This review paper focuses on the recent advancements in the large-scale and laboratory-scale isolation, modification, and characterization of novel starches from accessible botanical sources and food wastes. When creating a new starch product, one should consider the different physicochemical changes that may occur. These changes include the course of gelatinization, the formation of starch–lipids and starch–protein complexes, and the origin of resistant starch (RS). This paper informs about the properties of individual starches, including their chemical structure, the size and crystallinity of starch granules, their thermal and pasting properties, their swelling power, and their digestibility; in particular, small starch granules showed unique properties. They can be utilized as fat substitutes in frozen desserts or mayonnaises, in custard due to their smooth texture, in non-food applications in biodegradable plastics, or as adsorbents. The low onset temperature of gelatinization (detected by DSC in acorn starch) is associated with the costs of the industrial processes in terms of energy and time. Starch plays a crucial role in the food industry as a thickening agent. Starches obtained from ulluco, winter squash, bean, pumpkin, quinoa, and sweet potato demonstrate a high peak viscosity (PV), while waxy rice and ginger starches have a low PV. The other analytical methods in the paper include laser diffraction, X-ray diffraction, FTIR, Raman, and NMR spectroscopies. Native, “clean-label” starches from new sources could replace chemically modified starches due to their properties being similar to common commercially modified ones. Human populations, especially in developed countries, suffer from obesity and civilization diseases, a reduction in which would be possible with the help of low-digestible starches. Starch with a high RS content was discovered in gelatinized lily (>50%) and unripe plantains (>25%), while cooked lily starch retained low levels of rapidly digestible starch (20%). Starch from gorgon nut processed at high temperatures has a high proportion of slowly digestible starch. Therefore, one can include these types of starches in a nutritious diet. Interesting industrial materials based on non-traditional starches include biodegradable composites, edible films, and nanomaterials. Starch makes up almost 90% of the daily calorie intake from staple foods worldwide . It affeThis carbohydrate is industrially isolated mainly from corn, tapioca (cassava), wheat, potato, sago , pea, arSeveral commercial starches originate from specific cultivars containing high-amylose starch (70% amylose) or waxy starch with a high amylopectin content. Amylose corn starch is produced by Xinfuwang New Material Technology Co., Ltd.  , and waxCanna edulis) is produced in Colombia and Venezuela  is defined in terms of the moment-ratio system as:in is the number of the measured particles and iD is their diameter. Kidney bean and canistel starches demonstrate a high value of this parameter. Ma et al. [a et al.  measureda et al.  found 65A, CC, and CB-types [Phoenix sylvestris, and ulluco tubers, or pumpkin and winter squash fruit  or quinoa starches had a high percentage of A-chains and a low percentage of B3. Similarly, the number-average degree of polymerization of amylopectin decreased with a decreasing granule size in barley starch [Chlorella, canna, and tapioca starch with high-amylose corn starch had a maximum B3 chains content  on starch is indicative of interfering with differences in starch structures, changes in the physical states of starch, and the interactions of starch polymers with other constituents and composite food matrices. This method has enabled the measurement of gelatinization temperatures, provided evidence that chemical modification has reduced or eliminated aggregation and the association of starch molecules during the cold storage of pastes, and evaluated the temperature and enthalpy of the formation of amylose–lipid complexes .Carioca beans (10.1–13.8 °C), common corn (10.4 °C), and ramon seeds (10.6 °C).The temperature at which gelatinization occurs and the heat enthalpy are closely connected to the starch structure and molecular arrangement. Furthermore, the amylose content could also influence the enthalpy of gelatinization. The thermal data found in recent papers are in Euryale ferox seeds and sago pith waste fiber demonstrated the highest enthalpy.The gelatinization endotherm is associated with the gelatinization of the crystalline amylopectin lamella, but not necessarily to the proportion of the crystalline area, which corresponds to the amylose: amylopectin ratio and the distribution of amylopectin chains in the granule . A higheretH) reflects the energy required to melt the recrystallized starch structure and starch molecular double helices formed during long-term storage. There are not enough data for a comparison of ΔretH. The highest value was found for sand rice starch , while Phoenix sylvestris starch demonstrated the lowest value.The retrogradation enthalpy (Δw/w) to 16.67% (w/w)—see As for the pasting properties, viscosity describes the magnitude of the friction generated by the mutual movement of starch granules after gelatinization . A compaThe peak viscosity (PV) of starch indicates the maximum swelling caused by heat in the presence of water . When weBreakdown (BD) equals PV minus trough (minimum) viscosity . BD reprHigh final viscosities (FV) were found in Turkish bean and winter squash starch. On the contrary, this was low in waxy amaranth. Setback (SB) equals FV minus trough (minimum) viscosity . SB indigm— the weight of the gel (g), stm—the weight of the starch sample (g), and solm—the soluble weight (g).The extent of granule swelling is determined by measuring the swelling power , which is reported as the ratio:SP measures the hydration capacity, because food-eating quality is often associated with the water retention in the swollen starch granules .Solubility  is the percentage of molecules leached from the starch granules after swelling under specific conditions:The SP and SW values provide information about the degree of interaction between the chains present in the crystalline and amorphous regions of the starch granules and are correlated with amylose content . A higheSP. The results of the swelling power solubility and water absorption of different starches, at temperatures ranging from 60 to 80 °C, are presented in Agriophyllum squarrosum, and Phoenix sylvestris at 80 °C. All starches showed an increasing swelling power at increasing heating temperatures because of gelatinization.The water absorption index  is defined as:Phoenix sylvestris showed the highest SW values measured at 80 °C, while anchote, ginger, and foxnut starches demonstrated the lowest SP values.Starches of canna, quinoa, black bean, and Frozen foods containing starch can be repeatedly freeze–thawed during storage and transportation. Thermal fluctuations in foods during transport cause water melting and recrystallization, lowering their quality and acceptability. Simultaneously, the starch recrystallizes during cooling, resulting in water separation known as syneresis. Syneresis is an undesirable phenomenon that results in a poor starch gel texture .Chemical modifications such as acetylation and hydroxypropylation can positively influence freeze–thaw stability. These processes have also been tested on non-traditional starches .Zarroug et al.  studied -the lower the temperature of the refrigeration, the higher the syneresis,-the longer the storage time, the higher the syneresis,-the syneresis of acorn starch is comparable to corn starch,-the syneresis of sweet potato starch and Turkish bean starch is lower than that of corn starch.Hussain et al.  publishePhotinam et al.  reportedAccording to Liu et al. , native As the principal energy source of the human body, the digestibility of starch is an important parameter in the quality assessment of food. During starch digestion, glucose is released. Thus, starch digestion is closely associated with the postprandial blood glucose and insulin levels in humans . It is iThe amorphous area of the starch molecule is more amenable to acid attacks, being more easily and quickly hydrolyzed . HoweverThe lower digestibility of starch results in lower obesity and microbial formation of short-chain fatty acids in the human gut, thus having healthy effects . EnglystThe lower in vitro digestibility of legume starches than that of cereal starch might be due to their high amylose content (because of their lower susceptibility to enzymatic hydrolysis), the absence of surface pores on the granules, large amounts of B type crystallites, strong interactions between amylose chains, intact cell structures enclosing starch granules, and large amounts of soluble dietary fiber .The starch molecules in freshly gelatinized starches are amorphous and susceptible to digestive enzymes . Transglcooked lily starch (20%) and a high RS content was found in lily starch (>50%) and unripe plantains (>25%). The highest SDS was found in the native starch of annatto and Euryale ferox (>35%). A very high SDS content was found in the thermally (80–100 °C) processed starch of gorgon nut.However, most of these starches are not eaten in their native form. A low RDS is maintained in tC and ∞C (mg/mL) are the digestibility of starch when the hydrolysis time is t (min) from 0–k indicates the kinetic coefficient of the starch digestibility (min−1). Liu et al. [∞C at 42.6% and k at 0.029 min−1 for chestnut starch, ∞C at 64.0% and k at 0,0264 min−1 for Kutki millet starch, and ∞C at 66.5% and k at 0.0297 min−1 for adzuki bean starch, resp., compared to ∞C at 65.6% and k at 0.0332 min−1 for corn starch. Mahajan et al. [Besides the evaluation of RDS, SDS, and RS, simulations of starch digestion in the small intestine were performed using various evaluation models ,87. The u et al. , and Wu u et al.  evaluaten et al.  also lisHSm (g)—the weight of hydrolyzed starch and ISm (g)—the weight of initial starch [The simple magnitude is the degree of starch digestion or enzyme hydrolysis (%) defined as:l starch ,115. TheFourier-transform infrared spectroscopy (FTIR) is a powerful tool for characterizing the structure of starch and starch-based composite films ,89,116. −1 represents the stretching vibrations of O–H bonds in hydroxyl groups and overlap narrow weaker bands at 2928–2931 cm−1 attributed to the stretching vibration of the C–H bond in glucopyranosyl residues. The bands observed at 1153, 1086, 1012, and 930 cm−1 in the spectral region, often assigned as “sugar region”, are attributed to C–O–C, C–O, and C–C stretching and C–O–H bending vibrations in the glycosidic linkages and pyranoid rings [−1 and 767 cm−1 are characteristic for 1,4-α-glycosidic bonds in starches [−1 arises from C1α–H and CH2 deformations and is indicative of the α-anomer of glucopyranose [−1 and below arise from the skeletal modes of the pyranoid ring of α-glucopyranose [The observed FTIR spectra are similar for all studied starches ,117. A bid rings ,25. Sevestarches , and thepyranose . Finallypyranose .−1 (twisting vibration of bound water), 1640–1650 cm−1 (HOH scissoring vibration), and 3500 cm−1 (HOH stretching vibration). In hydrated starches, the band near 2100 cm−1 can be attributed to the free water content [−1 [Water molecules play a crucial role in starch structure  and cont content ,121 and tent [−1 .