Nonconjugated amylopectin-grafted copolymers with dual fluorescence/low-temperature phosphorescence emission and superior processability.

Nonconjugated fluorescent polymers devoid of large π-π conjugated structures have received considerable attention due to their significant academic importance and broad application potentials in various fields. Herein, we report an effective strategy to fabricate multifunctional fluorescent amylopectin derivatives and reveal their unique aspects of aggregation-induced emission (AIE), cryogenic long-persistent phosphorescence (~6 s) and excellent processabilities characteristics, which are extremely different from traditional luminogens. These amylopectin-graft-poly(n-butyl acrylate-co-1-vinylimidazole) copolymers (Amylopectin-BVs) prepared by the grafting-from method employing RAFT and experienced subsequently with metal-ligand cross-linking. Specifically, clustering-triggered fluorescent emission or cryogenic long-persistent phosphorescence of amylopectin could be achieved by the aggregation of oxygen and nitrogen atoms along with conformation rigidification, which shows great promise in optoelectronic and biological applications.

Photo-crosslinking methacrylated-amylopectin/polyacrylamide hydrogels loading curcumin for applications as degradable, injectable, and antibacterial wound dressings.

The preparation of biodegradable and antibacterial hydrogels has important clinical value. In this work, a novel strategy has been developed to prepare degradable hydrogel dressings without chemical crosslinking agent using methacrylate anhydride (MA)-modified amylopectin (APMA) and polyacrylamide (PAM). After introducing CC bonds, APMA/PAM hydrogels can be formed under light irradiation. This strategy improves the gelling ability of AP and degradation properties of the hydrogel by avoiding the addition of crosslinking agent. The degradation rate of APMA/PAM hydrogel is 74.04 ± 0.69 % within 12 weeks, while that of APMA/PAM hydrogel containing crosslinking agent is only 38.5 ± 0.1 %. The APMA/PAM hydrogel loading curcumin (Cur) (APMA/PAM-Cur) exhibits high antibacterial efficiency of 98.29 ± 0.41 % and 97.18 ± 0.81 % against S. aureus and E. coli, respectively, with light irradiation. Animal experiments show that the APMA/PAM-Cur hydrogel reduces the infiltration of inflammatory factors, increases the density of collagen, and makes the newly formed granulation tissue thicker and tighter. This study not only proves the promising potential of the APMA/PAM-Cur hydrogel as degradable and antibacterial wound dressing for clinical treatment, but also provides a new strategy for developing low-cost, degradable, and antibacterial wound dressings and reducing antibiotic abuse and environmental pollution caused by medical waste.

Impact of starch amylose and amylopectin on the rheological and 3D printing properties of corn starch.

This study evaluated the influence of the amylose and amylopectin on the physicochemical properties and printing performance of corn starch gels. Amylose in starch-based gels enhances their storage modulus and the support performance of printed products by promoting the formation of cross-linked gel structures and crystalline structures. However, the higher amylose content in starch gels makes extrusion difficult, resulting in intermittent extrusion in 3D printing. Despite the increased shear-thinning ability of high-amylose starch, its low water retention capacity leads to water loss and rough printed morphology. Additionally, starch with 72 % amylose content exhibits insufficient adhesive properties for effective layer bonding, negatively impacting structural integrity. While gels with 72 % and 56 % amylose content demonstrate higher viscosity and enhanced mechanical properties, their poor adhesion limits the quality of printed layers. Conversely, waxy starch gel demonstrates continuous extrusion and adhesion but lacks adequate support. The 27 % corn starch gel achieves the highest 3D printing accuracy at 88.12 %, suggesting an optimal amylose-amylopectin ratio for desired ink material performance. These findings enhance our understanding of the relationship between amylose content in starch and 3D printing performance, providing a theoretical basis for the development of starch-based printing products.

Starch Characteristics and Amylopectin Unit and Internal Chain Profiles of Indonesian Rice (Oryza sativa).

