Engineering Papers⌕ Search

SEARCH · Engineering Papers

Results for “Biomass characterization”

Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 487 records · Page 27

Foliar elemental distribution in Blepharidium guatemalense assessed by synchrotron X-ray fluorescence spectroscopy and colorimetric optical microscopy

The hyperaccumulation phenomenon holds significant potential for Ni agromining in ultramafic areas; however, data on tropical hyperaccumulator species, particularly regarding metal distribution and tolerance mechanisms, remain scarce. Here, this study characterized the foliar elemental distribution of Blepharidium guatemalense, analyzed its metal localization at both tissue and cellular levels, and quantified its Ni accumulation when grown in Brazilian ultramafic soil. Plants were grown in pots for 90 days, and Ni concentrations in the aerial biomass and bio-ore were determined. Synchrotron-based micro-X-ray fluorescence (S-µXRF) was used to assess metal distribution in tissues, supplemented by microscopic examination with dimethylglyoxime (DMG) to visualize Ni accumulation in specific cell types. B. guatemalense reached 19,000 mg kg −1 Ni in leaves. Processing leaf biomass produced ash containing 24 wt% Ni, indicating promising metal-recovery potential for the species. S-µXRF revealed that Ni was predominantly localized in the central and secondary veins of the leaves, suggesting efficient vascular transport. Microscopic analysis using the colorimetric reagent DMG further showed that Ni accumulation occurs near the epidermis and phloem, suggesting that B. guatemalense employs specific physiological mechanisms for Ni translocation, potentially supporting both defense and growth functions. These results establish B. guatemalense as a promising candidate for large-scale Ni agromining in tropical regions.

36 MATERIALS SCIENCE↗

Chemical composition of the atmospheric aerosol in the troposphere over the Hudson Bay lowlands and Quebec-Labrador regions of Canada

Atmospheric aerosols were collected in the boundary layer and free troposphere over continental and coastal subarctic regions of Canada during the July - August 1990 joint U.S.-Canadian Arctic Boundary Layer Expedition (ABLE) 3B/Northern Wetlands Study (NOWES). The samples were analyzed for the following water soluble species: sulfate, nitrate, ammonium, potassium, sodium, chloride, oxalate, methylsulfonate, and total amine nitrogen. Ammonium and sulfate were the major water soluble components of these aerosols. The nearly neutral (overall) chemical composition of summertime aerosol particles contrasts their strongly acidic wintertime composition. Aerosol samples were separated into several air mass categories and characterized in terms of chemical composition, associated mixing ratios of gaseous compounds, and meteorological parameters. The fundamental category represented particles associated with 'background' air masses. The summertime atmospheric aerosols in background air over the North American subarctic and Arctic regions were characterized by relatively small and spatially uniform mixing ratios of the measured species. These aerosol particles were aged to the extent that they had lost their primary source signature. The chemical profile of the background air aerosols was frequently modified by additions from biomass fire plumes, aged tropical marine air, and intrusions of upper tropospheric/lower stratospheric air. Aerosols in boundary layer background air over the boreal forest region of Quebec-Labrador had significantly larger mixing ratios of ammonium and sulfate relative to the Hudson Bay region. This may reflect infiltration of anthropogenic pollution or be due to natural emissions from this region.

Gorzelska, K.↗

Competitive Exclusion and Metabolic Dependency among Microorganisms Structure the Cellulose Economy of an Agricultural Soil