−1 is related to the amorphous phase [−1 and 1047 cm−1 correspond to the crystalline region of the starch [−1/1022 cm−1 and 995 cm−1/1022 cm−1 quantitatively characterizes the degree of short-range order and the proportion of double helices in starches, respectively [Agriophyllum squarrosum) and barley starches, the ratio of band intensities at 1047 cm−1/1022 cm−1 confirmed that polysaccharide chains are rearranged when heated under a low humidity, equal to or below 20%, but at a higher humidity, reaching 30%, short-range ordered structures began to be disrupted [The bands in “sugar region” are sensitive to hydration and the crystalline/amorphous stay of starches. The IR band at 1022 cmus phase , while te starch ,89. The ectively . For sanisrupted . The menisrupted ,115,123.−1 and at 2940 cm−1 increased after the partial acidic hydrolysis of achira (Canna edulis) starch [2 vibrations and were sensitive to hydrogen bonds involving starch hydroxylic groups. The increase in these bands may indicate an enhanced number of free CH2OH groups due to the hydrolysis of 1,6-α-glycosidic bonds (unbranching).The intensities of the bands at 1200–1400 cm) starch . These b−1 and some disturbances at 1060 cm−1, probably due to stretching of the C1–O5 bond in the pyranoid ring [−1 assigned to the C=O stretching vibration of the carbonyl group [−1 due to vibrations of new carbonyl and carboxylate groups [−1/1022 cm−1 [−1 [2 stretching vibrations at 2860 and 2926 cm−1 also appeared and increased with the length of acyls. A new IR band of phosphorylated starches at 1417 cm−1 corresponded to the P=O stretching vibration [The FTIR spectra of chemically oxidized cassava starch showed a carboxylate antisymmetric stretching band at 1600 cmoid ring . These syl group . The oxie groups . The dif022 cm−1 . The FTIcm−1 [−1 . Two IR ibration .−1 and 2917 cm−1 assigned to the antisymmetric and symmetric stretching of the CH2 groups in the long-chain fatty acids. In addition, a weak band at 1383 cm−1 assigned to the CH3 bending vibration was also found [−1 (amide I), 1545 cm−1 (amide II), and 1240–1350 cm−1 (amide III) are attributive to the vibrations of amide groups in proteins, which are often present in starch preparations [The FTIR spectra of adzuki bean starch–fatty acid complexes demonstrated two narrow bands at 2850 cmso found . Additioarations ,25.−1, corresponding to O–H stretching in glycerol, starch, and water [−1 assigned to S=O stretching, 3,6-anhydrogalactose, and galactose-4-sulfate vibrations, respectively [−1, indicating the asymmetric stretching vibration of the CH3 groups of the acetyl and methyl esters in cactus (Opuntia ficus-indica) and acacia  gums in a mixture with various starches. The IR band observed at 1721–1728 cm−1 indicates the C=O stretching vibration of the carboxylic acids and esters in polysaccharidic gums. The bands at 1415–1400 and 1310–1230 cm−1 corresponded to the symmetric stretching of the COO- of the carboxylate groups and C–O stretching of the carboxylic acids in the gums, respectively.Composite starch-based edible films often contain proteins, lipids, aromatics, and other polysaccharides. FTIR spectroscopy is able to detect the presence of these compounds via characteristic vibration bands. Starch-based film represents specific bands at 3326 cmnd water . The FTIectively  observedIn addition to FTIR, Raman spectroscopy is also widely used in the structural analyses of starches, for example in the characterization of the structural changes in retrograded ,127, mic−1 correspond to C–H stretching, C–H, CH2, and C–O–H bending, C–O–H, C–C–H, and O–C–H bending, C–O and C–C stretching, C–C–H and C–O–C bending, and pyranoid ring skeletal vibrations, respectively. In amorphous starch, Raman bands, except those at 2910 and 478 cm−1, were weak and almost disappeared at low frequencies. The Raman intensity varies directly with the size of the crystalline region in starch. The Raman bands for B-type starch  were less intense than those for A-type starch , and the most of these bands shifted to lower wavenumbers. De Gussem et al. [−1 compared to the corresponding bands of amylopectin at 410, 477, 769, and 865 cm−1. The skeletal vibration band of starch near 477 cm−1 is one of the most intense in the Raman spectrum and can be used as a marker to detect starch in mixtures with other polysaccharides [The Raman spectra of starches are sensitive to the presence and type of crystalline structure, degree of substitution, and relative amounts of amylose and amylopectin. The band intensity in the Raman spectra for starches depends on their structural features . The typm et al.  observedcharides .2O stretching bands [−1 assigned to the vibrations of trimethylammonium substituents. The ratio of the band intensities at 761 cm−1/938 cm−1 (reference starch band) can quantify the degree of substitution. Raman spectroscopy was applied to determine the degree of succinylation in modified starches [Since Raman spectroscopy is less sensitive to the contribution of water and polar groups, it is more prospective in the analysis of aliphatic moieties in substituted starches. For example, no significant changes were observed for cationic quaternary ammonium starches in the FTIR spectra due to the overlap of the strong OH and Hng bands . In contstarches .1H NMR has been applied to the determination of the degree of substitution of acetylated and other acylated starches [1H and 13C NMR [Nuclear magnetic resonance (NMR) spectroscopy is a powerful tool in the structural analysis of natural and modified polysaccharides, including starches. Applications of this method in the characterization of starch systems were reviewed by Zhu . Variousstarches ,137,138, 13C NMR .13C CP/MAS NMR spectra can evaluate the molecular organization of starch samples [2OH groups [13C CP/MAS NMR spectra deconvoluted by subtracting the scaled NMR spectrum of the amorphous starch to zero intensity at 84 ppm yield the ordered and amorphous phases of the starch samples [13C CP/MAS NMR spectroscopy. The obtained results confirmed that the structure of the modified starch changed from A-type double helical to V-type single helical and the short-range crystalline structure decreased as well.Solid-state  samples ,63,89. B samples . After tH groups . For nat samples ,141,142.1H NMR can evaluate the molecular mobility and water distribution in the starch–water systems [Time domain  systems . This meNative starch may be modified in numerous ways, for example, physically, chemically, or via an enzyme action.Pregelatinized starch originates when starch slurry is heated by steam . It is aPinhão starch. There was an increase in the gelatinization temperature and a decrease in the ΔH after treatment. HMT changed the short-range ordered structures, shifted the X-ray pattern from C-type to A-type, and reduced the relative crystallinity. Akinyosoye and Nwokocha [Treculia africana starch. As the moisture level of HMT increased, the swelling decreased, and there was an increase in amylose leaching. Pasting temperature and retrogradation increased, and breakdown decreased with HMT. Thus, HMT strengthened the granule structure, resulting in starch which can withstand severe processing conditions without viscosity loss. Similarly, Wu et al. [Biduski et al.  found thNwokocha  investigu et al.  tested tu et al. , swollenThe ANN treatment applied to ulluco starch proved to be an efficient method for improving the physicochemical parameters such as solubility, water retention capacity, and the texture of the gel formed, which are valuable properties .Natural starch can be modified in various ways to produce acid- or enzyme-modified, oxidized, cross-linked, esterified, or etherified starch, converted into cationic derivatives, or subjected to pyroconversion . These mPartial hydrolysis is a way for the fabrication of acid- or enzyme-modified starches. This process is suitable for the preparation of porous starch, e.g., from banana  or lotusTung et al.  used a hPhosphorylation and acetylation change the functional and physicochemical properties of starches. The hydroxypropylation of non-traditional starches enhances swelling capacity, viscosity, clarity, and freeze–thaw stability ,106.Also, a combination of physical and chemical modification was tested, e.g., ozone oxidation together with high-pressure homogenization  or high Generally, starch can be used as an additive, gelling agent, thickener, fat replacer, and emulsifier in crunchy foods, baby foods, and controlled release systems, and as a flavor enhancer and coating agent in many industrial foods, feeds, and drugs to improve their functional and sensory characteristics ,112,145.The development of new starch raw materials can be used for developing healthy foods required by consumers, thereby enhancing the value-added application . They arResistant starch (not accessible to human digestive enzymes) has beneficial effects for health; it positively influences the digestive tract and its fermentation in the colon causes the formation of short-chain fatty acids and the lowering of the pH. It further increases fecal bulk, protects against colonic cancer, improves blood cholesterol levels, assists glucose tolerance control in diabetes, and causes lower blood lipid levels. However, the current RS/dietary fiber intake by Europeans is insufficient. Therefore, it is suitable to add it to food. According to Du et al.  and YuanChenopodium, and jackfruit [Some papers have solved the utilization of starch from wastes, e.g., coming from seeds of annatto , avocadoackfruit ,54,83,98ackfruit .In recent years, the importance of high-amylose starch has grown due to its expanded range of functionalities compared to native starch. These functionalities include nutrition, food processing, medicine, and industrial use .An outstanding group of starches covers starches having small granules. Cow cockle starch (0.3–1.5 μm), rice starch (2–8 μm), and amaranth starch are suitable as fat substitutes in frozen dessert and mayonnaise; similarly, acorn starch has been used in custard because the small particle starches could form a smooth creamy fat-mimetic texture ,145,147.Nanomaterials exhibit properties that are distinctive and qualitatively different from large-size particles. Starch nanoparticles (SNPs) developed from mango starch, mungbean, and water chestnut starch are comparable with wheat or potato nanoparticles. SNPs also have great potential for papermaking use, surface sizing, coating, and in paperboard as a biodegradable adhesive instead of petroleum-based adhesives. All the tested SNPs are non-cytotoxic and can be safely used in biomedical applications . NanocomEscherichia coli, Salmonella typhimurium, and Staphylococcus aureus, which results in a high potential for use in active packaging in food industries. Nanocomposite films processed with sugar palm starch, sugar palm nanocrystalline cellulose (SPNCC) with cinnamon essential oil have similar effect [Plastic-based synthetic polymers widely used in modern food packaging contribute to plastic-waste end-products, which accumulate in and pollute terrestrial and aquatic environments. Thus, there are ways of reducing the use of and dependence on them . Low cosr effect .Edible biodegradable films (EF), intended for primary food packaging use, are made from food-grade and biodegradable polymers. Therefore, edible films are used to wrap the food product to inhibit the moisture uptake, extend the shelf life, and are consumed together with the food. Consequently, EF can generate almost zero waste after disposal . EFs fabSeveral non-conventional starches have technological advantages over common starches for specific applications in the cosmetic and