Indonesia is arguably a major player in worldwide rice production. Though white rice is the most predominantly cultivated, red, brown, and red rice are also very common. These types of rice are known to have different cooking properties that may be related to differences in their starch properties. Investigating the starch properties, especially the fine structure of their amylopectin, can help understand these differences. This study aims to investigate the starch characteristics of some Indonesian rice varieties by evaluating the starch granule morphology and size, molecular characteristics, amylopectin unit and internal chain profiles, and thermal properties. Starches were extracted from white rice (long grain (IR-64) and short grain (IR-42)), brown rice, red rice, and black rice cultivated in Java Island, Indonesia. IR-42 had the highest amylose content of 39.34% whilst the black rice had the least of 1.73%. The enthalpy of gelatinization and onset temperature of the gelatinization of starch granules were between 3.2 and 16.2 J/g and 60.1 to 73.8 °C, respectively. There were significant differences between the relative molar amounts of the internal chains of the samples. The two white rice and black rice had a significantly higher amount of A-chains, but a lower amount of B-chains and fingerprint B-chains (Bfp) than the brown and red rice. The average chain length (CL), short chain length (SCL), and external chain length (ECL) were significantly longer for the red rice and the black rice in comparison to both the white rice amylopectins. The long chain length (LCL) and internal chain length (ICL) of the sample amylopectins were similar. Rice starches were significantly different in the internal structure but not as much in their amylopectin unit chain profile. These results suggest the differences in their amylopectin clusters and building blocks.

Variation Analysis of Starch Properties in Tartary Buckwheat and Construction of Near-Infrared Models for Rapid Non-Destructive Detection.

Due to the requirements for quality testing and breeding Tartary buckwheat (Fagopyrum tartaricum Gaerth), it is necessary to find a method for the rapid detection of starch content in Tartary buckwheat. To obtain samples with a continuously distributed chemical value, stable Tartary buckwheat recombinant inbred lines were used. After scanning the near-infrared spectra of whole grains, we employed conventional methods to analyze the contents of Tartary buckwheat. The results showed that the contents of total starch, amylose, amylopectin, and resistant starch were 532.1-741.5 mg/g, 176.8-280.2 mg/g, 318.8-497.0 mg/g, and 45.1-105.2 mg/g, respectively. The prediction model for the different starch contents in Tartary buckwheat was established using near-infrared spectroscopy (NIRS) in combination with chemometrics. The Kennard-Stone algorithm was used to split the training set and the test set. Six different methods were used to preprocess the spectra in the wavenumber range of 4000-12,000 cm-1. The Competitive Adaptive Reweighted Sampling algorithm was then used to extract the characteristic spectra, and the prediction model was built using the partial least squares method. Through a comprehensive analysis of each parameter of the model, the best model for the prediction of each nutrient was determined. The correlation coefficient of calibration (Rc) and the correlation coefficient of prediction (Rp) of the best models for total starch and amylose were greater than 0.95, and the Rc and Rp of the best models for amylopectin and resistant starch were also greater than 0.93. The results showed that the NIRS-based prediction model fulfilled the requirement for the rapid determination of Tartary buckwheat starch, thus providing an effective technical approach for the rapid and non-destructive testing of starch content in the food science and agricultural industry.

Potential use of other starch sources in the treatment of glycogen storage disease type Ia - an in vitro study.

BACKGROUND: Glycogen storage disease type Ia (GSD-Ia) is one of the most common hepatic GSD. Its treatment mainly consists of a diet including a high intake of slow-digestion carbohydrates such as raw cornstarch and the restriction of simple sugars. This enables the maintenance of euglycemia and prevents secondary metabolic disorders. Starch is a glucose polymer formed by amylose and amylopectin, which can be obtained from distinct sources. Although uncooked cornstarch has been successfully used in the treatment of GSD-Ia, it can lead to hyperglycemia and weight gain. in vitro andin vivo tests indicated that sweet manioc starch can be potentially used in the treatment of GSD-Ia. RESULTS: The moisture analysis revealed a variation from 10.3 to 12.8% in the sweet manioc starch samples, whereas the moisture content of uncooked cornstarch ranged from 7.3 to 11.1%. Quantifiable sugar was detected in 3/5 samples of sweet manioc starch and 1/3 samples of uncooked cornstarch. Notably, this uncooked cornstarch brand is widely employed in GSD-Ia treatment in Brazil. Products B and E had higher values of amylopectin and undetectable levels of sugars. A clinical trial is warranted to compare samples F and G and determine the impact of sugar trace in the same dietary source of starch. CONCLUSIONS: Collectively, the results demonstrated possible therapeutic alternatives for GSD-Ia in addition to traditional uncooked cornstarch.

