Stored platelet hemostatic phenotype and function is not altered when donors are on testosterone replacement therapy.

BACKGROUND: Critical shortages in the national blood supply have led to a re-evaluation of previously overlooked donor sources for blood products. As a part of that effort, red blood cells collected from therapeutic phlebotomy of donors on testosterone replacement therapy (TRT) have been conditionally approved for transfusion. However, platelets from TRT donors are not currently approved for use due to limited data on effects of supraphysiologic testosterone on recipient safety and platelet function. The objective of this study was to provide a comprehensive profile of phenotype and function in platelets from TRT and control donors. STUDY DESIGN AND METHODS: Platelets in plasma were collected from TRT and control donors (N = 10 per group; age- and sex-matched) and stored at room temperature for 7 days. On storage Day 1 (D1) and Day 7 (D7), platelet products were analyzed for platelet count, metabolic parameters (i.e., glucose, lactate, mitochondrial function), surface receptor expression, aggregation, thrombin generation, and thrombus formation under physiological flow conditions. RESULTS: TRT donor platelets were not significantly different than control donor platelets in terms of count, surface phenotype, metabolic function, ability to aggregate, thrombin generation, or ability to form occlusive thrombus under arterial flow regimes. Both groups were similar to each other by D7, but had significantly lost hemostatic function compared to D1. DISCUSSION: Platelets derived from donors undergoing TRT have similar phenotypic and functional profiles compared to those derived from control donors. This suggests that therapeutic phlebotomy of TRT donors may provide a useful source for platelet products.

Testosterone supplementation increases red blood cell susceptibility to oxidative stress, decreases membrane deformability, and decreases survival after cold storage and transfusion.

BACKGROUND: Blood collection from donors on testosterone therapy (TT) is restricted to red blood cell (RBC) concentrates to avoid patient exposure to supraphysiological testosterone (T). The objective of this study was to identify TT-related changes in RBC characteristics relevant to transfusion effectiveness in patients. STUDY DESIGN: This was a two-part study with cohorts of patients and blood donors on TT. In part 1, we conducted longitudinal evaluation of RBCs collected before and at three time points after initiation of T. RBC assays included storage and oxidative hemolysis, membrane deformability (elongation index), and oximetry. In part 2, we evaluated the fate of transfused RBCs from TT donors in immunodeficient mice and by retrospective analyses of NIH's vein-to-vein databases. RESULTS: TT increased oxidative hemolysis (1.45-fold change) and decreased RBC membrane deformability. Plasma free testosterone was positively correlated with oxidative hemolysis (r = .552) and negatively correlated with the elongation index (r = -.472). Stored and gamma-irradiated RBCs from TT donors had lower posttransfusion recovery in mice compared to controls (41.6 ± 12 vs. 55.3 ± 20.5%). Recipients of RBCs from male donors taking T had 25% lower hemoglobin increments compared to recipients of RBCs from non-TT male donors, and had increased incidence (OR, 1.80) of requiring additional RBC transfusions within 48 h of the index transfusion event. CONCLUSIONS: TT is associated with altered RBC characteristics and transfusion effectiveness. These results suggest that clinical utilization of TT RBCs may be less effective in recipients who benefit from longer RBC survival, such as chronically transfused patients.

Genetic determinants of plasma testosterone in male blood donors are associated with altered red blood cell characteristics and survival in cold storage and after transfusion.