paper industries . In addiNowadays, the research focuses on novel starch sources in the context of food security, sustainable production, the use of by-products, e.g., coming from annatto and avocado, their regional availability, and technological advantages over traditional starches.Published numerous data like shape, size, and crystallinity characterize non-traditional starches. Small-granule starches demonstrate unique properties due to a higher water affinity, larger surface-area-to-volume ratio, and low sedimentation coefficient. Because of their smooth texture, they can be utilized as fat substitutes in frozen desserts, mayonnaise, and custard. These starches have shown an excellent compatibility in biodegradable plastics. They also can be applied in their native or modified form as adsorbents.Phoenix sylvestris with a high swelling power.The practical use of starches in the food industry requires knowledge of the thermal and pasting properties of the suggested starches. The low onset temperature (To) in acorn starch is associated with the costs of the industrial processes in terms of energy and time. The requested high retrogradation enthalpy was characteristic of sand rice starch, the smallest one for starch from Starch plays a crucial role in the food industry as a thickening agent. Starches from ulluco, winter squash, bean, pumpkin, quinoa, and sweet potato demonstrate a high peak viscosity (PV); on the other hand, waxy rice and ginger starch had a low PV. Low setback (SB), indicating recrystallization during the cooling of starch paste, was found in waxy varieties and chestnuts. Turkish bean, sand rice, and quinoa showed the highest SB.New food trendy categories comprise plant-based food e.g., snacks and food containing “clean-label” or resistant starch. Native, “clean-label” starches from new sources could replace chemically modified starches, due to their similar properties compared to common commercial modified ones. Furthermore, their production does not produce chemical by-products. These techniques for preparing “clean-label” starches include pregelatinization, microwave treatment, annealing, heat moisture treatment, high-pressure homogenization, ultrasound, or ultrasound combined with plasma treatment.As fiber, resistant starch (RS) has beneficial effects on health. It positively influences the digestive tract and its fermentation in the colon protects against colonic cancer. RS improves blood cholesterol levels, assists glucose blood level control in diabetes, and causes lower blood lipid levels. A high RS content was found in gelatinized lily starch (>50%) and unripe plantains (>25%), and starch from indigenous Himalayan folk rice has a amylose content higher than 32%. The low content of rapidly digestible starch is maintained in cooked lily starch (20%). Thermally processed starch from gorgon nut showed a very high SDS. Therefore, these starches are suitable for a healthy diet.However, chemical modifications of non-traditional starches are still being tested, including oxidation, phosphorylation, acetylation, hydroxypropylation, and a combination of physical and chemical approaches. Spectroscopy methods are powerful tools for the characterization of the purity, structure, and physical properties of starches, their derivatives, and composite materials based on them. In addition to X-ray diffraction, FTIR, Raman, and NMR can evaluate the ratio between the crystalline and amorphous forms of starch.Native and modified non-traditional starches can be used also outside the food industry. Materials from sugar palm, cassava, corn, and others have been discovered to be suitable as polymer composite reinforcements. Edible foils fabricated from starch polysaccharides are one of the primary genuine forms of biopolymer materials for food packaging due to their strength, processing, product adaptability, rheological properties, and performance; they are environmentally friendly and cost-effective. A relatively new progressive modification of starches enables the creation of nanomaterials, including nanoparticles/nanofillers, nanofibers, or nanocomposites, providing perspectives for various fields of industry.


Starches with an amylose content of over 30% conditionally belonged to the “ae” genotype. The starches of the su genotype had fewer starch granules than other investigated genotypes. An increase in amylose content in the investigated starches, accompanied by a decrease in their thermodynamic melting parameters, induced the accumulation of defective structures in the starches under study. The thermodynamic parameters evaluated for dissociation of the amylose–lipid complex were temperature (amlT) and enthalpy (amlH); for the su genotype, temperature and enthalpy values of dissociation of the amylose–lipid complex were higher than in the starches from the “ae” and WT genotypes. This study has shown that the amylose content in starch and the individual features of the maize genotype determine the thermodynamic melting parameters of the starches under study.To understand the relationship between the genotype of maize plants and differences in their origin and the ploidy of the genome, which carry gene alleles programming the biosynthesis of various starch modifications, the thermodynamic and morphological features of starches from the grains of these plants have been studied. This study investigated the peculiarities of starch extracted from subspecies of maize (the dry matter mass (DM) fraction, starch content in grain DM, ash content in grain DM, and amylose content in starch) belonging to different genotypes within the framework of the program for the investigation of polymorphism of the world collection of plant genetic resources VIR. Among the starch genotypes of maize studied, four groups comprised the waxy ( Cereal crops are the primary sources of starch. In addition, tuber crops , legumes, and some unripe fruits, such as bananas and mangoes, contain starches . As knowGBSS) is crucial for amylose synthesis [wx) are characterized by the absence of GBSS accompanied by the accumulation of only amylopectin in the complete absence of amylose. The amylose type of grain endosperm (ae) is characterized by a violation of SBEIIb accompanied by the effect of the accumulation of weakly branched polysaccharides and the accumulation of amylose in starch [su) genotype with sucrose synthases (SS) generating phytoglycogen as an amylopectin precursor has a deficit of such enzymes as isoamylases and pullulanases disturbing branching in starches. The high sugar and low starch contents in sweet sorghum and maize indicate the high activity of this enzyme. Grains of these plants, as a rule, are wrinkled with a low DM concentration [n) to tetraploid (4n), the number of alleles of the su2 genes that control the synthesis and accumulation of sugars in the endosperm of grain with the genotype su2su2su2 (2n) to su2su2su2su2su2su2 (4n) is increasing. Accordingly, the number of dominant alleles of the Su2 gene (wild type) in ordinary corn will affect the synthesis of normal starch in the same proportions as Su2Su2Su2 (2n) Su2Su2Su2Su2Su2Su2 (4n) without the accumulation of sugar in the maize grain. The findings of investigations into the grain chemical components affecting the taste qualities of tetraploid and diploid sweet maize have shown that an increase in the number of genes is accompanied by a change in the quantitative, rather than qualitative, biochemical profile characteristics [Structural and thermodynamic properties of starch were shown to be affected by the conditions of starch-yielding plant cultivation and by the presence of specific genes involved in the biosynthesis of starch polysaccharides . The actynthesis , while mynthesis . Three kynthesis . The genn starch ,24. The n starch ,26. Mutan starch ,27. The ntration ,29,30. Ieristics . StudiesOur study of polymorphism of plant genetic resources of the world VIR maize collection determined standard values of thermodynamic and structural features of starches isolated from maize genotypes of different origins and genomic ploidy, carrying alleles of genes programming biosynthesis of various starch modifications. Tetraploid genotypes found in the standard study of the diversity of the VIR world maize collection resources allowed a comparative analysis of some starch properties with diploid ones. The prospect of possible regulation of starch quality by changing the number and ratio of different genes responsible for starch synthesis is a promising direction in genetics and breeding. It is also possible to adjust the granule morphology and crystallinity of extracted starches at the genetic level. As a rule, not only one or two genes but also a gene network regulates starch morphological features, including its physicochemical properties. Thus, the genotypes containing genes determining the starch structure and physicochemical properties will differ in all or some parameters, representing optimal raw materials for different industries. In this regard, the purpose of this study was to evaluate the characteristics of the products of processing grain for starch, as well as to understand the relationship between the thermodynamic and structural features of maize starches from samples of the VIR world collection with different ploidy and genotype of maize plants from which starches are extracted.su2 gene. Tetraploid maize (2n = 40)  are closely related genotypes that differ from each other only by a mutation in the su2 gene, from diploid maize (2n = 20) by a double set of chromosomes, and a great size grains and germ (wx), sugar (su), WT (wild), and conditionally high amylose (“ae”). Maize samples with an increased amylose content of more than 30%  in our study are conditionally assigned to subspecies of amylose type “ae” since the increased content of amylose starch does not allow us to classify these samples as normal, but they are not yet typically high amylose.The maize material was reproduced for the research in 2017 in a foothill area at the Kabardino-Balkarian Research Institute of Agriculture. The breeding site was within the piedmont zone of the North Caucasus, at the watershed dividing the rivers Urvan and Nalchik. Meadow chernozems mainly represented soils. Cultivars and hybrids were reproduced under paper cages using the technique of cross-pollination within a population on a plot. Harvesting was manual, scheduled as soon as the ears reached full ripeness. The maize seed material was represented by 11 accessions of non-transgenic flint, dent, and sweet maize subspecies from the global collection held by the N.I. Vavilov All-Russian Institute of Plant Genetic Resources , selected for their high starch content in grain as part of a program for the study of polymorphism of plant genetic resources of the VIR world collection. Among the selected accessions were cultivars  of Russian and foreign origin and a hybrid (Otbornyy 150SV) developed domestically. The accession Baksanskaya sakharnaya was isolated from Populyatsiya MRPP22 as a mutant for the and germ . In our Aspergillus niger  was employed to facilitate the crashing of the specimens.In the morning hours, specimens of maize seedling rootlets prepared for chromosome analysis were fixed in acetalcohol (Carney’s fixative solution) in the phase of three-day-old plantlets. Chromosome staining was performed according to the Feulgen protocol, using the Schiff reagent after hot hydrolysis in 1N HCL at 60 °C for 10 min; beyond that point, chromosomes were stained at room temperature for 1 h with the subsequent triple