Highly stretchable, self-healing, antibacterial, conductive, and amylopectin-enhanced hydrogels with gallium droplets loading as strain sensors.

In this study, we address the challenge of developing highly conductive hydrogels with enhanced stretchability for use in wearable sensors, which are critical for the precise detection of human motion and subtle physiological strains. Our novel approach utilizes amylopectin, a biopolymer, for the uniform integration of liquid metal gallium into the hydrogel matrix. This integration results in a conductive hydrogel characterized by remarkable elasticity (up to 7100 % extensibility) and superior electrical conductance (Gauge Factor = 31.4), coupled with a minimal detection limit of less than 0.1 % and exceptional durability over 5000 cycles. The hydrogel demonstrates significant antibacterial activity, inhibiting microbial growth in moist environments, thus enhancing its applicability in medical settings. Employing a synthesis process that involves ambient condition polymerization of acrylic acid, facilitated by a hydrophobic associative framework, this hydrogel stands out for its rapid gelation and robust mechanical properties. The potential applications of this hydrogel extend beyond wearable sensors, promising advancements in human-computer interaction through technologies like wireless actuation of robotic systems. This study not only introduces a viable material for current wearable technologies but also sets a foundation for future innovations in bio-compatible sensors and interactive devices.

Impact of Rare Sugar D-Allulose on Hardening of Starch Gels during Refrigerated Storage.

The rare sugar D-allulose (Alu), with ca. 10% calories of sucrose (Suc), is a promising alternative sugar that can be used to improve the quality of starch gels in storage. The effects of Alu (compared to Suc) on the hardening and microstructural and molecular order of amylopectin-rich (glutinous rice (GR) and corn amylopectin (CAP)) and amylose-rich (corn (C)) starch gels were investigated. Alu and Suc both suppressed hardening in C gels, while Alu but not Suc was effective in GR and CAP gels. SEM results showed that Alu-containing GR and CAP maintained a relatively large pore size compared to Suc-containing gels. The deconvolution of FTIR spectra revealed that Alu-containing GR and CAP gels had lower ratios of intermolecular hydrogen bonds and higher ratios of loose hydrogen bonds than Suc-containing gels. For amylose-rich C gels, on the other hand, such tendencies were not observed. The influence of Alu on amylopectin-rich gels could be because Alu reduced the ratio of intermolecular hydrogen bonds, which might be involved in amylopectin recrystallization, and increased that of loose hydrogen bonds. The results suggest that Alu is more effective than Suc in inhibiting the hardening of amylopectin-rich starch gels during refrigerated storage.

Correlation Study between Multi-Scale Structure and In Vitro Digestibility of Starch Modified by Temperature Difference.

Physical techniques are widely applied in the food industry due to their positive impact on food quality and the environment. Temperature differences can effectively modify starch, but the resulting changes in starch structure and quality remain unclear. In this study, the corn starch was processed with high temperature, low temperature, and temperature difference (TD), including high temperature before low temperature (H-L) and low temperature before high temperature (L-H). The results showed that high temperature induced the umbilicus to concave inward shape and sharply decreased the amylose content, while low temperature increased the surface micropores and reduced the A-chain. TD reduced the fluorescence intensity and increased the clearness of the growth ring. TD elevated the relative crystallinity (RC), short-range order, A/B1 chains, hydrolysis parameters, and resistant starch (RS), and reduced amylose content, B2/B3 chains, and viscosity. Moreover, the corn starches treated by H-L had lower amylose content and higher RC, 1047/1022, A-chain, and RS than those treated by L-H. Overall, high temperature degraded the amylose and low temperature destroyed the amylopectin. During the TD, H-L can accelerate the starch molecular rearrangement more than the opposite temperature treatment order. These results will help produce novel starches for better food applications.

Effects of Binding between Ca in Hard Water and Phosphorus in Amylopectin on the Qualities of Boiled Rice and Rice Noodle Prepared by Soaking and Boiling in Hard Water.

