate that RapC, despite its high sequence homology with Rap1, plays a negative role in cell spreading and cell adhesion, in contrast to Rap1, which is a key regulator of cell adhesion and cytoskeleton rearrangement. In addition, RapC appears to have a unique function in multicellular development and is involved in tip formation from mounds. This study contributes to the understanding of Ras-mediated cellular processes.

60 APPLIED LIFE SCIENCES↗

RBM14 is indispensable for pluripotency maintenance and mesoderm development of mouse embryonic stem cells

Highlights: • RBM14 knockout mouse embryonic stem cells were successfully generated. • RBM14 depletion altered gene expression profiles in mouse embryonic stem cells. • RBM14 depletion impeded mesoderm development in vitro. • RBM14 depletion impeded mesoderm development in vivo. The pluripotency of embryonic stem cells (ESCs) is maintained by core pluripotency transcription factors, cofactors and several signaling pathways. RBM14 is a component of the para-speckle complex, which has been implicated in multiple important biological processes. The role of RBM14 in ESCs and lineage differentiation remains to be elucidated. In the present study, we provided evidence that RBM14 plays important roles in maintaining pluripotency and in the early differentiation of ESCs. RBM14 was demonstrated to be expressed in mouse embryonic stem cells (mESCs) and localized in the nucleus. RBM14 expression was depleted in mESCs using clustered regularly interspaced short palindromic repeats (CRISPR) technology. Our results also showed that RBM14 depletion altered the gene expression profiles of mESCs. In particular, pluripotency-associated genes and genes involved in the Wnt and TGF-β signaling pathways were downregulated in RBM14 knockout mESCs. Furthermore, RBM14 was found to be essential for mesoderm development in vitro and in vivo. The specific effects of RBM14 depletion were verified by conducting a rescue experiment. Our findings demonstrated that RBM14 not only plays an important role in maintaining the pluripotency of mESCs but is also indispensable for mesoderm development.

60 APPLIED LIFE SCIENCES↗

FOXP4-AS1 participates in the development and progression of osteosarcoma by downregulating LATS1 via binding to LSD1 and EZH2

Highlights: • We first confirmed that FOXP4-AS1 is overexpressed in OS, and is the independent risk factor in OS prognosis. • We further clarified that Upregulated FOXP4-AS1 promotes the proliferation, migration and cell cycle, but inhibits apoptosis of OS cells. • Finally, we demonstrated that FOXP4-AS1 participates in the development and progression of OS by downregulating LATS1. Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. LncRNA has been confirmed to participate in a variety of cancers. The purpose of this study was to explore the effect of FOXP4-AS1 on the development of osteosarcoma (OS) and its underlying mechanism. FOXP4-AS1 expressions in 60 OS tissues and paracancerous tissues were detected by qRT-PCR (quantitative real-time polymerase chain reaction). We confirmed that FOXP4-AS1 was overexpressed in OS tissues than that of paracancerous tissues. The disease-free survival and overall survival of OS patients were not correlated with age, gender and tumor location, but remarkably correlated with FOXP4-AS1 expression, tumor size and lung metastasis. For in vitro experiments, MG63 cells expressed a higher expression of FOXP4-AS1, whereas U2OS cells expressed a lower expression, which were selected for the following studies. Overexpressed FOXP4-AS1 led to enhanced proliferation, migration and invasion, shortened G0/G1 phase, as well as inhibited cell cycle. Knockdown of FOXP4-AS1 in MG63 cells obtained the opposite results. Furthermore, RIP assay indicated that FOXP4-AS1 could inhibit LATS1 expression by binding to LSD1 and EZH2, so as to participate in OS development. In conclusion, these results revealed that FOXP4-AS1 is overexpressed in OS, and is the independent risk factor in OS prognosis. Upregulated FOXP4-AS1 promotes the proliferation, migration and cell cycle, but inhibits apoptosis of OS cells. Furthermore, FOXP4-AS1 participates in the development and progression of OS by downregulating LATS1 via binding to LSD1 and EZH2.

60 APPLIED LIFE SCIENCES↗

The zinc-finger transcription factor BcMF20 and its orthologs in Cruciferae which are required for pollen development