washing of specimens in sulfur water . Macerat2 was ~0.20%) and infused for 48 h at 48–50 °C. The maize extract was separated from the grain ). The ground grain was thoroughly washed from the crusher with 100 mL of distilled water. The maize germ was isolated from the grain matter, rinsed to remove free starch through a 0.5 mm metal sieve mesh with 100 mL of water, and dried in a drying cabinet at 50–52 °C. After that, the grain matter separated from the germ underwent fine grinding in a Waring LB20ES laboratory blender  at 40 °C for 5 min with a rotational speed of 3000 rpm. The resulting fine substance underwent sifting through six 70 μm nylon sieve meshes. The grain matter left on the sieve surface underwent multiple washing with distilled water  until the rinsing water had become starch-free, verified by the iodine test. The resulting cellulose contained different mass fractions of the ‘bound’ starch and then dried at 55–58 °C until the constant weight. The starch/protein suspension underwent centrifugation  for 30 min at 6000 rpm, and the starch and protein (corn gluten) sediment underwent further separating by washing; repeated centrifugation separated the gluten layer from the starch. The gluten suspension separated from starch was centrifuged  for 20 min at 6000 rpm. The obtained starch and gluten were dried separately at 50–55 °C for 24 h with subsequent measuring of the product yield and the mass fraction of starch in by-products, including the same in the rinsing water. The resulting starch was studied using the described physicochemical methods.Starch was isolated from maize grain using the modified technique offered by Adkins and Greenwood . Maize gGrain characteristics  were assessed in compliance with the standard European techniques .Amylose content was measured colorimetrically according to the described technique . The resChromosome numbers of the specimens were calculated using transmitted light microscopy. For each maize accession, the number of chromosomes in a somatic cell from the rootlet was counted in 15 metaphase plates of the crushed specimen under an Olympus CX43 microscope  using 1600× zoom with immersion.Light microscopy was used to analyze the morphology of the starch granules under study. Starch was placed on a microscope slide and stained with one drop of Lugol’s iodine (a solution of potassium iodide with iodine in water) . The preMorphological changes of the starch granule were investigated with scanning electron microscopy (SEM), obtaining photos of starch granules using a scanning electron microscope Mira3 LMU  at room temperature in the conditions of a high vacuum with an accelerating voltage of 500 V.The size and morphology of the maize starch granules were measured or estimated using an extended suite of morphometric algorithms under the “Altami Studio” licensed software GUI. Besides the standard granulometric characteristics, the ellipticity factor was analyzed and compared. The ellipticity factor was calculated using the abovementioned software as the ratio between the starch granule squares and squares of the ellipses with equivalent moments of inertia, taking the ellipticity coefficient to be unity in the case of elliptic approximation.C was calculated as in [θ range from 6° to 36°. In this equation, I(2θ) is experimental XRD intensity of a starch sample, bI(2θ) is a baseline intensity which is assumed to be represented by a straight line passing through experimental intensity points at 2θ ≈ 6°, and aI(2θ) is the hypothetical intensity from the starch amorphous part. aI(2θ) was determined on the basis of experimental XRD pattern of amorphous starch obtained by milling of a normal maize starch (amylose content 20–22%) in a ball mill for 8 h [θ ≈ 21°.X-ray diffraction (XRD) study of maize starches was carried out in an air-dry state using the modernized X-ray diffractometer HZG4  operated in Bregg-Brentano geometry with diffracted beam graphite monochromator and scintillation counter (CuKα radiation). X-ray diffraction patterns for each starch sample were recorded three times and then averaged. The total starch crystallinity ed as in  with the for 8 h . For calw/w) water dispersions of the investigated maize starches were measured using the method of highly sensitive differential scanning calorimetry (DSC) on a DASM-4 microcalorimeter . The volume of the sample under study was 0.5 cm3 in a closed capsule. Measurement conditions were in the temperature range of 20–120 °C, at the heating rate of 2 °C/min, and under the constant pressure of 2.5 atm, repeating DSC measurements for each sample three times. The excess heat capacity scale was calibrated using the Joule–Lenz effect for each experiment. As shown earlier, under the given conditions, there was no need to consider thermal lags or the sample treatment duration in the calorimeter capsule [Thermodynamic melting parameters for 0.3% ( capsule . Deionizm) was calculated for the anhydroglucose residue (162 g/mol). As a rough approximation, the starch melting process can be considered a quasi-equilibrium one [Mean values of thermodynamic melting parameters for the crystalline lamellae were identified in starch out of no less than three concurrent measurements. The melting temperature value matched the maximum heat capacity peak on the thermogram. The value of experimental enthalpy corresponded to the area under the peak of the excess heat capacity curve as a function of temperature. Molar enthalpy of melting  were calculated as described earlier [m is the melting temperature for a crystalline lamella in starch; Cp is the maximum on the ordinate of the heat capacity peak on the thermogram; ΔCp exp is the difference in heat capacity values between native and molten state. The cooperative melting unit (ν) and thickness of the crystalline lamella (Lcrl) were calculated as described in the published source [m is the experimental enthalpy of melting for the crystalline lamella and ΔHvH is the Van’t Hoff enthalpy of melting.The Van’t Hoff enthalpy values , the free surface energy (γi), and enthalpy (si) for the faces of crystalline lamellae, are possible to obtain from the Gibbs–Thomson equation for semicrystalline polymers [0m and ΔH0m are the melting temperature and enthalpy, respectively, of a hypothetic crystal of unlimited size  or such crystal whose role of free surface energy may be ignored in favor of bulk energy; γi is the free surface energy for the faces of crystalline lamellae; pcrl and Lcrl are the density and thickness of a crystalline lamella, respectively.The thermodynamic melting parameters of crystalline lamellae, namely the surface entropy , conditionally to “ae” (White Flint and Gornaya chechenskaya) and su types  (ae” is based on the fact that the content of amylose starch was higher (above 30%) than that of ordinary corn (up to 25%), but did not reach the value of high-amylose maize (above 50%). The remaining maize samples  are classified as wild (WT) types based on the starch content in the grain corresponding to the content in ordinary maize. The cytological analysis resulted in the finding that accessions Baksanskaya sakharnaya and Populyatsiya MRPP22 have the tetraploid chromosome number (4n), whereas all other accessions have the diploid (2n) number of chromosomes in their genome (sugary2 gene in the homozygous recessive su2su2su2su2 (Baksanskaya sakharnaya) and dominant Su2Su2Su2Su2 (Populyatsiya MRPP22) states. Grains carrying alleles of the su2, wx, ae gene in the genome showed slight phenotypic differences mainly pronounced in sweet maize  . The conr genome . Tetraplet maize I.wx) and those with the amylose one (“ae”) demonstrated similar values of the DM mass fraction; with this, the amylopectin genotype possessed higher amounts of minerals compared with the amylose genotype reflected in the mass fraction level of ash content in grain. Both genotypes had close to similar levels of starch content in grain: from 69.3 to 73.7%. Here, the starches from the amylose genotype contained 32.0–38.8% of amylose, and those from the amylopectin genotype 0–15% of amylose. The sugary genotype (su), represented by tetraploid accession, showed the highest value of the DM mass fraction (from 92.8% up to 93.3%), the lowest starch mass fraction (58.8–65.9%) in grain DM, and the ash mass fraction of 1.5–1.7% in grain DM. Concurrently, the minimum mass fraction of starch from grains of su genotypes was accompanied by the amylose fraction in starch equal to 25.5–27.7% of grain DM. These data confirmed the assumption that the number of the main polysaccharides in starch (amylose and amylopectin) and their ratios are determined by the plant genotype. Nevertheless, there were some differences in the studied parameters within each of the four groups with the sugary, amylose, amylopectin, and wild endosperm types.wx, “ae,” and WT genotypes showed high starch content after profound grain processing (from 69.3 to 75.3% of dry matter). The content of the su genotype starches in the grains with corresponding genotypes was high enough for sweet maize, reaching 59.3–65.9% of the grain dry matter, depending on the variety.Maize accessions with the ae” maize genotype. Starch extraction from the tested maize cultivars White Flint and Gornaya chechenskaya showed an 87–90% ratio, i.e., more than other studied accessions with different genotypes. However, grain samples of the wx type had a starch extraction ratio of 83.1–84.6%, which is high enough for the processed grain of waxy maize. Note that the starch yield from grains of the su genotype was the smallest among the investigated maize cultivars. Little starch granules, non-typical for maize grains, explained a low ratio of starch extraction from the su-type grains , so in this case, the losses in by-products tended to increase significantly.The results of starch extraction from the grain with different maize genotypes during laboratory-based processing  demonstrwx genotype did not change significantly and amounted to 1.4–1.7% of dry matter. The mass fraction of amylopectin in starch from maize with the wx genotype was close to 100%.The mass fraction of ash in the grain samples under study with the ae” genotype . Among the cultivars with the su genotype, the mass fraction of amylose in starch in variety Ranyaya Lacomka was 27.7%, the highest in comparison with other investigated varieties with the su genotype.The highest mass fraction of amylose in starch among the investigated maize starches was for starch of the “su genotype was observed, and the highest starch content was in the grains of the wild genotype with the following set, depending on the plant genotype for the increase of amylose content in starch: wx ≤ WT ≤ su ≤ “ae”.Thus, the lowest starch content in the grains of the wx-type starch, but the “ae” images and su genotypes contained both blue macromolecules of amylose and brown ones of amylopectin. These micrograph images also demonstrated that the starches isolated from different maize genotypes had different granule sizes. It is shown that the granules of the large and small starch fractions contained different quantities of amylose and amylopectin molecules. According to the published data, the intensity of crystalline reflexes is higher in starch samples with larger granules than in those with smaller granules. Therefore, these factors may affect the structural arrangement of starch granules and the thermodynamic parameters of starch melting [ melting .The starch morphology in the granules was analyzed by scanning electron microscopy (SEM). The size and shape of maize