Recently, global warming has led to an increase in chalky rice grains. This has consequently resulted in the deterioration in quality of rice products. Although we previously reported that hard water, rich in Ca, is useful for the quality improvement of high-temperature-damaged rice grains, the mechanism was not elucidated sufficiently. Therefore, we used various kinds of rice cultivars, from waxy to high-amylose ones, for soaking and boiling in hard water and compared physical and chemical properties of the products. It was shown that the degree of quality improvement, such as final viscosities in pasting property, and textural properties of boiled rice, was more remarkable for high-amylose rice than low-amylose rice. As we found that the phosphorus contents showed positive correlations with amylose and long chains of amylopectin, we estimate that the effects are mainly due to binding of calcium and phosphorus. Because that high-amylose or long-chain-rich amylopectin rice cultivars showed high calcium contents in rice products, these rice cultivars would be very useful to supply calcium through dietary intake via hard water cooking.

Constructing an interaction in plant polyphenol-modified carbon fiber with amylopectin-based waterborne polyurethane sizing agent via hydrogen bonding to improve the interfacial performance of carbon fiber/nylon 6 composites.

The adhesive strength between the sizing agent and carbon fiber (CF) plays a crucial role in improving the interfacial properties of composites, while such a vital aspect has been consistently disregarded. In this study, a hyperbranched waterborne polyurethane (HWPU) sizing agent was synthesized from biogenetically raw materials including gallic acid, l-Lysine diisocyanate and amylopectin. Concurrently, hydrogen-bonded cross-linked network structures were established utilizing a botanical polyphenol tannin as coupling agent to effectively connect CF with HWPU. This meticulous process yielded CF/nylon 6 composites with improved properties and their mechanical characteristics were systematically investigated. The findings showcased a noteworthy boost in flexural strength and interlaminar shear strength (ILSS), showing enhancements of 54.6 % and 61.4 %, respectively, surpassing those of untreated CF. Furthermore, the interfacial shear strength (IFSS) test indicated a remarkable 70.3 % improvement. This approach presents a highly promising concept aimed at developing sustainable green waterborne polyurethane sizing agent and improving the interfacial performance of CF composite materials.

Boosting resistant starch in rice: Bacterial inulin as a metabolic and glucose uptake modulator.

Bacillus stercoris PSSR12 (B. stercoris PE), an isolate from rice field soils, was identified via 16s rRNA sequencing. The synthesis of the inulin and inulin producing enzyme (IPE) in B. stercoris PE was verified using SDS-PAGE and FTIR. This study aimed to assess the impact of B. stercoris PE treatment on in vitro inhibition of α-amylase and α-glucosidase from traditional and commercial rice varieties of South India. Additionally, the study investigated enzymatic inhibition and mRNA expression of starch synthesis genes (RAmy1a, GBSSIa, SBEIIa, and SBEIIb). Glucose transporter gene expression (GLUT1 and GLUT4) patterns were analyzed in 3T3-L1 adipocytes to evaluate glucose uptake in B. stercoris PE treated rice varieties. The application of B. stercoris PE enhanced grain quality by imparting starch ultra-structural rigidity, inhibiting starch metabolizing enzymes, and inducing molecular changes in starch synthesis genes. This approach holds promise for managing type II diabetes mellitus and potentially reducing insulin dependence.

The importance of starch chain-length distribution for in vitro digestion of ungelatinized and retrograded foxtail millet starch.

The digestibility of ungelatinized, short-term retrograded and long-term retrograded starch from foxtail millet was investigated and correlated with starch chain length distributions (CLDs). Some variations in starch CLDs of different varieties were obtained. Huangjingu and Zhonggu 9 had higher average chain lengths of debranched starch and lower average chain length ratios of amylopectin and amylose than Dajinmiao and Jigu 168. Compared to ungelatinized starch, retrogradation significantly increased the estimated glycemic index (eGI), whereas significantly decreased the resistant starch (RS). In contrast, long-term retrograded starches have lower eGI (93.33-97.37) and higher RS (8.04-14.55%) than short-term retrograded starch. PCA and correlation analysis showed that amylopectin with higher amounts of long chains and longer long chains contributed to reduced digestibility in ungelatinized starch. Both amylose and amylopectin CLDs were important for the digestibility of retrograded starch. This study helps a better understanding of the interaction of starch CLDs and digestibility during retrogradation.

Starch chain-length distributions affect the processing and digestion characteristics of proso millet starch.