Genetic mutations in genes regulating plasma testosterone in men may interfere with effective erythropoiesis, and may result in red blood cell (RBC) dysfunction and hemolysis. The aim of this study was to identify genetic polymorphisms in male donors that regulate plasma testosterone and impact RBC survival in cold storage and after transfusion. We evaluated nine single nucleotide polymorphisms (SNPs) previously reported to be associated with circulating testosterone in male plasma. These SNPs were linked with donor-component-recipient databases (NIH REDS program) to determine SNP associations with donor RBC hematological indices, osmotic and oxidative hemolysis, and RBC transfusion effectiveness defined as adjusted hemoglobin increments (delta hemoglobin, ΔHb) following a single RBC unit transfusion. Four of the nine testosterone SNPs were located on the X chromosome, of which two (rs7057002, rs73629199) were significantly associated with reduced hemoglobin increments (0.2 and 0.3 g/dL, respectively) compared with reference alleles in transfused recipients. Seven of the nine testosterone SNPs were associated with significant changes in RBC susceptibility to osmotic hemolysis including a missense mutation in the major plasma carrier of testosterone (SHBG, rs6259), and four SNPs with changes in oxidative hemolysis. Four SNPs were associated with decreased RBC count, hemoglobin, and hematocrit. Ancestry/ethnicity-specific (African and Hispanic) associations were observed between two SNPs (rs7057002, rs7879462) and oxidative hemolysis. Genetic determinants of plasma testosterone in male donors significantly impact the quality and transfusion effectiveness of cold stored RBCs. Testosterone SNPs associated with decreased RBC transfusion effectiveness may have clinical implications and warrant further revaluation.

Dry fractionation process operations in the production of protein concentrates: A review.

The market for plant proteins is expanding rapidly as the negative impacts of animal agriculture on the environment and resources become more evident. Plant proteins offer competitive advantages in production costs, energy requirements, and sustainability. Conventional plant-protein extraction is water and chemical-intensive, posing environmental concerns. Dry fractionation is an energy-efficient and environmentally friendly process for protein separation, preserving protein's native functionality. Cereals and pulses are excellent sources of plant proteins as they are widely grown worldwide. This paper provides a comprehensive review of the dry fractionation process utilized for different seeds to obtain protein-rich fractions with high purity and functionality. Pretreatments, such as dehulling and defatting, are known to enhance the protein separation efficiency. Factors, such as milling speed, mill classifier speed, feed rate, seed type, and hardness, were crucial for obtaining parent flour of desired particle size distribution during milling. The air classification or electrostatic separation settings are crucial in determining the quality of the separated protein. The cut point in air classification is targeted based on the starch granule size of the seed material. Optimization of these operations, applied to different pulses and seeds, led to higher yields of proteins with higher purity. Dual techniques, such as air classification and electrostatic separation, enhance protein purity. The yield of the protein concentrates can be increased by recycling the coarse fractions. Further research is necessary to improve the quality, purity, and yield of protein concentrates to enable more efficient use of plant proteins to meet global protein demands.

Genetic control of source-sink relationships in grain sorghum.

MAIN CONCLUSION: QTL hotspots identified for selected source-sink-related traits provide the opportunity for pyramiding favorable alleles for improving sorghum productivity under diverse environments. A sorghum bi-parental mapping population was evaluated under six different environments at Hays and Manhattan, Kansas, USA, in 2016 and 2017, to identify genomic regions controlling source-sink relationships. The population consisted of 210 recombinant inbred lines developed from US elite post-flowering drought susceptible (RTx430) and a known post-flowering drought tolerant cultivar (SC35). Selected physiological traits related to source (effective quantum yield of photosystem II and chlorophyll index), sink (grain yield per panicle) and panicle neck diameter were recorded during grain filling. The results showed strong phenotypic and genotypic association between panicle neck diameter and grain yield per panicle during mid-grain filling and at maturity. Multiple QTL model revealed 5-12 including 2-5 major QTL for each trait. Among them 3, 7 and 8 QTL for quantum yield, panicle neck diameter and chlorophyll index, respectively, have not been identified previously in sorghum. Phenotypic variation explained by QTL identified across target traits ranged between 5.5 and 25.4%. Panicle neck diameter and grain yield per panicle were positively associated, indicating the possibility of targeting common co-localized QTL to improve both traits simultaneously through marker-assisted selection. Three major QTL hotspots, controlling multiple traits were identified on chromosome 1 (52.23-61.18 Mb), 2 (2.52-11.43 Mb) and 3 (1.32-3.95 Mb). The identified genomic regions and underlying candidate genes can be utilized in pyramiding favorable alleles for improving source-sink relationships in sorghum under diverse environments.

Analysis of corn and sorghum flour mixtures using laser-induced breakdown spectroscopy.