starch granules were evaluated both using direct morphometric parameters and relative values ). wx genotype characterized the maize starch granules of the “ae” genotype, while the proportion of irregular or cubic granules in the starch of the “ae” genotype was higher than in the starch of the wx genotype. The size of the starch granules WT genotype had higher values of geometric parameters compared to the wx genotype of starch granules and lower values of the same parameters than the “ae” genotype. Simultaneously, the proportion of irregular granules in the starches WT genotype was lower than in the “ae” genotype of starch but slightly more than in starches of the wx genotype. It should be emphasized that starch granules of the su genotype had significantly smaller sizes compared to granules of the “ae” and wx genotypes, which may be due to the individual genotypic features of this genotype. Hence, the following settings could be drawn up according to the increase of geometric parameters for investigated starches depending on the plant genotype: su ≤ wx ≤ WT ≤ “ae”. Note that the starch of wx type had a lower proportion of irregular granules, and it is possible to imagine the increase in the number of irregular starch granules in the investigated starches dependently on the plant genotype as follows: “ae” ≥ su ≥ WT ≥ wx. Note that our results on the granule size of the su maize genotype correlated with the literature data [su type from those isolated from the wx and “ae” genotypes.Note the different sizes of maize starch granules from plants with different genotypes . Thus, fure data . It is pwx-genotype) and normal maize  starches [ae-genotype) [su genotypes than for starches of wx-genotype  are depicted in wt, su, and ae, correspondently) (C = 26–30%). The higher crystallinity of wx-genotype starch was associated with the highest amount of long- and short-chain amylopectin in this starch compared to starches of other genotypes [wx-starch, this starch would melt at a higher melting temperature. The crystallinity of su-genotype starch was less than for other investigated starches (wt and wx genotypes.starches  and B-poenotype) ,46. It cenotype) ,46,47. Tstarches ,46,48. Tenotypes . Considestarches . Probablwx-genotype) maize starch extracted from varieties Mais agestano and Mestnaya because this starch was amylose-free. wx to “ae” genotype decreased with an increase in amylose content, while in the “ae”-genotype starches with the amylose content of 32.0 and 38.0%, the amylopectin melting enthalpy values virtually matched the respective values in amylopectin starches from the wx genotype. The higher transition temperature is associated with higher crystallinity with provided structural stability [wx genotype (varieties Mais agestano and Mestnaya) showed the highest value of the melting temperature among the investigated maize starches (c) and onset temperature (T0) of amylopectin crystalline lamellar melting for investigated maize starches may be due to the changes in the crystalline regions in starch granules [su-genotype  among investigated maize starches  are expressed in the form of disturbances in the starch polysaccharide branching process, as opposed to the other tested starches from the wx and “ae” genotypes. This fact, however, requires further and more profound research.It is worth mentioning that the thermodynamic melting parameters of the enotypes . Simultaomplexes . SpecifiAs known, the melting of polymer crystals begins with the melting of defects in an amorphous lamella. Here, amylopectin B-chains and amylose “tie chains” located in amorphous lamellae defects play the role of such defects ,44. The 0m (366.5 K) and ΔH (35.5 J/g) as well as pcrl (1.48 g/cm3) for the A-type crystalline spherulites to which maize starches are attributed [ae” genotypes compared to the corresponding value for starches from wx-genotype. This phenomenon indicated that defects in the starch structure accumulated with increasing amylose content in maize starch of the transition from wx to “ae”-genotype. It is clear from the presented data (su-genotypes (Alina and Rannaya Lacomka varieties), probably due to their less-ordered structure compared to other starches studied. Note that for starches of the “ae” genotype with an amylose content of 32.0% and 38.0%, the value of the entropy of the end faces was close to the corresponding value for starches of the WT genotype with an amylose content of 15% and the su-genotype of the Baksanskaya sakharnaya variety, which probably indicated about the similarity in the degree of their order. It should be noted that starches with ploidy equal to four (varieties Populyatsiya MRPP22 and Baksanskaya sakharnaya) were characterized by closely free surface entropy values. It is likely that these starches were characterized by the same number of less-ordered structures. It can be assumed that the number of genes of diploid and tetraploid genotypes also determines the thermodynamic and structural features of starches, but this fact requires further research.The accumulation of defects accompanies the development of crystallites with a more ‘mellow’ surface. Thermodynamic parameters were assessed for the surface of the faces of crystalline lamellae in the tested starches . The caltributed . The valted data  that theae” genotypes. The amylose content and the genotype of plants used for extracting starches determined changes in the thermodynamic parameters of investigated maize starches.Thus, an increase in the amylose content in the tested maize starches accompanied a decrease in their thermodynamic melting parameters; simultaneously, the amount of amylose–lipid complexes remained practically unchanged for starches from WT and “wx, “ae”, su, and transitional from wx to “ae” starch genotypes. The samples of maize varieties Baksanskaya sugar (su-genotype) and Population MRPP22  had a tetraploid genotype but did not show signs of a pronounced specific effect of genome ploidy on the composition or ratio of amylose/amylopectin.The studied high-starch maize accessions contained different starch genotypes with variable amylose/amylopectin ratios. Among the starch types of maize studied, four groups comprised the ae” genotype had the highest values in granulometric sizes and from su-genotype—the minimum granulometric sizes among the tested starches. An increase in the amylose content in investigated maize starches with different genotypes accompanied a decrease in their thermodynamic melting parameters and the accumulation of defective structures in the studied starches, regardless of their ploidy. The starches extracted from the su-genotype differed in their thermodynamic melting parameters from other accessions having lower crystallinity and the highest content of less-ordered structures. Thus, amylose content in starch and the individual features of the maize genotype determine the thermodynamic melting parameters of studied starches. Probably the ploidy of maize determines the physicochemical properties of the extracted starches, but this requires further research.As shown, starch granules of the “wx genotype , with wild genotype maize accessions (varieties Luch and Kabardinskaya belaya zubovidnaya), and accession with the su genotype  are of practical importance: they may serve as valuable raw materials for starch and treacle production industries and as sources for the development of new high-starch cultivars of waxy maize and traditional grain cultivars with a high mass fraction of starch. The studied high-starch waxy maize accessions with the su gene on the phenotypic, morphological, and thermodynamic characteristics of native maize starch. Our study shows that genes involved in maize starch biosynthesis have a multifactorial effect, which manifests itself as a regulation of the parameters of thermodynamic and structural features of maize starch and biochemical ones of grain. These features may change more under the influence of several different in their effects genes than their number. Breeding and genetic engineering techniques make it possible to obtain starch with a wide range of physicochemical properties and starch grain phenotypes due to various combinations of several genes  in the genome of a diploid or tetraploid maize plant affecting starch synthesis. The study of the features of various maize genotypes, deciphering their genetic components, and their effects affecting the properties of starch will significantly improve the technology for producing cereal starches with specified physical and chemical properties as raw materials for the relevant industries.A comparative analysis of tetraploid and diploid samples differing only in the presence of one allele of the sugar content gene in the dominant and recessive states clearly showed a significant effect of the


Vascular aging in large arteries plays an important role in contributing to cardiovascular morbidity and mortality . The subWhile the thickening of the arterial walls should be much more correlated with atherosclerotic processes, arterial stiffening aetiology should be linked to degenerative/calcified processes. However, the mechanisms through which both of these processes are related have yet to be clarified. Recently, the Consensus Document  on ventrIn the recent literature, there has been emerging interest in the influence of the parameters that reflect the aortic stiffness of patients suffering with HF. In research on the possible effect of PWV on HF , we docup = 0.006, p < 0.001, and p < 0.0001, respectively). In particular, a PWV pre-discharge value of ≥ 10 m/s was inversely correlated with free-event survival . These data suggest that non-invasive measurements of arterial stiffness in HF patients discharged alive after an acute decompensation might be considered as useful prognostic parameters [Moreover, the values of augmentation index AIx75) in the HF group vs. the CVRF and healthy population seemed to be different , demonstrating significantly lower values for the HF group. It can be hypothesized that a reduction/delay in the arrival of the reflected wave can enter into the determinism of the left ventricular remodeling, since the arrival of the reflected wave creates an additional obstacle to the ejection of the left ventricle. Furthermore, the statistical analysis demonstrated that the correlation between the values of PWV/AIx75 and the value of the renal filtrate (GFR) was significantly the strongest, as underlying renal dysfunction might determine an important effect that increases the aortic stiffness, which could be crucial in the evolution of both heart failure and vascular damage. More recently [ in the Hp = 0.03), together with a reduction in their PWVs from 13.7 ± 2.4 m/s to 10.5 ± 1.43 m/s (p < 0.05) [p < 0.05) [Alirocumab, a human monoclonal antibody to proprotein convertase subtilisin–kexin type 9 (PCSK9), improved the cardiovascular outcomes after an acute coronary syndrome in patients receiving high-intensity statin therapy and clearly reduced the plasma level of their LDL cholesterol after a 24-month treatment . Upon tr < 0.05) . A lineaIn conclusion, the analysis of arterial stiffness seemed to be promising for the determinism and prognosis of HF patients. In the search for a prognostic meaning in HF patients, aortic stiffness seems to be particularly related to renal dysfunction and is extremely important in the prognostic setting. Finally, the huge reduction in LDL cholesterol obtained with PCSK9 inhibitors might influence the arterial stiffness of treated patients, even in a short/mid-term follow-up.