Starch chain-length distributions play a key role in regulating the processing and digestion characteristics of proso millet starch. Waxy proso millet starch has higher endothermic enthalpy (13.06-16.73 J/g) owing to its higher relative crystallinity (27.83%-32.04%), while nonwaxy proso millet starch has lower peak viscosity (1.0630-1.1930 Pa∙s) and stronger viscoelasticity owing to its higher amylose content (21.72%-24.34%). Non-waxy proso millet starch exhibited two different digestion phases and its resistant starch content (18.37%-20.80%) was higher than waxy proso millet starch. Correlation analysis showed proso millet starch with longer amylopectin B1 chains and more amylopectin B2 chains exhibited excellent thermal ability and retrograde resistance, whereas proso millet starch with shorter and more amylose medium/long-chains not only reduced the digestion rate and increased the resistant starch content but also exhibited stronger viscoelasticity and excellent retrogradation properties. These results could provide more insights into efficient utilization of proso millet starch.

The Waxy Gene Has Pleiotropic Effects on Hot Water-Soluble and -Insoluble Amylose Contents in Rice (Oryza sativa) Grains.

Rice (Oryza sativa) is a cereal crop with a starchy endosperm. Starch is composed of amylose and amylopectin. Amylose content (AC) is the principal determinant of rice quality, but varieties with similar ACs can still vary substantially in their quality. In this study, we analyzed the total AC (TAC) and its constituent fractions, the hot water-soluble amylose content (SAC) and hot water-insoluble amylose content (IAC), in two sets of related chromosome segment substitution lines of rice with a common genetic background grown in two years. We searched for quantitative trait loci (QTLs) associated with SAC, IAC, and TAC and identified one common QTL (qSAC-6, qIAC-6, and qTAC-6) on chromosome 6. Map-based cloning revealed that the gene underlying the trait associated with this common QTL is Waxy (Wx). An analysis of the colors of soluble and insoluble starch-iodine complexes and their λmax values (wavelengths at the positions of their peak absorbance values) as well as gel permeation chromatography revealed that Wx is responsible for the biosynthesis of amylose, comprising a large proportion of the soluble fractions of the SAC. Wx is also involved in the biosynthesis of long chains of amylopectin, comprising the hot water-insoluble fractions of the IAC. These findings highlight the pleiotropic effects of Wx on the SAC and IAC. This pleiotropy indicates that these traits have a positive genetic correlation. Therefore, further studies of rice quality should use rice varieties with the same Wx genotype to eliminate the pleiotropic effects of this gene, allowing the independent relationship between the SAC or IAC and rice quality to be elucidated through a multiple correlation analysis. These findings are applicable to other valuable cereal crops as well.

Insight to starch retrogradation through fine structure models: A review.

The retrogradation of starch is crucial for the texture and nutritional value of starchy foods products. There is mounting evidence highlighting the significant impact of starch's fine structures on starch retrogradation. Because of the complexity of starch fine structure, it is a formidable challenge to study the structure-property relationship of starch retrogradation. Several models have been proposed over the years to facilitate understanding of starch structure. In this review, from the perspective of starch models, the intricate structure-property relationship is sorted into the correlation between different types of structural parameters and starch retrogradation performance. Amylopectin B chains with DP 24-36 and DP ≥36 exhibit a higher tendency to form ordered crystalline structures, which promotes starch retrogradation. The chains with DP 6-12 mainly inhibit starch retrogradation. Based on the building block backbone model, a longer inter-block chain length (IB-CL) enhances the realignment and reordering of starch. The mathematical parameterization model reveals a positive correlation between amylopectin medium chains, amylose short chains, and amylose long chains with starch retrogradation. The review is structured according to starch models; this contributes to a clear and comprehensive elucidation of the structure-property relationship, thereby providing valuable references for the selection and utilization of starch.

The effects of chain-length distributions on starch-related properties in waxy rices.

Normal rice starch consists of amylopectin and amylose, whose relative amounts and chain-length distributions (CLDs) are major determinants of the digestibility and rheology of cooked rice, and are related to metabolic health and consumer preference. Here, the mechanism of how molecular structural features of pure amylopectin (waxy) starches affect starch properties was explored. Following debranching, chain-length distributions of seven waxy varieties were measured using size-exclusion chromatography, and parameterized using biosynthesis-based models, which involve breaking up the chain-length distribution into contributions from five enzyme sets covering overlapping ranges of chain length; structure-property correlations involving the fifth set were found to be statistically significant. Digestibility was measured in vitro, and parameters for the slower and longer digestion phase quantified using non-linear least-squares fitting. The coefficient for the significant correlation involving amylopectin fine structure for the fifth set was -0.903, while the amounts of amylopectin short and long chains were found to dominate breakdown viscosity (correlation coefficients 0.801 and - 0.911, respectively). This provides a methodology for finding or developing healthier starch in terms of lower digestion rate, while also having acceptable palatability. As rice breeders can to some extent control CLDs, this can help the development of waxy rices with improved properties.