Microorganisms that degrade cellulose utilize extracellular reactions that yield free by-products which can promote interactions with noncellulolytic organisms. We hypothesized that these interactions determine the ecological and physiological traits governing the fate of cellulosic carbon (C) in soil. We performed comparative genomics with genome bins from a shotgun metagenomic-stable isotope probing experiment to characterize the attributes of cellulolytic and noncellulolytic taxa accessing 13 C from cellulose. We hypothesized that cellulolytic taxa would exhibit competitive traits that limit access, while noncellulolytic taxa would display greater metabolic dependency, such as signatures of adaptive gene loss. We tested our hypotheses by evaluating genomic traits indicative of competitive exclusion or metabolic dependency, such as antibiotic production, growth rate, surface attachment, biomass degrading potential, and auxotrophy. The most 13 C-enriched taxa were cellulolytic Cellvibrio (Gammaproteobacteria) and Chaetomium (Ascomycota), which exhibited a strategy of self-sufficiency (prototrophy), rapid growth, and competitive exclusion via antibiotic production. Auxotrophy was more prevalent in cellulolytic Actinobacteria than in cellulolytic Proteobacteria, demonstrating differences in dependency among cellulose degraders. Noncellulolytic taxa that accessed 13 C from cellulose (Planctomycetales, Verrucomicrobia, and Vampirovibrionales) were also more dependent, as indicated by patterns of auxotrophy and 13 C labeling (i.e., partial labeling or labeling at later stages). Major 13 C-labeled cellulolytic microbes (e.g., Sorangium, Actinomycetales, Rhizobiales, and Caulobacteraceae) possessed adaptations for surface colonization (e.g., gliding motility, hyphae, attachment structures) signifying the importance of surface ecology in decomposing particulate organic matter. Our results demonstrated that access to cellulosic C was accompanied by ecological trade-offs characterized by differing degrees of metabolic dependency and competitive exclusion.

59 BASIC BIOLOGICAL SCIENCES↗

A distinct class of ferredoxin:NADP + oxidoreductase enzymes driving thermophilic ethanol production

Biofuel production from lignocellulosic biomass offers a transformative solution to reduce global fossil fuel dependency. Certain thermophilic anaerobes, including Clostridium thermocellum, show promise for renewable ethanol production due to their ability to break down plant material at high temperatures. However, achieving commercially viable ethanol yields has proven challenging despite extensive engineering efforts. Here, we characterized 27 ferredoxin:NADP + oxidoreductase (Fnor) enzymes for their enzyme activity, nicotinamide cofactor specificity, thermotolerance, and functional expression in C. thermocellum . We identified a subset of 10 of these enzymes as a novel class of Fnor enzymes suited for metabolic pathways aimed at high-titer ethanol production. When expressed in engineered C. thermocellum , these enzymes increased ethanol production up to 2.2-fold. These findings establish a novel ethanol pathway and provide insights into physiological roles and biotechnological applications of this new class of Fnor enzymes.

09 BIOMASS FUELS↗

AIS spectra of desert shrub canopies

Airborne Imaging Spectrometer (AIS) data were collected 30 August 1985 from a desert shrub community in central Oregon. Spectra from artificial targets placed on the test site and from bare soil, big sagebrush (Artemesia tridentata wyomingensis), silver sagebrush (Artemesia cana bolander), and exposed volcanic rocks were studied. Spectral data from grating position 3 (tree mode) were selected from 25 ground positions for analysis by Principal Factor Analysis (PFA). In this grating position, as many as six factors were identified as significant in contributing to spectral structure. Channels 74 through 84 (tree mode) best characterized between-class differences. Other channels were identified as nondiscriminating and as associated with such errors as excessive atmospheric absorption and grating positin changes. The test site was relatively simple with the two species (A. tridentata and A. cana) representing nearly 95% of biomass and with only two mineral backgrounds, a montmorillonitic soil and volcanic rocks. If, as in this study, six factors of spectral structure can be extracted from a single grating position from data acquired over a simple vegetation community, then AIS data must be considered rich in information-gathering potential.

Murray, R.↗

Aerosol Optical Properties at the Lulin Atmospheric Background Station in Taiwan and the Influences of Long-Range Transport of Air Pollutants