There is an unmet need for optimizing hepatic allograft allocation from nondirected living liver donors (ND-LLD).Using OPTN living donor liver transplant (LDLT) data (1/1/2000-12/31/2019), we identified 6328 LDLTs . Random forest survival models were constructed to predict 10-year graft survival for each of the 3 graft types.Donor-to-recipient body surface area ratio was an important predictor in all 3 models. Other predictors in all 3 models were: malignant diagnosis, medical location at LDLT (inpatient/ICU), and moderate ascites. Biliary atresia was important in left and left-lateral graft models. Re-transplant was important in right graft models. C-index for 10-year graft survival predictions for the 3 models were: 0.70 ; 0.63 (left); 0.61 (right). Similar C-indices were found for 1-, 3-, and 5-year graft survivals. Comparison of model predictions to actual 10-year graft survivals demonstrated that the predicted upper quartile survival group in each model had significantly better actual 10-year graft survival compared to the lower quartiles (p<0.005).When applied in clinical context, our models assist with the identification and stratification of potential recipients for hepatic grafts from ND-LLD based on predicted graft survivals, while accounting for complex donor-recipient interactions. These analyses highlight the unmet need for granular data collection and machine learning modeling to identify potential recipients who have the best predicted transplant outcomes with ND-LLD grafts. In the past decade, the number of living donor liver transplants (LDLTs) in the United States, along with the number of transplant centers performing LDLTs, have both increased –3. Over Living donors interested in nondirected liver donation frequently express desire for their donation to affect the greatest good from the graft. ND-LLD grafts offer transplant centers the opportunity to help patients who may be underserved by the current deceased donor Model for End-Stage Liver Disease (MELD)-based allocation system, for example, individuals with small stature or low MELD patients with portal hypertensive complications. Additionally, ND-LLDs enable liver transplant centers to consider more novel undertakings such as living liver donor chains. Despite the recognized importance of ND-LLDs, there is no standardized evidence-based approach to help guide liver transplant centers in ND-LLD graft allocation. Currently, individual centers develop their own internal preferences and policies regarding how to internally allocate liver grafts from ND-LLDs, with all U.S. programs abiding by the the National Organ Transplant Act that was implemented in 1984, making it illegal to buy or sell human organs and tissues in the U.S. United Network for Organ Sharing (UNOS) policy 14.6.B allows for utility-based allocation processes, and guidance from the Organ Procurement and Transplantation Network (OPTN) recommends that liver transplant centers ‘make an effort to match donors and candidates appropriately’ and ‘maximize the potential good’ that will emerge from ND-LLD grafts . We therWe conducted a retrospective analysis of the OPTN liver dataset. The dataset contained information on U.S. recipients who underwent liver transplant from living liver donor grafts from 1/1/2000 to 12/31/2019, with follow-up to 3/20/2020. Data from both directed and nondirected living liver donation was collected. Donor and recipient data were recorded from the transplant candidate and transplant recipient forms. We excluded recipients of deceased donor grafts, whole liver grafts, and multiorgan transplants.The donor data collected included: age at donation, sex, race, history of cigarette smoking, ABO blood type, height (cm), weight (kg), and calculated donor body surface area  using tRecorded recipient factors included: age at time of transplant , sex, height (cm), weight (kg), calculation of recipient BSA, etiology of underlying liver disease, calculated MELD score or Pediatric End-Stage Liver Disease (PELD) score at transplant, serum albumin level at transplant, presence of ascites , encephalopathy , presence of portal vein thrombosis, previous abdominal surgery, transjugular intrahepatic portosystemic shunt, diabetes mellitus diagnosis, location at time of transplant , need for mechanical ventilation and/or pressor support, on dialysis the week prior to transplant, Epstein-Barr virus serostatus, and Cytomegalovirus serostatus.The dataset used to develop the analytic models described in this study is available through the OPTN.All data obtained from human participants were obtained in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.The interpretation and reporting of these OPTN data are the responsibility of the authors and in no way should be considered an official policy of, or interpretation by, the OPTN or the U.S. Government. The University of Washington Human Subjects Division deems that the OPTN database is de-identified and publicly available, and thus this study was exempt from human subjects review.Continuous variables were depicted as median and interquartile ranges (IQR). Categorical variables were presented as percentages. For all data, if <1% of the categorical values were missing, the majority value was given. For the 305 donors and 66 recipients with missing body weight, values were imputed with linear regression using age, sex, and race. For the 66 donors and 16 recipients with missing height, values were imputed with linear regression using age, sex, and race. For the 111 recipients missing albumin level, the median albumin value of 3.1 was given. For 107 recipients with missing values for ascites, the value of ‘absent’ was given. Sensitivity analyses revealed no change in the final results by imputing any of the values. Chi-square analysis was used to compare categorical variables. One-way ANOVA was used to compare continuous variables. Donor-to-recipient BSA ratio (D-R-BSA) was calculated for each donor-recipient pair. Determining the critical ratio (or cut-off) for D-R-BSA for each graft was performed using multiple methods . MultiplMachine learning survival models are increasingly being utilized in the field of transplantation . These mp-value <0.05. We also modeled 5-year, 3-year- and 1-year graft survivals, and found similar c-indices as for 10-year graft survival (data not shown). The Chi-square, ANOVA, Kaplan-Meier, and Cox proportional hazard analyses were performed using JMP-Pro Version 15.1.0 . The RFS analyses were performed using R version 4.0.0. and the randomForestSRC and the random package , which is the proportion of observations that the statistical model can order correctly in terms of survival times and, for binary outcomes, may be considered conceptually similar to the area under the receiver operating characteristic curve (AUC) , 18. Due package .p<0.001) between all graft types, with D-R-BSA ratios (median and IQR) of 4.3 (3.1-5.3), 1.21 (1.1-1.5), and 1.0 (0.9-1.1) for the left-lateral, left, and right grafts, respectively. Other donor and recipient characteristic comparisons by graft type  are provided in We included data on all 6328 living donor liver transplant recipients from 1/1/2000 to 12/31/2019. LDLT from non-directed donors accounted for 5% of total LDLDs during the study period. There were 1063 left-lateral, 644 left, and 4621 right grafts. Of the left-lateral grafts, 1001 (94%) were transplanted into recipients aged 0-12 years, and 62 (6%) transplanted into older recipients (> 12 years). Of the left grafts, 213 (33.0%) were transplanted into recipients aged 0-17 years, while 431 (67%) were transplanted into older recipients (>17 years). Of the right grafts, 100 (2%) were transplanted into recipients aged 0-17 years, compared to 4521 (98%) transplanted into older recipients (>17 years). More females received left grafts (61%), whereas more males (58%) received right grafts. Patients with cholestatic liver diseases (primary biliary cirrhosis [PBC] and primary sclerosing cholangitis [PSC]) accounted for 24% of left and 24% of right graft recipients. The D-R-BSA ratio was significantly different (p<0.001) between graft types, with 194 (18%) losses for the left-lateral graft, compared to 188 (29%) and 1308 (28%) for the left and right grafts, respectively. There was no difference in graft survival when comparing left to right grafts. The Kaplan-Meier graft survival curves confirming the importance of the critical ratio for D-R-BSA  for each graft type are shown in p-value 0.002 for left-lateral, p=0.001 for left, p>0.001 for right).The Kaplan-Meier graft survival curves comparing the three graft types are shown in The importance of the selected recipient and donor-to-recipient variables using random forest survival models is demonstrated graphically by LIME plots in p<0.05), indicating that the model predictions have good discrimination for identifying which recipients will maximize long-term graft survival.To compare model predictions to actual graft survival, the models’ 10-year graft survival predictions were divided into groups consisting of the upper quartile and the lower three quartiles. The Kaplan-Meier actual graft survival curves for each model prediction group (upper quartile