However, excessive sulfur dioxide will pollute the environment and cause deterioration of food quality, and even the high residual levels can increase the risk of cancer. As Gastrodia elata Blume is prone to corruption during processing, sulfur fumigation is often used for preservation. In this study, spectral analysis and Texture Profile Analysis (TPA) were used to investigate the effects of traditional sulfur fumigation processing on the morphology quality, edible quality and structural characteristics of G. elata. The results showed that compared with direct drying, the pH decreased by 0.399 of the sulfur fumigated after steamed treatment G. elata, and the morphology quality, pasting ability and gel edible quality of the starch were significantly improved. In addition, it was suggested that sulfur fumigation after steaming could promote the release of molecular chains from starch granules and thus enhance the cross-linking between molecules, which explained the reason for the improve of starch edible quality. This study can provide technical and theoretical support for improving the quality of starch rich foods, replacing sulfur fumigation and reducing potential environmental hazards.The sulfur dioxide gas (SO It has the function of relieving wind and spasm, calming liver Yang, dispelling wind and dredging collaterals. It is traditionally used as treatment for manage convulsions in children, epilepsy, tetanus, headache and dizziness, hand and foot failure, numbness of limbs, rheumatism, and arthralgia  was collected in December 2021 from Xiaocaoba Town, Zhaotong, Yunnan, China , for use in this study. Sulfur was purchased from Sinopharm Chemical Reagent Co., Ltd., . The steaming equipment is an electric cooker . All chemical reagents used in this study were of analytical grade.Fresh G. elata was cleaned, it was divided into steam treatment (St) and nonsteam treatment (NSt). In steam treatment, G. elata is steamed in a steamer for 30 min. After drying the surface moisture, G. elata was removed for sulfur fumigation treatment (Sf) and nonsulfur fumigation treatment (NSf) from the St and NSt treatments. The Sf treatment according to the ratio of G. elata:sulfur (20:1) and placed in a fumigation box (60 cm × 45 cm × 35 cm). After the sulfur was ignited, the fumigation box was capped, and a small hole of approximately 1 cm2 was left for ventilation. After fumigation for 90 min, it was removed. All G. elata were placed into the electric constant temperature blast drying oven  with 50°C hot air and dried for 5 days, and the final water content was approximately 5%. After drying, the G. elata was powdered and passed through the 200 mesh sieve. The starch was extracted following the method of Jiang et al. . The sulfur dioxide residue was determined following the method of Kang et al. . Solubility and swelling power were determined following the method of Lu et al.  with a sM1 is the starch mass in the supernatant after drying, M2 is the sample mass, M3 is the sediment mass after centrifugation, S is the sample solubility.where G. elata starch, mix it with 5 mL of 0.5 mol/L KOH solution, dilute it 10-fold, take 1 mL of solution, adjust the pH to 3.5 with 0.1 mol/L HCl, add 0.1 mL iodine reagent , and measure the absorbance at 625 nm. Calculate the content according to the following standard curve:Amylose content following the method of Uarrota et al.  with a sY is the amylose content (%) in the mixed standard solution, and X is the absorbance measured at 625 nm.Use the mixed solution with amylose content  to measure the absorbance at 625 nm to obtain the standard curve. Where G. elata starch was stirred in a boiling water bath for 1 h to prepare a 2% starch suspension. After cooling at 25°C, the transmittance was measured at 620 nm.Paste transparency was determined following the method of Correia et al. . G. elatG. elata starch and water to prepare a 10% aqueous starch solution, shake and mix in a boiling water bath for 30 min. Then, the sample was refrigerated for 12 h at 4°C. After the formation of the gel, it was removed and restored to room temperature. Texture analyzer  was used for measurement, and P/0.5 was used as the probe. The parameters were set as follows: the test speed was 1 mm/s, the test distance was 13 mm, and the trigger force was 2 g. Hardness, adhesiveness, springiness, cohesiveness, gumminess, chewiness, and resilience were recorded.Texture characteristics were determined following the method of Lu et al.  with a sG. elata starch and dried KBr and scan them in the wavenumber range of 500–4,000 cm–1 at room temperature after tablet pressing.Following the method of Lu et al. , the FT-Following the method of Lu et al.  with a sG. elata starch by rapid viscosity analyzer . Dissolve 1.25 g of G. elata starch in 25 mL of water, stir for 30 min, mix well, and measure it according to the method of rice configuration file in RVA.Following the method of Li et al.  with a sG. elata starch were determined by STA . The test temperature was 30°C–600°C, nitrogen was the ambient gas, and the rising temperature was 10°C/min.Following the method of Ma et al. , the the–1. Data were fitted by the power law equation:The rheological properties of the samples were determined using a rheometer . The test was conducted at 25°C using 40 mm parallel plates. Take 0.25 g of starch to prepare a 5% starch suspension (mix it evenly in boiling water for 10 min and cool it to room temperature), then transfer it to a rheological plate and balance it for 3 min. According to the method of Lu et al. , frequen–1), γ is the shear rate (s–1), and n is the non-Newtonian exponent.where σ is the shear stress (Pa), K is the consistency coefficient  to prepare 5 mg/mL in 0.2 M NaOH solution, hydrolyzed at 120°C for 1 h, centrifuged at 12,000 rpm for 10 min, and a 0.22 μm