The Lulin Atmospheric Background Station (LABS, 23.47 deg. N 120.87 deg. E, 2862 m ASL) in Central Taiwan was constructed in 2006 and is the only high-altitude background station in the western Pacific region for studying the influence of continental outflow. In this study, extensive optical properties of aerosols, including the aerosol light scattering coefficient [Sigma(sub s)] and light absorption coefficient [Sigma(sub a)], were collected from 2013 to 2014. The intensive optical properties, including mass scattering efficiency [Sigma(sub s)], mass absorption efficiency [Sigma(sub a)] single scattering albedo (Omega), scattering Angstrom exponent (A), and backscattering fraction (b), were determined and investigated, and the distinct seasonal cycle was observed. The value of [Alpha(sub a)] began to increase in January and reached a maximum in April; the mean in spring was 5.89 m(exp. 2) g(exp. -1) with a standard deviation (SD) of 4.54 m(exp. 2) g(exp. -1) and a 4.48 m(exp. 2) g(exp. -1) interquartile range (IQR: 2.95-7.43 m(exp. 2) g(exp. -1). The trend was similar in [Sigma(sub a)], with a maximum in March and a monthly mean of 0.84 m(exp. 2) g(exp. -1). The peak values of Omega (Mean = 0.92, SD = 0.03, IQR: 0.90 - 0.93) and A (Mean = 2.22, SD = 0.61, IQR: 2.12 = 2.47) occurred in autumn. These annual patterns of optical properties were associated with different long-range transport patterns of air pollutants such as biomass burning (BB) aerosol in spring and potential anthropogenic emissions in autumn. The optical measurements performed at LABS during spring in 2013 were compared with those simultaneously performed at the Doi Ang Kang Meteorology Station, Chiang Mai Province, Thailand (DAK, 19.93 deg. N, 99.05 deg. E, 1536 m a.s.l.), which is located in the Southeast Asia BB source region. Furthermore, the relationships among [Sigma(sub s)], [Sigma(sub a)], and (b) were used to characterize the potential aerosol types transported to LABS during different seasons, and the data were inspected according to the HYSPLIT 5-day backward trajectories, which differentiate between different regions of air mass origin.

aerosol optical properties↗

A Catabolic Powerhouse for Biorefineries: Characterization and Engineering Erwinia spp. Strain LJJL01 to Produce Bioproducts

Nonmodel microbial hosts can efficiently biotransform unconventional organic feedstocks into advanced bioproducts, leveraging their superior metabolic capabilities and resilience to process-relevant physicochemical conditions. In this study, we domesticated Erwinia spp. strain. LJJL01 (Er LJJL01) as a potent microbial chassis to advance the emerging biorefinery strategy for the valorization of lignin-rich biomass and plastic, thereby enabling the circular economy. The strain exhibits remarkable chemical tolerance and can biofunnel various substrates, including sugars, acids, polyols, and aromatics, in minimal salt media to produce native fine chemicals such as acetoin, 2,3-butanediol, and lactic acid. As a proof of concept, engineering this strain with advanced genetic tools enables the production of tailored high-value chemicals, such as cis,cis-muconate from plastic-derived terephthalate and polyhydroxybutyrate from lignocellulosic hydrolysate. We established Er LJJL01 as a potent microbial chassis for the green synthesis of bioproducts from unconventional organic feedstocks.

09 BIOMASS FUELS↗

Characterization of terrestrial ecosystems for biogeochemical studies using remote sensing

Work is in progress to estimate leaf area index (LAI) of temperate closed canopy coniferous forests using transects in Oregon and California. This variable will be measured using remote sensing techniques including correlations of ground dimensional analysis with linear waveband combinations. LAI will be related to important biological variables such as net primary productivity, biomass, and biogenic gas emission fluxes. The spatial variation in LAI, when coupled with species composition, will be used in part to describe the spatial variation and temporal dynamics of biogeochemical cycling.

Peterson, D. L.↗

Development of the high-productivity marine microalga, Picochlorum renovo , as a photosynthetic protein secretion platform

Microalgae are remarkable in their ability to convert CO 2 and sunlight into renewable biomass and bioproducts. In this work, we have established a novel photosynthetic protein production platform via computational prediction and genetic engineering using native microalgal protein secretory signal peptides to achieve functional secretion of the fluorescent protein mCherry. Importantly, this work was conducted in the recently characterized alga Picochlorum renovo, an alga of industrial interest due to its rapid growth rate, tolerance to both high temperature and salinity, and genetically tractable nuclear and chloroplast genomes. Genomic queries allowed the identification of native secretory signal peptides, which were N-terminally fused to mCherry allowing for secretion into the culture supernatant. Further characterization revealed no impact on fitness, a production rate of 0.19 mg/L/day, and titer of 0.37 mg/L of transgenic mCherry protein in culture supernatant. These findings lay the foundation for applied genetic engineering approaches that could enable CO 2 -sequestering, sustainable photoproduction of industrially relevant enzymes at low cost.