vs lower quartiles) are shown in At the University of Washington, we began using this utility-based model in 2020 to assist our team in making evidence-based, less subjective decisions regarding allocation of hepatic allografts from nondirected living liver donors. Our process is not intended to be based solely upon the modeling results, making the actual values of the modeling c-indices less intrinsic to the selection process. Our approach to non-directed donor allocation incorporates our objective modeling results into our teams’ clinical assessments of potential recipients. Therefore, our decision-making process proceeds as follows: (1) using our liver transplant program’s list of patients who are active on the UNOS waiting list, we run our utility-based allocation model for the nondirected donor, taking into the lobe of liver being donated  and accounting for blood type compatibility between the nondirected donor and potential recipients. Additionally, our transplant surgeon specifies body size parameters for potential recipients  based upon the donor’s graft volume; (2) the list of potential recipients generated from step 1 is rank ordered (descending) according to predicted probability of 10-year graft survival, with 65% as our lower limit cutoff for 10-year graft survival; (3) separately, a match run using our nondirected donor is conducted; (4) the rank ordered list generated from our utility model in step 2 is reviewed in a thoughtful multi-disciplinary clinical forum attended by our transplant hepatologists and transplant surgeons to review each individual potential recipient’s clinical indications for liver transplant, portal hypertensive complications, and surgical complexity that might impact suitability for receiving a living donor hepatic allograft. When there is more than one suitable potential recipient for the nondirected living liver donor graft, then the match run sequence is used to prioritize the potential recipients (a lower match sequence number confers higher priority).To further characterize the potential recipients identified by our utility models, we compared these individuals to the potential recipients identified on the match run list. In this comparison, we found that potential recipients identified via the utility model had lower match MELD scores , lower BMIs , and similar Donor-to-Recipient BSA ratios . There were no significant differences in recipient characteristics between the utility model and match run with regards to: age, height, weight, sex, cholestatic liver disease diagnosis, or diagnosis of HCC (data not shown). Our nondirected donors have all been right-lobe donors thus far, but a similar process to that outlined above could be implemented for left or left lateral segment donors.Nondirected  living liver donors present an important opportunity for expanding the donor organ pool. Currently, there is an array of terminology used in the living liver donor literature to describe these donors; therefore, to avoid confusion in this study, we have coined the term nondirected living liver donor (ND-LLD) to clearly represent the donor population to which we are referring. We present an original, proof of concept, evidence-based model to help identify and stratify potential recipients of ND-LLD livers, based on predicted 10-year graft survival, while accounting for complex donor-recipient interactions. We propose that this model is best applied to help guide thoughtful clinical decision-making, rather than advocating that any modeling results be used in isolation. Our model best predicted 10-year graft survival for left-lateral grafts (c-index 0.7) followed by left grafts (c-index 0.63) and right grafts (c-index 0.61). Model predictions correlated with actual recipient survival, indicating good discrimination for predicting which recipients will maximize liver graft survival. Our models are useful for objectively guiding ND-LLD graft allocation in the clinical setting. Our clinical liver transplant team has found this allocation process to be very efficient and beneficial in ensuring that we are evidence-based in the selection of a potential recipient, aiming to maximize the utility of the generous gift of the living liver donor’s graft. Additionally, our objective allocation process allows us to be transparent with our non-directed living liver donors when we discuss how the graft will be allocated, fostering increased trust between our transplant program and the donor. While we chose our desired outcome measure to be 10-year graft survival in an effort to maximize utility of the graft, we also investigated 5-, 3- and 1-year graft survivals and found similar modeling performances as in our 10-year outcomes models, so only our 10-year data are presented in detail in this manuscript.To our knowledge, this is the first model using machine learning techniques to incorporate objective donor, recipient, and combined donor-to-recipient characteristics available in the OPTN data to predict long-term graft survival after LDLT. We present the results from our RFS models, though we did evaluate several other machine-based learning models (data not shown). The RFS models were the best models for the UNOS dataset and the models with the best validation.Importantly, our models incorporate donor-to-recipient BSA ratios, which estimate liver volume more accurately than donor and recipient body weight and height alone , 10, 20.The use of RFS models also allowed us to consider many donor and recipient characteristics, as well as the complex interactions between them, in our algorithm. The RFS model was chosen over the Cox proportional hazard model to improve the generalizability and accuracy of the model, given the complexity of the data, and expected interactions between terms, and the potential implications these interactions can have on candidate selection for liver transplantation. The Cox proportional hazard model has difficulty with interactions between terms, making its models less interpretable and accurate . We inclOur model adds to the existing literature that supports the use of artificial intelligence (AI) and machine learning models to optimize liver transplant care. Studies have already shown that AI models can improve the accuracy of predicting waitlist mortality  and postOur study has several strengths. First, we utilized national LDLT data from the U.S. OPTN database to inform our analytic models, including adult-to-adult and adult-to-child transplants, which allows for greater generalizability of our findings. Second, in addition to developing graft survival prediction models, we also confirmed the models’ abilities to discriminate survival by comparing the accuracy of predictions to actual graft survival. Third, the use of RFS models allowed us to account for a large number of donor and recipient variables, along with their potential interactions.We acknowledge the limitations of this study. Although our models were developed using a large, national dataset, this was a retrospective analysis. In addition, the c-index for our left and right models were <0.7, suggesting that other factors that are not currently included in the U.S. OPTN data, may improve the fit of the models , 36. ForFinally, within the context of the national data used for these analyses, we could not account for individual transplant center experience, such as LDLT volume, frequency of LDLT surgery, or surgical experience, which have been shown to impact LDLT outcomes , 37, 38.In conclusion, there is an unmet need for more evidence to help guide living liver donor programs in the allocation of hepatic grafts from nondirected ‘anonymous’ living liver donors in the U.S. In this original, proof of concept study, our models present real world evidence from real world data to help address this unmet need, and include complex donor-recipient interactions to accurately predict 10-year graft survival for left-lateral, left, and right grafts. The model was particularly robust for left-lateral grafts. Therefore, our results demonstrate that machine learning is useful in the development and implementation of allocation models that can guide transplant clinical decision making and help optimize outcomes for recipients of ND-LLD hepatic grafts. As the opportunities for LDLT and ND-LLD donation expand, our findings set the stage for further development of robust allocation strategies for ND-LLD grafts.https://www.srtr.org/about-the-data/the-srtr-database/.Publicly available datasets were analyzed in this study. This data can be found here: Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. Written informed consent from the participants’ legal guardian/next of kin was not required to participate in this study in accordance with the national legislation and the institutional requirements.KB, SB, JP, and MS contributed to the design and implementation of the research. JP, KB, and SB contributed to the analyses of the results. NK, KB, SB, JP, CK, MS, JR, PH, RB, and AD participated in critical data interpretation and writing of the manuscript. KB, SB, MS, and JP conceived the original idea and supervised the project. All authors contributed to the article and approved the submitted version.