BACKGROUND: In a world constantly challenged by climate change, corn and sorghum are two important grains because of their high productivity and adaptability, and their multifunctional use for different purposes such as human food, animal feed, and feedstock for many industrial products and biofuels. Corn and sorghum can be utilized interchangeably in certain applications; one grain may be preferred over the other for several reasons. The determination of the composition corn and sorghum flour mixtures may be necessary for economic, regulatory, environmental, functional, or nutritional reasons. RESULTS: Laser-induced breakdown spectroscopy (LIBS) in combination with chemometrics, was used for the classification of flour samples based on the LIBS spectra of flour types and mixtures using partial least squares discriminant analysis (PLS-DA) and the determination of the sorghum ratio in sorghum / corn flour mixture based on their elemental composition using partial least squares (PLS) regression. Laser-induced breakdown spectroscopy with PLS-DA successfully identified the samples as either pure corn, pure sorghum, or corn-sorghum mixtures. Moreover, the addition of various levels of sorghum flour to mixtures of corn-sorghum flour were used for PLS analysis. The coefficient of determination values of calibration and validation PLS models are 0.979 and 0.965, respectively. The limit of detection of the PLS models is 4.36%. CONCLUSION: This study offers a rapid method for the determination of the sorghum level in corn-sorghum flour mixtures and the classification of flour samples with high accuracy, a short analysis time, and no requirement for time-consuming sample preparation procedures. © 2020 Society of Chemical Industry.

Genetic architecture of kernel composition in global sorghum germplasm.

BACKGROUND: Sorghum [Sorghum bicolor (L.) Moench] is an important cereal crop for dryland areas in the United States and for small-holder farmers in Africa. Natural variation of sorghum grain composition (protein, fat, and starch) between accessions can be used for crop improvement, but the genetic controls are still unresolved. The goals of this study were to quantify natural variation of sorghum grain composition and to identify single-nucleotide polymorphisms (SNPs) associated with variation in grain composition concentrations. RESULTS: In this study, we quantified protein, fat, and starch in a global sorghum diversity panel using near-infrared spectroscopy (NIRS). Protein content ranged from 8.1 to 18.8%, fat content ranged from 1.0 to 4.3%, and starch content ranged from 61.7 to 71.1%. Durra and bicolor-durra sorghum from Ethiopia and India had the highest protein and fat and the lowest starch content, while kafir sorghum from USA, India, and South Africa had the lowest protein and the highest starch content. Genome-wide association studies (GWAS) identified quantitative trait loci (QTL) for sorghum protein, fat, and starch. Previously published RNAseq data was used to identify candidate genes within a GWAS QTL region. A putative alpha-amylase 3 gene, which has previously been shown to be associated with grain composition traits, was identified as a strong candidate for protein and fat variation. CONCLUSIONS: We identified promising sources of genetic material for manipulation of grain composition traits, and several loci and candidate genes that may control sorghum grain composition. This survey of grain composition in sorghum germplasm and identification of protein, fat, and starch QTL contributes to our understanding of the genetic basis of natural variation in sorghum grain nutritional traits.

Changes in protein and starch digestibility in sorghum flour during heat-moisture treatments.

BACKGROUND: Heat-moisture treatment (HMT) has been used to modify properties of sorghum starches. However, information is limited on the effects of HMT on the digestibility of starch and the concurrent changes in protein in sorghum flour. The objectives of this research were to identify heat-moisture conditions to increase the resistant starch (RS) content of sorghum flour and investigate changes in sorghum proteins and starch structure. RESULTS: Sorghum flours with different moisture contents (0, 125, 200, and 300 g kg-1 w.b.) were heated at three temperatures (100, 120 and 140 °C) and times (1, 2 and 4 h). HMT of sorghum flour increased its RS level. The flour treated at 200 g kg-1 moisture and 100 °C for 4 h had a high RS content (221 g kg-1 vs. 56 g kg-1 for the untreated flour). Starch was not gelatinized when sorghum flours heated at moisture content of 200 g kg-1 or below. Sorghum protein digestibility and solubility decreased during HMT. The increase in RS of sorghum flour upon HMT was attributed to enhanced amylose-lipid complexes and heat induced structural changes in its protein fraction. CONCLUSION: HMT can be used to increase RS content in sorghum flour without gelatinizing its starch, thereby providing sorghum flour with unique food applications. © 2017 Society of Chemical Industry.