59 BASIC BIOLOGICAL SCIENCES↗

A single amino acid change led to structural and functional differentiation of PvHd1 to control flowering in switchgrass

Abstract Switchgrass, a forage and bioenergy crop, occurs as two main ecotypes with different but overlapping ranges of adaptation. The two ecotypes differ in a range of characteristics, including flowering time. Flowering time determines the duration of vegetative development and therefore biomass accumulation, a key trait in bioenergy crops. No causal variants for flowering time differences between switchgrass ecotypes have, as yet, been identified. In this study, we mapped a robust flowering time quantitative trait locus (QTL) on chromosome 4K in a biparental F2 population and characterized the flowering-associated transcription factor gene PvHd1, an ortholog of CONSTANS in Arabidopsis and Heading date 1 in rice, as the underlying causal gene. Protein modeling predicted that a serine to glycine substitution at position 35 (p.S35G) in B-Box domain 1 greatly altered the global structure of the PvHd1 protein. The predicted variation in protein compactness was supported in vitro by a 4 °C shift in denaturation temperature. Overexpressing the PvHd1-p.35S allele in a late-flowering CONSTANS-null Arabidopsis mutant rescued earlier flowering, whereas PvHd1-p.35G had a reduced ability to promote flowering, demonstrating that the structural variation led to functional divergence. Our findings provide us with a tool to manipulate the timing of floral transition in switchgrass cultivars and, potentially, expand their cultivation range.

59 BASIC BIOLOGICAL SCIENCES↗

Deinococcus radiodurans UWO298 Dependence on Background Radiation for Optimal Growth

Ionizing radiation is a major environmental variable for cells on Earth, and so organisms have adapted to either prevent or to repair damages caused by it, primarily from the appearance and accumulation of reactive oxygen species (ROS). In this study, we measured the differential gene expression in Deinococcus radiodurans UWO298 cultures deprived of background ionizing radiation (IR) while growing 605 m underground at the Waste Isolation Pilot Plant (WIPP), reducing the dose rate from 72.1 to 0.9 nGy h –1 from control to treatment, respectively. This reduction in IR dose rate delayed the entry into the exponential phase of the IR-shielded cultures, resulting in a lower biomass accumulation for the duration of the experiment. The RNASeq-based transcriptome analysis showed the differential expression of 0.2 and 2.7% of the D. radiodurans genome after 24 and 34 h of growth in liquid culture, respectively. Gene expression regulation after 34 h was characterized by the downregulation of genes involved in folding newly synthesized and denatured/misfolded proteins, in the assimilation of nitrogen for amino acid synthesis and in the control of copper transport and homeostasis to prevent oxidative stress. We also observed the upregulation of genes coding for proteins with transport and cell wall assembly roles. These results show that D. radiodurans is sensitive to the absence of background levels of ionizing radiation and suggest that its transcriptional response is insufficient to maintain optimal growth.

59 BASIC BIOLOGICAL SCIENCES↗

Wood combustion nanoparticles emitted by conventional and advanced technology cordwood boilers, and their interactions in vitro with human lung epithelial monolayers

Biomass-burning boilers and stoves are widely used in many parts of the world, producing combustion emissions linked with health risks. Here, combustion emission nanoparticles (NPs) were collected from four representative wood burning boilers using oak cordwood at specific times in the burn cycle. The morphology and composition of the NPs was characterized using transmission electron microscopy and energy dispersive X-ray analysis. To determine the degree of NP cytotoxicity with human lung tissue, the combustion NPs were introduced to incubated lung bronchial epithelial monolayers (NCI-H292) in vitro at doses of 0.1×10 -6 and 3.0×10 -6 kg/L for 2 and 4 h. Histochemical analysis showed that cell death increased by a factor of 3.5 for both doses after 4 h when compared to the control. Ultrapure NPs prepared by wet chemical methods were also introduced to the epithelial lung cells for similar doses and exposure times and the cultures exhibited significantly reduced mortality. Electron microscopy was used to study the mechanism of cell mortality for the synthesized and combustion-based NPs by examining how the NP byproducts interacted with individual cell organelles. It was found that cell survival was strongly correlated with the absence of contaminants (salts, heavy metals, poly aromatic hydrocarbons) associated with the NPs entering the cells. Synthesized NPs consisting of pure carbon were relatively well tolerated and could be excreted without damaging the cell ultrastructure. Thus, careful removal of extraneous contaminants by controlling the burn cycle with a catalyst is essential to minimize the health and environmental effects of wood biofuel combustion. In better words, optimized advanced technology wood-burning boilers and stoves can provide a CO 2 -neutral energy source and significantly contribute to a future where fossil fuels have a reduced role.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Rapid in situ nutrient element distribution in plants and soils using laser-induced breakdown spectroscopy ($\mathrm{LIBS}$)