Aims: To synthesize and evaluate the available scientific evidence on the efficacy of antihypertensive drugs on arterial stiffness in patients with hypertension by using a network meta-analysis approach.Methods: A systematic search of the MEDLINE (via PubMed), Scopus, and Web of Science databases was conducted to identify experimental studies addressing the effect of different antihypertensive drugs on arterial stiffness parameters (pulse wave velocity [PWV] and augmentation index [AIx]) in adults with hypertension. Comparative evaluation of the effect of antihypertensive drugs was performed by conducting a standard pairwise meta-analysis and a network meta-analysis for direct and indirect comparisons between antihypertensive drugs and placebo/other antihypertensive drugs. Analyses were performed including studies of any duration and only studies longer than 6 months length.Results: Seventy-six studies were included in the main analysis and considering only studies longer than 6 months length, thiazide diuretics, ACEIs, ARBs, the ACEI/ARB combination, the ACEI/CCB combination, and the ARB/CCB combination showed a higher effect on reducing PWV, and ACEIs and ARBs on reducing AIx.Conclusion: Our research provides evidence that antihypertensive medications are an effective way to treat arterial stiffness in adults with hypertension. Based on our findings, patients with hypertension who have greater levels of arterial stiffness may benefit from using thiazide diuretics, ACEIs, ARBs, the ACEI/ARB combination, the ACEI/CCB combination, and the ARB/CCB combination.Systematic Review Registration: PROSPERO (CRD42021276360). Increased blood pressure is one of the major risk factors for cardiovascular disease, affecting more than one-third of the population, accounting for 20.5% of the global burden of cardiovascular disease and the leading cause of death and disability worldwide. . Large aper se rather than to specific drug properties.   and the ClinicalTrials.gov) and on drug approval agency websites. Prior to the final analyses, the searches were repeated only to include all current and potential studies.The literature search was conducted through the MEDLINE, Scopus, Cochrane Central Register of Controlled Trials, Cochrane Database of Systematic Reviews, and Web of Science databases. The above searches were supplemented by manual searches of published or ongoing randomized controlled trials (RCTs) in international trial registries  (>18 years of age and of both genders) were selected. If two or more studies provided data on the same sample, the one that presented the most detailed results or provided the largest sample size was chosen. Types of Intervention: Studies using any of the different drugs in the antihypertensive groups as an intervention , as wellIndeed, since PWV  is a measure of large artery stiffness, whereas AIx is an integrated measure of arterial stiffness and wave reflection, these two measures are analysed separately.Based on the recommendations of the Cochrane Collaboration Handbook, two authors independently conducted the risk of bias assessment. . DisagreThe risk of bias of RCTs was assessed using the Cochrane Collaboration’s tool for assessing risk of bias (RoB2). .We used the Grading of Recommendations, Assessment, Development and Evaluation (GRADE) tool to assess the evidence quality and provide recommendations. .ad hoc table describing direct and indirect comparisons.We qualitatively summarize the included RCTs in an The present NMA was conducted as follows: we assessed the strength of the available evidence using a network geometry graph to display the evidence in the network for arterial stiffness (PWV and AIx). In addition, the network geometry graph to show the evidence in the network for arterial stiffness (PWV and AIx) was performed including only studies longer than 6 months length. In this graph, the size of the nodes was proportional to the number of participants in trials who received the intervention specified in the node, and the thickness of the continuous line connecting nodes was proportional to the number of trials directly comparing the two treatments. .d values was calculated, as an estimate of effect size (ES). In addition, these analyses were performed by including only studies longer than 6 months length. We assessed heterogeneity using the I2 statistic, , moderate (30%–60%), substantial (60%–75%), or considerable (75%–100%). We also considered the corresponding p values. Furthermore, the size and clinical relevance of heterogeneity was determined by the τ2 statistic.Comparative evaluation of the intervention effect on arterial stiffness (PWV and AIx) was performed by conducting a random effects pairwise meta-analysis and a frequentist NMA for comparisons between interventions and controls. Cohen atistic, , rangingSensitivity analysis was conducted to evaluate the robustness of the pooled estimates, and a reanalysis was conducted by eliminating one study at a time.versus patients with hypertension and other pathologies).Subgroup analyses were conducted based on the type of PWV  and on the type of population .The probability that each intervention is the most effective was presented by rankograms. In addition, for each intervention, we estimated the surface under cumulative ranking (SUCRA). . With SUPublication bias was tested using Egger’s regression asymmetry test, , settingThe analyses were performed using STATA 15 .Seventy-six studies  addressiIn the included populations, a total of 5413 patients with arterial hypertension were aged between 37.0 and 72.8 years. The duration of treatment with antihypertensive drugs in the studies ranged from 4 to 208 weeks.As evaluated by the RoB2 tool, 80.3% of the studies showed some concerns in the risk of bias, with 10.5% and 9.2% studies showing high and low risks of bias in the overall bias, respectively. .When the quality grading of evidence of each pairwise comparison was evaluated for PWV using the GRADE system, 3.1% of the pairwise comparisons were categorized as high, 45.2% as moderate, 31.3% as low and 19.4% as very low . When thNetwork available comparisons between different types of antihypertensive drugs on PWV are shown in Furthermore, network available comparisons between different types of antihypertensive drugs on AIx are shown in For PWV, the ACEI/ARB combination showed the highest SUCRA (93.0%) . For AIxBased on the type of PWV, beta-blockers , ACEI , ARB , the ACEI/ARB combination , the ACEI/CCB combination , the ARB/CCB combination , and the thiazide diuretics/ARB combination  were effective in reducing central PWV , and ACEBased on the type of population, the ACEI/ARB combination  and ACEI  were effective in reducing PWV , and ACEversus CCB and diastolic blood pressure reduction in ACEI versus CCB were related to pooled ES estimate , ACEI versus ARB for PWV and AIx , ARB versus the ACEI/ARB combination for PWV , ARB versus placebo for AIx , beta-blockers versus ACEI for AIx , and diuretics versus ARB for AIx  (Considerable heterogeneity was found for ACEI  = 1.38) .versus CCB (p = 0.005), diuretics versus CCB (p = 0.041) and the thiazide diuretic/ACEI combination versus the ACEI/CCB combination (p = 0.040) for PWV and for ACEI versus ARB (p = 0.039) for AIx.Finally, there was evidence of a small study effect in Egger’s test for ARB This NMA provides an overview of the evidence comparing the effect of antihypertensive drugs on reducing arterial stiffness in patients with hypertension. Although all types of hypertensive drugs reduced arterial stiffness measured by PWV and AIx, beta-blockers, ACEI, ARB, the ACEI/ARB combination, and the ARB/CCB combination showed a higher effect on reducing PWV, and ACEIs, ARBs, CCBs, renin inhibitors, the thiazide diuretic/ACEI combination and the ARB/CCB combination on reducing AIx. When only studies longer than 6 months length were included, thiazide diuretics, ACEI, ARB, the ACEI/ARB combination, the ACEI/CCB combination, and the ARB/CCB combination showed a higher effect on reducing PWV, and ACEIs and ARBs on reducing AIx. Additionally, when analyses were performed in patients with hypertension only (without other comorbidities), the ACEI/ARB combination and ACEI showed a higher effect on reducing PWV and AIx, respectively. Finally, beta-blockers, ACEI, ARB, the ACEI/ARB combination, the ACEI/CCB combination, the ARB/CCB combination, and the thiazide diuretics/ARB combination showed a statistically significant effect on reducing central PWV, and the ACEI/ARB combination on reducing peripheral PWV. However, it is worth noting that the ACEI/ARB combination of, despite showing improvement in stiffness measures, has been associated with an increased risk of cardiovascular events. As such, caution should be exercised when considering the use of this combination therapy in hypertensive patients, and alternative approaches may be warranted to mitigate potential risks. .versus placebo. They found that PWV decreased between −0.75 and −1.3 m/s in the treatment group compared with placebo, which decreased between −0.17 and −0.44 m/s. In short-term trials, ACEIs were more effective than CCBs, and in long-term trials, ACEIs, CCBs, beta-blockers and diuretics were more effective than placebo. The meta-analysis by Chen et al. (versus other antihypertensive agents (except ACEIs) in reducing PWV and AIx. ARBs were not found to be superior to other types of antihypertensive agents in lowering PWV, but the ability of ARBs to improve Aix was superior. Another meta-analysis published by Shahin et al. . These authors concluded that ACEIs reduce PWV and Aix. However, due to the lack of high-quality and adequately powered RCTs, it was unclear whether the effect of ACEIs on arterial stiffness was superior to that of other antihypertensives. In the meta-analysis by versus placebo showed statistically significant differences. In the same vein, longitudinal studies that have analysed the effect of ACEIs or ARBs alone, in combination or in combination with CCBs or diuretics have been shown to be effective in reducing arterial stiffness, , ACEIs (high decrease for PWV and AIx), ARBs (high decrease for PWV and moderate decrease for AIx) and CCBs (moderate decrease for AIx) are the pharmacological groups with the greatest effect in reducing arterial stiffness. The results found in this NMA on the effect of different types of antihypertensive drugs on arterial stiffness are consistent with data published in previous meta-analyses. Thus, the meta-analysis by Ong et al.  includedn et al.  includedn et al.  analysediffness, , althougiffness, .Thus, all results suggest that antihypertensive agents may have beneficial effects on arterial stiffness and central hemodynamic parameters, but the effect on arterial stiffness differs between them. Possible explanations for these differences include that ACEIs and ARBs influence arterial stiffness by reducing fibrosis and increasing arterial wall distensibility. In addition, some authors have suggested other mechanisms, such as reduction of oxidative stress, inflammation, and vasodilation through inhibition of angiotensin II, favoring vascular remodelling and endothelial function. . The mecSimilarly, the effect varies according to the type of measure used to assess arterial stiffness, and we found less effect on PWV than on AIx. AIx is an index of wave reflection, which is influenced not only by arterial stiffness but also by microcirculatory status and cardiac contractility, which are highly dynamic, more than structural parameters such as arterial stiffness. .Although the blood pressure response to different antihypertensive drug classes may differ according to the age of the patient, , we did The main limitations of this NMA are as follows: Some drug combinations in the treatment of hypertension, such as ACEIs and ARBs, have been included, , as wellTherefore, it is necessary to plan future RCTs with the 5 classes of drugs currently indicated for the treatment of hypertension: : ACEIs, In summary, our study supports that antihypertensive drugs are a suitable treatment to reduce arterial stiffness in patients with hypertension. Based on our results, ACEIs, ARBs, beta-blockers, CCBs, renin inhibitors, the thiazide diuretics/ACEI combination, the ARB/CCB combination and the ACEI/ARB combination could be useful for patients with hypertension who have higher levels of arterial stiffness. Additionally, if we only consider antihypertensive treatments longer than 6 months, thiazide diuretics, ACEI, ARB, the ACEI/ARB combination, the ACEI/CCB combination, and the ARB/CCB combination are the most effective treatments. This result is clinically relevant since arterial stiffness is a closely related factor to hypertension, which produces a global burden of cardiovascular disease and is the leading cause of death and disability worldwide. However, it is worth noting that the ACEI/ARB combination, despite showing improvement in stiffness measures, has been associated with an increased risk of cardiovascular events. As such, caution should be exercised when considering the use of this combination therapy in hypertensive patients, and alternative approaches may be warranted to mitigate potential risks. Notwithstanding, it is essential that future well-designed, statistically powered RCTs are conducted to strengthen the currently weak evidence.