Presence of tannins in sorghum grains is conditioned by different natural alleles of Tannin1.

Sorghum, an ancient old-world cereal grass, is the dietary staple of over 500 million people in more than 30 countries in the tropics and semitropics. Its C4 photosynthesis, drought resistance, wide adaptation, and high nutritional value hold the promise to alleviate hunger in Africa. Not present in other major cereals, such as rice, wheat, and maize, condensed tannins (proanthocyanidins) in the pigmented testa of some sorghum cultivars have been implicated in reducing protein digestibility but recently have been shown to promote human health because of their high antioxidant capacity and ability to fight obesity through reduced digestion. Combining quantitative trait locus mapping, meta-quantitative trait locus fine-mapping, and association mapping, we showed that the nucleotide polymorphisms in the Tan1 gene, coding a WD40 protein, control the tannin biosynthesis in sorghum. A 1-bp G deletion in the coding region, causing a frame shift and a premature stop codon, led to a nonfunctional allele, tan1-a. Likewise, a different 10-bp insertion resulted in a second nonfunctional allele, tan1-b. Transforming the sorghum Tan1 ORF into a nontannin Arabidopsis mutant restored the tannin phenotype. In addition, reduction in nucleotide diversity from wild sorghum accessions to landraces and cultivars was found at the region that codes the highly conserved WD40 repeat domains and the C-terminal region of the protein. Genetic research in crops, coupled with nutritional and medical research, could open the possibility of producing different levels and combinations of phenolic compounds to promote human health.

High-throughput micro-plate HCI-vanillin assay for screening tannin content in sorghum grain.

BACKGROUND: The HCI-vanillin assay is a well-accepted method for determining tannin content in sorghum but is limited to small sample sets due to the time-consuming nature of the method. The objective was to develop an accurate and repeatable high-throughput 96-well plate assay for breeders to screen large sample sets of sorghum for tannin content. Validation of the high-throughput assay was tested on 25 sorghums suspected to contain tannin. RESULTS: Approximately 30 measurements per day were completed using the conventional assay compared to 224 measurements using the 96-well platform. The correlation between the two tannin assays was 0.98. The coefficient of variation (CV) was 3.54% and 3.21% for the 96-well and conventional method, respectively. The 96-well assay exhibited good repeatability, with the inter-plate CV between 2.77% and 4.85%. CONCLUSION: The high-throughput 96-well HCI-vanillin assay exhibited an eightfold increase in the number of measurements completed and was as accurate as the conventional HCI-vanillin assay.

Transcriptome and metabolome analyses reveal regulatory networks associated with nutrition synthesis in sorghum seeds.

Cereal seeds are vital for food, feed, and agricultural sustainability because they store and provide essential nutrients to human and animal food and feed systems. Unraveling molecular processes in seed development is crucial for enhancing cereal grain yield and quality. We analyze spatiotemporal transcriptome and metabolome profiles during sorghum seed development in the inbred line 'BTx623'. Morphological and molecular analyses identify the key stages of seed maturation, specifying starch biosynthesis onset at 5 days post-anthesis (dpa) and protein at 10 dpa. Transcriptome profiling from 1 to 25 dpa reveal dynamic gene expression pathways, shifting from cellular growth and embryo development (1-5 dpa) to cell division, fatty acid biosynthesis (5-25 dpa), and seed storage compounds synthesis in the endosperm (5-25 dpa). Network analysis identifies 361 and 207 hub genes linked to starch and protein synthesis in the endosperm, respectively, which will help breeders enhance sorghum grain quality. The availability of this data in the sorghum reference genome line establishes a baseline for future studies as new pangenomes emerge, which will consider copy number and presence-absence variation in functional food traits.

Grain yellowness is an effective predictor of carotenoid content in global sorghum populations.