The aim of this study is to develop and test the applicability of a rapid in situ plant chemistry profiling technique to determine elemental composition of small-volume plant and soil samples obtained from a woody bioenergy crop species, Populus trichocarpa. Expanding the research tools available to characterize the nutrient element correlations among plant tissue types and soil depths is a critical need in the path of understanding productivity and adaptation of plants to variations in external abiotic and biotic factors and developing sustainable perennial bioenergy crops that are co-optimized for biomass valorization aboveground and carbon sequestration belowground. Several plant root, stem, and soil samples were tested using laser-induced breakdown spectroscopy (LIBS) to evaluate the presence and distribution of nutrient elements. Samples were tested as collected and after being dried and cross sectioned to evaluate the effectiveness of using LIBS for in situ analysis on plant samples. The collected LIBS spectra show the elemental peaks were the same in both the as collected and prepared samples for roots and stems. Qualitative amounts of elements such as H, C, N, O, Li, Na, Mg, K, Ca, Fe, Al, and Si were able to be identified rapidly in raw samples. Here in this report we demonstrate suitability of LIBS in obtaining rapid, in situ, elemental distribution in plant and soil samples, utilizing only small sample volumes and minimal sample preparation. This demonstration opens up a new rapid phenotyping avenue necessary to fill the asymmetrical knowledge gaps in belowground performance of plant systems.

59 BASIC BIOLOGICAL SCIENCES↗

Regenerative Life Support Systems (RLSS) test bed performance - Characterization of plant performance in a controlled atmosphere

By growing higher plants for food, lunar and Martian manned habitats will not only reduce resupply requirements but obtain CO2 removal and both oxygen-production and water-reclamation requirements. Plants have been grown in the RLSS at NASA-Johnson in order to quantitatively evaluate plant CO2 accumulation, O2 generation, evapotranspiration, trace-contaminant generation, and biomass productivity. Attention is presently given to test conditions and anomalies in these RLSS trials; areas where performance must be improved have been identified.

Edeen, Marybeth↗

Insights into Mechanisms of Screw Feeder Plugging by Heated Pine Forestry Residues Using in-situ and Correlative Microscopy

Increasing concerns about global carbon emissions due to fossil fuel use has created interest in renewable fuel alternatives, such as lignocellulosic biomass. With an estimated global biomass production probability ten-fold higher than projected need, functionalizing and scaling biomass conversion processes, like pyrolysis, is a promising component of a diverse sustainable energy program (Kan, Strezov, & Evans, 2016). Pyrolysis reactors employed in biomass conversion often encounter the known problem of screw feeder plugging, halting work and reducing efficiency of biorefineries. Although the screw feeder is not heated directly, conductive heat transfer from the fluidized bed pyrolysis reactor creates a temperature gradient, where particles experience temperatures between 22°C and 500°C from the time they enter the feeder to being emptied into the reactor (Digman, Joo, & Kim, 2009; Nieminen & Folke, 1982). The impact of unintentional heating on this highly variable feedstock may result in particle morphology modification, surface texture transformation, and the evolution of viscous compounds and may contribute to screw feeder plugging (Haas, Nimlos, & Donohoe, 2009; Ingram et al., 2008). We hypothesized that particles of various anatomical fractions (including needles, branches, bark, cambium, and whitewood) will experience dramatic changes during this heating regime, which will be characterized by decreased particle size, increased surface roughness, degradation of key structural macromolecules, and generation of viscous compounds that would result in increased particle friction and cohesion, contributing to screw feeder plugging.

biofuels↗

Let’s Stick Together: Interplay of Moisture, Particle Size, and Anatomical Fraction in the Flowability of Corn Stover Crumbles