Highlights: • BcMF20 was a three-fingered protein and close to the four-fingered proteins of petunia. • The inhibition of BcMF20 led to fewer and lower rate in pollen germination. • The inhibition of BcMF20 led to lower rate in fruit setting. • BcMF20 may work in genetic pathways to act on proliferation of tapetal cells. • BcMF20 helps keep the normal development of pollens. Brassica campestris Male Fertility 20 (BcMF20) is a typical zinc-finger transcription factor that was previously isolated from flower buds of Chinese cabbage (Brassica campestris ssp. chinensis). By applying expression pattern analysis, it can be known that BcMF20 was specifically and strongly expressed in tapetum and pollen, beginning from the uninucleate stage, and was maintained during the mature-pollen stage. As BcMF20 was highly conserved in Cruciferae, it can be indicated that this zinc-finger transcription factor is important during the growth of Cruciferae. In this study, 12 C2H2-type zinc-finger TFs which shared high homology with BcMF20 were found from NCBI via BLAST. A new molecular phylogenetic tree was constructed by the comparison between BcMF20 and these 12 C2H2-type zinc-finger TFs with NJ method. By analyzing this phylogenetic tree, the evolution of BcMF20 was discussed. Then, antisense RNA technology was applied in the transgenesis of Arabidopsis thaliana to get the deletion mutants of BcMF20, so that its function during the pollen development can be identified. The results showed: BcMF20 are in the same clade with three genes from Arabidopsis. The inhibition of BcMF20 expression led to smaller amounts of and lower rate in germination of pollen and lower rate in fruit setting in certain transgenetic plants. This also led to the complete collapse of pollen grains. By SEM and TEM, pollen morphology and anther development processes were observed. In the middle uninucleate microspore stage, a relatively thin or even no primexine was formed in microspores. This may result in the malformation of the pollen wall and finally cause the deformity of pollens. Above all, it can be indicated that BcMF20 may act as a part of regulation mechanisms of TAZ1 and MS1. Together they play a role in a genetic pathway in the tapetum to act on proliferation of tapetal cells and keep the normal development of pollens.

60 APPLIED LIFE SCIENCES↗

Effect of Acyl Activating Enzyme (AAE) 3 on the growth and development of Medicago truncatula

The Acyl-Activating Enzyme (AAE) 3 gene encodes an oxalyl-CoA synthetase that catalyzes the conversion of oxalate to oxalyl-CoA in a CoA and ATP-dependent manner. Although the biochemical activity of AAE3 has been established, its biological role in plant growth and development remains unclear. To advance our understanding of the role of AAE3 in plant growth and development, we report here the characterization of two Medicago truncatula AAE3 (Mtaae3) mutants. Characterization of a Mtaae3 RNAi mutant revealed an accumulation of calcium oxalate crystals and increased seed permeability. These phenotypes were also exhibited in the Arabidopsis aae3 (Ataae3) mutants. Unlike the Ataae3 mutants, the Mtaae3 RNAi mutant did not show a reduction in vegetative growth, decreased seed germination, or increased seed calcium concentration. In an effort to clarify these phenotypic differences, a Mtaae3 Tnt1 mutant was identified and characterized. This Mtaae3 Tnt1 mutant displayed reduced vegetative growth, decreased seed germination, and increased seed calcium concentration as well as an accumulation of calcium oxalate crystals and increased seed permeability as found in Ataae3. Overall, the results presented here show the importance of AAE3 in the growth and development of plants. In addition, this study highlights the ability to separate specific growth and development phenotypes based on the level of AAE3 gene expression.

60 APPLIED LIFE SCIENCES↗

An experimental and kinetic modeling study of the oxidation of hexane isomers: Developing consistent reaction rate rules for alkanes

Alkanes are key components in gasoline, jet and diesel fuels and considerably influence the combustion behavior of these fuels because of their wide range of reactivity. An improved understanding of their combustion behavior and the development of chemical kinetic models that can accurately simulate their combustion behavior are important for the development of next-generation internal-combustion and gas-turbine engines. The current work provides here improved insight into oxidation mechanisms of a representative family of hydrocarbon fuels, specifically the hexane isomers: n-hexane, 2-methylpentane, 3-methylpentane, 2,2-dimethylbutane and 2,3-dimethylbutane. These isomers provide carbon “skeletons” ranging from straight-chained to highly-branched and provide a framework for the subsequent development of kinetic mechanisms for larger alkanes. New ignition delay times for the four branched hexane isomers were measured in a high-pressure shock tube and in a rapid compression machine, both at stoichiometric conditions (φ = 1), p = 15 bar and X O2 = 21% over temperatures from 600 to1300 K. These data were combined with previously published measurements under the same conditions for the remaining n-hexane isomer to provide a complete body of experimental data for kinetic modeling analysis. In addition, very recent experimental measurements of individual intermediate chemical species concentrations from all five hexane isomers in a jet-stirred reactor are also included and provide another family of data for further assessment of hexane isomer reactivity. Different reactivities were observed for each hexane isomer in each experimental facility, resulting from differences in their molecular structures. Consistent reaction rate rules have been applied to develop a combined detailed chemical kinetic model for all five hexane isomers. Kinetic model validation studies are reported to show that the current model reproduces well the ignition delay times of all five alkane isomers, as well as their variations in reactivity over a wide range of temperatures and other operating conditions. Equally important, these results show that it is not necessary to have a separate, different kinetic model for each isomer of a family of alkane fuels and that a single, coherent, integrated set of reaction rate classes and rules is sufficient to accurately describe combustion rates of combustion of straight-chain n-alkanes and branched-chain alkane fuels. This suggests strongly that a single set of reaction classes and rate rules should be sufficient to describe combustion kinetics of alkane fuels of any size and degree of branching.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Influence of PRE-emergence herbicides on soybean development, root nodulation and symbiotic nitrogen fixation