Identification of high carotenoid germplasm is crucial to assist breeders in provitamin-A biofortification of sorghum (Sorghum bicolor [L.] Moench). High-performance liquid chromatography is the gold standard for carotenoid quantification, however, it is not feasible for large scale phenotyping due to its high cost and low throughput. In this study, we tested the feasibility of using grain color as a high-throughput method of carotenoid biofortification breeding. We hypothesized that visual, color-based selection can be an effective strategy to identify high-carotenoid accessions. Yellow grain had significantly higher carotenoid content than red, brown, and white grain. The degree of yellowness could distinguish the presence or absence of carotenoids, but could not distinguish carotenoid concentrations within yellow-only accessions. The degree of luminosity of the grain, however, was able to better predict carotenoid concentrations within yellow-only accessions. Genome-wide association studies identified significant marker-trait associations for qualitative and quantitative grain color traits and carotenoid concentrations near carotenoid pathway genes-ZEP, PDS, CYP97A, NCED, CCD, and LycE-three of which were common between grain color and carotenoid traits. These findings suggest that using grain color as a method for screening germplasm may be an effective high-throughput selection tool for prebreeding and early-stage breeding in carotenoid biofortification.

Impact of heat and high-moisture pH treatments on starch digestibility, phenolic composition, and cell bioactivity in sorghum (Sorghum bicolor L. Moench) flour.

Sorghum (Sorghum bicolor L. Moench), characterized by substantial genetic diversity, encompasses some lines rich in health-promoting polyphenols. Laboratory studies have demonstrated anticancer properties of sorghum phenolics; however, their presence may impact nutritional factors, such as digestible starch. The objective of this study was to determine the effects of pH and high-moisture heating on starch digestibility, phenolic profile, and anticancer activity in sorghum. High Phenolic sorghum flour line SC84 was combined with buffer solutions (pH 3, 4, 5, 7, and 8) and heated for 0, 10, 30, 60, or 120 min. Starch digestibility was assessed using the K-DSTRS kit from Megazyme. Changes in phenolic composition were analyzed using total phenolic content (TPC) and condensed tannin content (CTC) assays coupled with reversed phase high performance liquid chromatography (RP-HPLC) analysis. Anticancer potential against human colorectal cancer cells (HCT116 and SW480) was determined though cell viability assay. Results indicated a significant increase in total starch digestibility of sample after heating. Heating samples for 10 min did not significantly reduce TPC of samples. However, CTC was significantly reduced with heating time, while pH exhibited no significant effect on CTC. The measured 3-deoxyanthocyanidins experienced a significant decrease (p < 0.0001), while certain flavonoids increased significantly (p < 0.05) after heating for 30 min or longer. Notably, the 10 min heating duration minimally affected anticancer activity, whereas longer heat times diminished extract efficacy against human colorectal cancer cells. Alkaline pH levels significantly decreased anticancer activity, regardless of heating time. Importantly, heating sorghum for 10 min improved starch digestibility with minimal compromise to potential health benefits. These findings suggest promising implications for the development of high-phenolic sorghum products, and provide valuable insights to guide forthcoming animal and clinical studies. The demonstrated impact of wet-heating on increased starch digestibility, coupled with the preservation of phenolic content and bioactivity, underscores the potential of incorporating high-phenolic sorghum lines in future functional food formulations.

Variability in tannin content, chemistry and activity in a diverse group of tannin containing sorghum cultivars.

BACKGROUND: Tannins are large polyphenolic polymers and are known to bind proteins, limiting their digestibility, but are also excellent antioxidants. Numerous studies investigating the functional properties of sorghum tannin have been conducted by comparing grain samples from different sorghum lines without considering the other intrinsic characteristics of the grain. The purpose of this study was to remove the confounding intrinsic factors present in the endosperm so the effect of the tannins could be evaluated utilizing a unique decortication/reconstitution procedure. RESULTS: The tannin content of the 14 cultivars tested ranged from 2.3 to 67.2 catechin equivalents. The bran fractions were studied for their impact on protein binding and antioxidant capacity. Protein digestibility by pepsin ranged from 8% to 58% at the highest tannin level addition. Protein binding ranged from 3.11 to 16.33 g blue bovine serum albumin kg⁻¹ bran. Antioxidant capacity ranged from 81.33 to 1122.54 µmol Trolox equivalents g⁻¹ bran. High-performance size-exclusion chromatography detailed molecular size distributions of the tannin polymers and relationship to tannin functionality. CONCLUSION: The tannin content and composition play a significant role in determining tannin functionality. These differences will allow for selections of high-tannin sorghums with consideration of the biological activities of the tannins.