Handling of agricultural waste biomass feedstocks (such as corn stover) is a persistent challenge in the production of biofuels and chemicals at integrated biorefineries. Inconsistent flowability and high feedstock variability creates equipment downtime and processing challenges. This work investigates the influence of anatomical fractionation and moisture content on the bulk solids handling behavior of corn stover crumbles. A combination of rotational shear testing, tribological measurements, and surface and sorption characterization techniques (inverse gas chromatography (IGC), dynamic vapor sorption (DVS), and electron microscopy) were used to evaluate the contributions of particle composition, size, and surface properties to flow behavior. Shear and frictional testing revealed that moisture content has the greatest effect on flowability, significantly increasing unconfined yield strength (ƒ c ) and reducing the flow function coefficient (F F C), regardless of anatomical fraction. While differences in surface energy were observed between fractions, particularly in polar contributions (γ AB ), these did not correspond to meaningful differences in flow behavior. Coarser 6 mm particles exhibited poorer flow performance than 4 mm crumbles, likely due to broader particle size distributions and increased particle interlocking. Taken together, these results suggest that anatomical fractionation provides minimal benefit from a bulk handling perspective. Moisture content and particle size heterogeneity dominate handling behavior, reinforcing the utility of unfractionated corn stover in processing environments. These insights can inform the design of more efficient and reliable feedstock processing systems for agricultural biomass in integrated biorefineries.

09 BIOMASS FUELS↗

In-depth understanding of molecular mechanisms of aldehyde toxicity to engineer robust Saccharomyces cerevisiae

Aldehydes are ubiquitous electrophilic compounds that ferment microorganisms including Saccharomyces cerevisiae encounter during the fermentation processes to produce food, fuels, chemicals, and pharmaceuticals. Aldehydes pose severe toxicity to the growth and metabolism of the S. cerevisiae through a variety of toxic molecular mechanisms, predominantly via damaging macromolecules and hampering the production of targeted compounds. Compounds with aldehyde functional groups are far more toxic to S. cerevisiae than all other functional classes, and toxic potency depends on physicochemical characteristics of aldehydes. The yeast synthetic biology community established a design–build–test–learn framework to develop S. cerevisiae cell factories to valorize the sustainable and renewable biomass, including the lignin-derived substrates. However, thermochemically pretreated biomass-derived substrate streams contain diverse aldehydes (e.g., glycolaldehyde and furfural), and biological conversions routes of lignocellulosic compounds consist of toxic aldehyde intermediates (e.g., formaldehyde and methylglyoxal), and some of the high-value targeted products have aldehyde functional group (e.g., vanillin and benzaldehyde). Numerous studies comprehensively characterized both single and additive effects of aldehyde toxicity via systems biology investigations, and novel molecular approaches have been discovered to overcome the aldehyde toxicity. Based on those novel approaches, researchers successfully developed synthetic yeast cell factories to convert lignocellulosic substrates to valuable products, including aldehyde compounds. Here, we highlight the salient relationship of physicochemical charac- teristics and molecular toxicity of aldehydes, the molecular detoxification and macromolecules protection mechanisms of aldehydes, and the advances of engineering robust S. cerevisiae against complex mixtures of aldehyde inhibitors.

59 BASIC BIOLOGICAL SCIENCES↗

Engineered enzymatic cascade converts diols to amino alcohols

Aliphatic amino alcohols such as 6-amino-1-hexanol are potential platform chemicals for a variety of advanced materials, but applications are currently limited by reagent costs. Aliphatic amino alcohols can currently be synthesized from biomass-derived diols at elevated temperatures and pressures using Ru-based catalysts that produce a mixture of amino-alcohol, diamine, and cyclic amine products. Replacing chemical amination with an enzymatic cascade would reduce resource needs and enable reactions under milder conditions. In this work, we characterized a two-enzyme cascade that selectively converts C4–C7 diols to the corresponding amino alcohols under aqueous conditions at room temperature and pressure. By engineering the rate-limiting enzyme and optimizing reaction conditions, we increased amino alcohol production nearly 30-fold, achieving a selectivity of 99%. The same enzyme cascade could also be used to convert amino alcohols into cyclic amines through reduction of the corresponding cyclic imine. This engineered cascade provides a green opportunity to sustainably synthesize asymmetric bifunctional platform chemicals.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