High amylopectin in waxy maize synergistically affects seed germination and seedling vigour over traditional maize genotypes.

Waxy maize grains rich in amylopectin have emerged as a popular food and industrial raw materials. Here, a set of waxy inbreds having recessive waxy1 (wx1) gene derived through marker-assisted selection (MAS), and their original versions were evaluated for germination, seed vigour index-I and vigour index-II, electrical conductivity (EC) and enzymatic activities viz., dehydrogenase (DH), esterase (EST), peroxidase (POX), superoxide dismutase (SOD) and α-amylase (AMY). Waxy inbreds under study possessed average 97.8% amylopectin compared to 72.4% in original inbreds. Waxy versions showed 15.2% more test weight, 4.3% increase in germination, 22.7% higher seed vigour index-I and 28.3% higher seed vigour index-II, respectively, over the original inbreds. Further, activity of DH, EST, POX, SOD and AMY of MAS-derived waxy inbreds was more than that of original inbreds, whereas EC was less in improved inbreds compared to originals. Amylopectin exhibited strong positive correlations (r = 0.69 to 0.97**) with seed germination, vigour index-I and -II, DH, SOD, POX, EST and AMY activity. However, amylopectin showed negative correlation of - 0.82** with EC. Seed germination and seed vigour indices were also positively correlated with all enzymatic activities (r = 0.58 to 0.92**). The analysis revealed that waxy inbreds possess better seed vigour and enzymatic activities over traditional inbreds. This is the first report of synergistic effects of wx1 gene on seed germination, vigour and enzymatic activities in maize endosperm.

The art and science of porous starch: understanding the preparation method and structure-function relationship.

Porous starch (PS), a modified form of starch with unique properties, is attracting substantial attention for its diverse advantages and applications. Its intricate porous structure, crystalline and amorphous characteristics, and hydrophilic-hydrophobic properties stem from pore formation via physical, chemical, enzymatic, and combined synergistic methods. Porous starch offers benefits like improved gelatinization temperature, water absorption, increased surface area, tunable crystallinity, and enhanced functional properties, making it appealing for diverse food industry applications. To optimize its properties, determining the parameters governing porous structure formation is crucial. Factors such as processing conditions, starch source, and modification methods substantially impact porosity and the overall characteristics of the material. Understanding and controlling these parameters allows customization for specific applications, from pharmaceutical drug delivery systems to enhancing texture and moisture retention in food products. To date, studies shedding light on how porosity formation can be fine-tuned for specific applications are fewer. This review critically assesses the existing reports on porous starch, focusing on how preparation methods affect porosity formation, thereby influencing the product's crystallinity/hydrophilic-hydrophobic nature and overall applicability.

Insights into wheat-starch biosynthesis from two-dimensional macromolecular structure.

Starch structure is often characterized by the chain-length distribution (CLD) of the linear molecules formed by breaking each branch-point. More information can be obtained by expanding into a second dimension: in the present case, the total undebranched-molecule size. This enables answers to questions unobtainable by considering only one variable. The questions considered here are: (i) are the events independent which control total size and CLD, and (ii) do ultra-long amylopectin (AP) chains exist (these chains cannot be distinguished from amylose chains using simple size separation). This was applied here to characterize the structures of one normal (RS01) wheat and two high-amylose (AM) mutant wheats (an SBEIIa knockout and an SBEIIa and SBEIIb knockout). Absolute ethanol was used to precipitate collected fractions, then size-exclusion chromatography for total molecular size and for the size of branches. The SBEIIa and SBEIIb mutations significantly increased AM and IC contents and chain length. The 2D plots indicated the presence of small but significant amounts of long-chain amylopectin, and the asymmetry of these plots shows that the corresponding mechanisms share some causal effects. These results could be used to develop plants producing improved starches, because different ranges of the chain-length distribution contribute independently to functional properties.