Optimization of tannin-containing sorghum bran addition to gluten-free bread.

Ingredients used to enhance sensory quality of gluten-free (GF) bread often lack in nutrients. This presents nutritional challenges for celiac-positive individuals and fails to meet expectations of healthfulness for non-celiac GF consumers. Sorghum (Sorghum bicolor L. Moench) flour can provide acceptable GF bread properties, and tannin-containing varieties contain antioxidants concentrated in the bran along with dietary fiber. Using a central composite design, tannin-containing sumac sorghum bran, gum (xanthan + guar), and water levels were optimized in a GF sorghum-based bread formulation. Loaf specific volume and gas cells/cm2 were maximized while minimizing hardness and cell wall thickness. The optimum formulation containing 14.2% sorghum bran, 1% gum, and 145% water (flour basis) effectively increased dietary fiber in bread to 13.4% (considered "high fiber") and showed oxygen radical absorbance capacity of 61.6 µmol TE/g. This optimum formulation did not differ from a sorghum flour-based control bread in consumers' (N = 100) liking of color, texture, flavor, overall acceptability, nor willingness to buy (WTB). All mean hedonic scores (numbered 9-point scale) were above 5, whereas average WTB was 4.7 for the optimum formulation and 4.6 for the control (9-point Likert scale) among consumers varying in GF bread consumption habits. Perceived bread bitterness was low (averaging 2.85 on 9-point intensity scale), did not vary between samples despite marked differences in antioxidant capacity, and was not correlated with WTB. When utilizing effective optimization models with key functional ingredients, sumac sorghum bran addition can enhance dietary fiber and antioxidant potential in sorghum-based GF breads without compromising quality attributes.

Effect of Flour Particle Size on the Glycemic Index of Muffins Made from Whole Sorghum, Whole Corn, Brown Rice, Whole Wheat, or Refined Wheat Flours.

The unique properties of sorghum are increasingly being studied for potential health benefits, with one area of emphasis being the impact of sorghum consumption on mitigating type 2 diabetes. The glycemic index (GI) of muffins made from whole grain sorghum flour ground to three different particle sizes (fine, intermediate, coarse) was tested on eight healthy volunteers (ages 18-40) and compared to the glycemic index of whole grain corn, wheat, and rice flours produced using a similar product formula. Sorghum flour ground through a 0.5 mm screen ("fine") had an overall similar particle size to that of the brown rice flour ground using a 0.5 mm screen. The range of GI values was 32 to 56, with only the GI of intermediate milled sorghum flour being lower than that of corn, rice, or wheat (p < 0.05). The lowest glycemic index (32 +/- 17) was found when using sorghum flour with an intermediate particle size (167 +/- 4 µm). Muffins made using brown rice had the next lowest glycemic index at 37 +/- 17. All GI values calculated had large standard deviations, which is common for these types of studies. These results can assist in the product development process to advance the quality of healthy, gluten-free sorghum-based foods for consumers. Further research should investigate if these results can be duplicated and the possible reason for the lower GI of intermediate particle size sorghum flour.

Variations in Total Protein and Amino Acids in the Sequenced Sorghum Mutant Library.

Sorghum (Sorghum bicolor) is the fifth most important cereal crop worldwide; however, its utilization in food products can be limited due to reduced nutritional quality related to amino acid composition and protein digestibility in cooked products. Low essential amino acid levels and digestibility are influenced by the composition of the sorghum seed storage proteins, kafirins. In this study, we report a core collection of 206 sorghum mutant lines with altered seed storage proteins. Wet lab chemistry analysis was conducted to evaluate the total protein content and 23 amino acids, including 19 protein-bound and 4 non-protein amino acids. We identified mutant lines with diverse compositions of essential and non-essential amino acids. The highest total protein content in these lines was almost double that of the wild-type (BTx623). The mutants identified in this study can be used as a genetic resource to improve the sorghum grain quality and determine the molecular mechanisms underlying the biosynthesis of storage protein and starch in sorghum seeds.