We report widespread occurrence of herbicide-resistant weeds has further limited effective POST-emergence herbicide options in soybean (Glycine max (L.) Merr.) leading to an increased adoption of PRE-emergence herbicides. The objective of this study was to investigate the influence of 11 commonly used PRE herbicides on soybean development, root nodulation, and symbiotic N fixation. Soybean plants were grown under greenhouse conditions in pots (10 L; 4 plants per pot) filled with silt loam soil and treated one day after planting with a labeled field rate of imazethapyr, chlorimuron-ethyl, cloransulam-methyl, metribuzin, sulfentrazone, flumioxazin, saflufenacil, acetochlor, S-metolachlor, dimethenamid-P, pyroxasulfone and no herbicide (nontreated control). Sulfentrazone reduced soybean canopy at the VC growth stage but no canopy reduction was observed at the V2 growth stage from any of the herbicide treatments. At the R2 growth stage, herbicides had no effect on soybean development (root and shoot biomass), root nodulation (# nodule per plant, nodule diameter, and nodule biomass) and symbiotic N fixation (acetylene reduction assay and 15 N natural abundance). According to our findings, although PRE herbicides may slightly affect early-season soybean development, the impacts on plant growth, root nodulation, and symbiotic N fixation were negligible. Thus, when sprayed according to the label, the benefits of PRE herbicides for weed control likely outweigh any potential concern regarding soybean development, root nodulation, and N fixation.

54 ENVIRONMENTAL SCIENCES↗

Urban gray vs. urban green vs. soil protection — Development of a systemic solution to soil sealing management on the example of Germany

Managing urban soil sealing is a difficult venture due to its spatial heterogeneity and embedding in a socio-ecological system. A systemic solution is needed to tackle its spatial, ecological and social sub-systems. This study develops a guideline for urban actors to find a systemic solution to soil sealing management based on two case studies in Germany: Munich and Leipzig. Legal-planning, informal-planning, economic-fiscal, co-operative and informational responses were evaluated by indicators to proof which strategy considers the spatial complexity of urban soil sealing (systemic spatial efficiency) and, while considering spatial complexity, to assess what the key management areas for action are to reduce the ecological impacts by urban soil sealing (ecological impact efficiency) and to support an efficient implementation by urban actors (social implementation efficiency). Results suggest framing the systemic solution to soil sealing management through a cross-scale, legal-planning development strategy embedded in higher European policies. Within the socio-ecological system, the key management area for action should focus on the protection of green infrastructure being of high value for actors from the European to local scales. Further efforts are necessary to establish a systemic monitoring concept to optimize socio-ecological benefits and avoid trade-offs such as between urban infill development and urban green protection. This place-based study can be regarded as a stepping stone on how to develop systemic strategies by considering different spatial sub-targets and socio-ecological systems. - Highlights: • Urban soil sealing management is spatially complex. • The legal-planning strategy supports a systemic sealing management. • Urban green infrastructure protection should be in the management focus. • Soil protection requires policies from higher levels of government. • A systemic urban soil sealing monitoring concept is needed.

54 ENVIRONMENTAL SCIENCES↗

Interpreting best available technologies more flexibly: A policy perspective for municipal wastewater management in India and other developing countries

Inadequate sanitation is amongst the causes of escalating pollution problems in developing countries, as municipal wastewater treatment systems remove only a fraction of pollutants that could be removed with best available technologies (BAT). Although BAT is a proven instrument of environmental policies, its potential for municipalities remains largely unused in developing countries. In order to ease its implementation, the paper developed a simplified assessment approach towards identifying an approximating of BAT in terms of a “flexible BAT” (FlexiBAT), which is based on the identification of national reference plants assessed with respect to pollutant removal (environmental impact, health impact), costs (economic viability, affordability) and social acceptability. The concept was tested for 58 case studies in India, where none of the technologies passed all tests for FlexiBAT. Therefore, there is a need to improve or develop better and more innovative technologies. Amongst the most promising ones, membrane bioreactors provided good physical water quality, but costs were high, while for moving bed biofilm reactors costs were low, but water quality was insufficient. Conventional onsite systems require separate consideration. In order to ease the identification of FlexiBAT, a national environmental information system with data from the regular monitoring of existing plants would be needed.

54 ENVIRONMENTAL SCIENCES↗

Development, calibration, and validation of a novel gray-box energy model for residential split air conditioners

Energy models for vapor compression refrigeration in residential air conditioners have been developed through white-box, gray-box, and black-box methods in decades. However, existing white-box and gray-box models require complicated equations with detailed geometries while black-box models require substantial experimental data. Further, this paper aims to develop and validate a simple gray-box steady-state energy model without the need for detailed geometries, which can accurately predict the cooling capacity and electrical power input based on outdoor and indoor air conditions, and supply airflow rates. First three state variables, including the evaporation and condensation temperatures, and refrigerant mass flowrate, are applied to develop the energy model and are solved by three physical equations, including the energy conservations at the evaporator and condenser, and the refrigerant volume-mass flow correlation of the compressor. Secondly, seven performance property functions related to three state variables and three physical equations are identified and calibrated by simple temperature and power measurements. Finally, field experiments are conducted on a residential air conditioner to calibrate these performance property functions and validate the developed model. The validated results reveal the model can accurately predict the cooling capacity and electrical power input, with the normalized root mean square errors of 2.3% and 0.87% respectively.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Developing energy flexibility in clusters of buildings: A critical analysis of barriers from planning to operation

This paper examines building energy flexibility at an aggregated level and addresses the main barriers and research gaps for the development of this resource across three design and development phases: market and policy, early planning and design, and operation. We review methodologies and tools and discuss barriers, challenges, and opportunities, incorporating policy, economic, technical, professional, and social perspectives. Although various legal and regulatory frameworks exist to foster the development of energy flexibility for small buildings, financing mechanisms are limited with a significant number of perceived risks undermining private investment. For the early planning and design phase, planners and designers lack appropriate tools and face interoperability challenges, which often results in insufficient consideration of demand response programs. The review of the operational phase highlighted the socio-technical challenges related to both the complexity of deployment and communication, as well as privacy and acceptability issues. Finally, the paper proposes a number of targeted research directions to address challenges and promote greater energy flexibility deployments, including capturing building demand side dynamics, improving baseline estimations and developing seamless connectivity between buildings and districts.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Defining pathways to healthy sustainable urban development

Goals and pathways to achieve sustainable urban development have multiple interlinkages with human health and wellbeing. However, these interlinkages have not been examined in depth in recent discussions on urban sustainability and global urban science. This paper fills that gap by elaborating in detail the multiple links between urban sustainability and human health and by mapping research gaps at the interface of health and urban sustainability sciences. As researchers from a broad range of disciplines, we aimed to: 1) define the process of urbanization, highlighting distinctions from related concepts to support improved conceptual rigour in health research; 2) review the evidence linking health with urbanization, urbanicity, and cities and identify cross-cutting issues; and 3) highlight new research approaches needed to study complex urban systems and their links with health. This novel, comprehensive knowledge synthesis addresses issue of interest across multiple disciplines. Our review of concepts of urban development should be of particular value to researchers and practitioners in the health sciences, while our review of the links between urban environments and health should be of particular interest to those outside of public health. We identify specific actions to promote health through sustainable urban development that leaves no one behind, including: integrated planning; evidence-informed policy-making; and monitoring the implementation of policies. We also highlight the critical role of effective governance and equity-driven planning in progress towards sustainable, healthy, and just urban development.

54 ENVIRONMENTAL SCIENCES↗

Temporal nitrogen dynamics in intensively managed loblolly pine early stand development

Forest production is strongly dependent on nutrient uptake; however, sustainable management of intensively managed plantations requires an improved understanding of this relationship when fertilization occurs frequently across short rotations. Here, we studied temporal nitrogen (N) concentration ([N]) and content (Nc) dynamics under different silvicultural practices (herbicide, fertilization, and planting density) throughout early loblolly pine (Pinus taeda) stand development (5 years). We describe relationships of [N] and Nc of different stand components (foliage, branches, stem, roots, and competing vegetation) with carbon and biomass. Our results demonstrate that [N] of perennial loblolly tissues do not respond to silvicultural practices and progressively decrease through development. While foliar [N] was most responsive to resource availability, it was not consistent across time. Controlling competing vegetation was crucial to promote the use of site resources by the crop tree and increased loblolly Nc by >500%. However, increased N uptake and expedited growth is dependent upon fertilization early in stand development. At age 5, herbicide plus reduced and full fertilization rates exhibited similar aboveground Nc, which was 32% higher than with herbicide only. Increasing planting density resulted in increased above- and belowground loblolly Nc; however, increases in Nc were not proportional with increases in planting density. Net primary productivity and N uptake were linearly related, but age/development strongly controlled N use efficiency. Our study helps to understand complex relationships between N, biomass, and silvicultural practices during early stand development and demonstrates that temporal evaluation of nutrient dynamics is crucial to better understand loblolly pine growth, carbon sequestration potential, and to inform sustainable silvicultural practices across short rotations.

54 ENVIRONMENTAL SCIENCES↗