Performance of a Handheld MicroNIR Instrument for Determining Protein Levels in Sorghum Grain Samples.

Near infrared (NIR) spectroscopy is widely used for evaluating quality traits of cereal grains. For evaluating protein content of intact sorghum grains, parallel NIR calibrations were developed using an established benchtop instrumentation (Perten DA-7250) as a baseline to test the efficacy of an adaptive handheld instrument (VIAVI MicroNIR OnSite-W). Spectra were collected from 59 grain samples using both instruments at the same time. Cross-validated calibration models were validated with 33 test samples. The selected calibration model for DA-7250 with a coefficient of determination (R2) = 0.98 and a root mean square error of cross validation (RMSECV) = 0.41% predicted the protein content of a test set with R2 = 0.94, root mean square error of prediction (RMSEP) = 0.52% with a ratio of performance to deviation (RPD) of 4.13. The selected model for the MicroNIR with R2 = 0.95 and RMSECV = 0.62% predicted the protein content of the test set with R2 = 0.87, RMSEP = 0.76% with an RPD of 2.74. In comparison, the performance of the DA-7250 was better than the MicroNIR, however, the performance of the MicroNIR was also acceptable for screening intact sorghum grain protein levels. Therefore, the MicroNIR instrument may be used as a potential tool for screening sorghum samples where benchtop instruments are not appropriate such as for screening samples in the field or as a less expensive option compared with benchtop instruments.

Modulation of kernel storage proteins in grain sorghum (Sorghum bicolor (L.) Moench).

Sorghum prolamins, termed kafirins, are categorized into subgroups α, ß, and γ. The kafirins are co-translationally translocated to the endoplasmic reticulum (ER) where they are assembled into discrete protein bodies that tend to be poorly digestible with low functionality in food and feed applications. As a means to address the issues surrounding functionality and digestibility in sorghum, we employed a biotechnology approach that is designed to alter protein body structure, with the concomitant synthesis of a co-protein in the endosperm fraction of the grain. Wherein perturbation of protein body architecture may provide a route to impact digestibility by reducing disulphide bonds about the periphery of the body, while synthesis of a co-protein, with known functionality attributes, theoretically could impact structure of the protein body through direct association and/or augment end-use applications of sorghum flour by stabilizing ß-sheet formation of the kafirins in sorghum dough preparations. This in turn may improve viscoelasticity of sorghum dough. To this end, we report here on the molecular and phenotypic characterizations of transgenic sorghum events that are down-regulated in γ- and the 29-kDa α-kafirins and the expression of a wheat Dy10/Dx 5 hybrid high-molecular weight glutenin protein. The results demonstrate that down-regulation of γ-kafirin alone does not alter protein body formation or impacts protein digestibility of cooked flour samples. However, reduction in accumulation of a predicted 29-kDa α-kafirin alters the morphology of protein body and enhances protein digestibility in both raw and cooked samples.

Effects of protein digestion on in vitro digestibility of starch in sorghum differing in endosperm hardness and flour particle size.

In vitro digestibility of starch in sorghum grains differing in endosperm hardness and flour particle size was investigated. The starch digestibility increased as the particle size of flour decreased, but no clear trend was observed in digestibility of starch in sorghum flours milled from grains with different hardness. The protein matrix affected the digestion of starch. The pH value (2.0 vs. 1.3) was a critical factor affecting protein digestion. Optimum pH (pH 2.0 for pepsin) digested more protein, resulting in a greater digestion of starch. Resistant starch (RS) content was 8.5-26.3% in isolated sorghum starch but higher (10.6-29.5%) in sorghum flours. Protein digestibility decreased after cooking while starch digestibility increased compared to native sorghum flours; disulfide bonds formed between protein molecules. RS content of cooked sorghum flour was much higher without pepsin treatment (16.93-23.99%) than that of cooked sorghum flour with pepsin treatment (4.86-12.53%).