microporous filter membrane was used, and then the sample was transferred into a 1.8 mL injection vial. 0.05 M NaCl solution was used as the mobile phase for analysis at a flow rate of 0.6 mL/min. BRT105-104-102 series gel column (8 × 300 mm), column temperature: 40°C, injection volume: 20 μL, detector: differential detector RI-10A. A standard curve was drawn with the retention time (RT) of dextran standards on the lgMw-RT of the correction curve equation:lgMn-RT of the correction curve equation:Y is lgMw, and X is the retention time (RT).In (1), Y is lgMn, and X is the retention time (RT).In (2), Following the method of Zeng et al. . Add 7.520 is the glucose content in the solution at 20 min of enzymatic hydrolysis, G120 is the glucose content in the solution at 120 min of enzymatic hydrolysis, FG is the free sugar content in the starch solution at 0 min, TS is the total mass of the starch sample used, Y is the glucose content (mg), and X is the absorbance measured at 540 nm.GThe microstructure of the starch powder was observed with SEM , and the sample powder was fixed on the sample rack and placed in the instrument for observation.AFM  was used to observe the microscopic morphology of the starch paste. One hundred milligrams of the sample was added to 20 mL of distilled water, stirred and mixed in a boiling water bath, and the paste was continued for 10 min and diluted with hot water to 50 μg/mL. The sample was placed on the mica surface and observed after drying.Following the method of Ma et al. with a slight modification . SamplesP < 0.05) under Tukey’s test. All values are expressed as the mean ± standard deviation. Drawing was performed with Prism 9.0 .All experiments were repeated at least in triplicate. Data were analyzed using SPSS 26.0  for statistical significance  was 223.67 and 356.67 mg/kg, respectively, which indicate that the samples had been successfully pretreated by sulfur fumigation. The sulfur dioxide residue of G. elata with St Sf treatment was 1.59-fold that with NSt-Sf treatment, which indicates that the steamed G. elata more easily absorbed sulfur dioxide than the nonsteamed G. elata .After St treatment, the amylose content of G. elata . The pasG. elata, the whiteness of starch was significantly increased compared with that of NSt treatment , 2,933 cm–1 (C-H stretching), 1,643 cm–1 (C-O bending), 1,384 cm–1 (C-H bending), and 1,024 cm–1 (C-O stretching). A weak absorption peak was generated between 1,176 and 1,024 cm–1, which was caused by the tensile behavior of C-C and C-O. The weak absorption peak near 1,383 cm–1 was attributed to the bending vibration of -C-H, C-C-H, and C-O-H. Compared with CK , G. elata starch did not produce new absorption peaks after St treatment or Sf treatment, indicating that steaming or sulfur fumigation treatment did not alter the functional groups of G. elata starch  of G. elata starch decreased, while PV and FV increased. Compared with G. elata starch treated with NSt-NSf and NSt-Sf, the Pt of the starch treated with St-NSf and St-Sf decreased by 11.7 and 15.47°C, respectively. The Sf treatment also reduced Pt and promoted the increase in PV and FV. Compared with starch treated with NSt-NSf and NSt-Sf, the Pt of starch treated with St-NSf and St-Sf decreased by 0.48 and 4.24°C, respectively, and the PV and FV of St-Sf increased by 1.95 and 1.33-fold, respectively, compared with St-NSf  and final viscosity (FV), but neither NSt-NSf nor NSt-Sf-treated starches reached PV and FV. Therefore, the PV and FV of St h St-NSf .Gastrodia elata starch exhibits two rapid weight loss stages at 50–100°C and 250–350°C. The weight loss in the first stage is caused by the evaporation of crystal water in starch, while the second stage is attributed to the decomposition of starch. The presence of only two weight loss stages indicates that starch is a single component except water  were significantly higher than those of the nonfumigation treatment (NSt-NSf and St-NSf), indicating that the degree of molecular crosslinking of gelatinized G. elata starch solution was also significantly enhanced following sulfur fumigation.The storage modulus (G′) and loss modulus (G′′) gradually increase with increasing angular frequency, and G′ > G′′, indicating that weak gel . The G′ G. elata starch eluted at 22–34 min, and the low-molecular-weight amylose eluted at 34–45 min  was 1.27 and 1.49-fold higher; the weight-average molecular mass (Mw) was 1.30 and 1.53-fold higher; and the polydispersity index (PDI) was 1.03 and 1.02-fold higher, respectively. After Sf treatment, the peak time of G. elata amylopectin elution was earlier than that of NSf-treated amylopectin. Compared with NSt-NSf and St-NSf, NSt-Sf and St-Sf increased by 0.34 and 0.33 min; Mn decreased by 17.55 and 2.94%; Mw decreased by 18.34 and 4.14%; and PDI decreased by 1.09 and 1.60%, respectively  was finer and irregular  induced by sulfur fumigation is significantly higher than the pH increase (0.578) due to steaming, resulting in a relatively small effect of steaming on pH.Nonetheless, we also observed a very interesting phenomenon. Without sulfur fumigation, the pH of e et al.  and Jorge et al.  reportedG. elata starch promoted by steaming and sulfur fumigation. However, steaming and sulfur fumigation decreased the granule size of gelatinized starch. Compared with sulfur fumigation, steaming has a more significant effect on increasing the transparency of starch paste. However, sulfur fumigation can further consolidate and enhance the effect of steaming on the transparency of starch paste. It can be seen that the combination of steaming and sulfur fumigation can enhance the whiteness and paste transparency of G. elata starch.Paste transparency is affected by the color of starch and the size of granules in the solution. The lighter the color and smaller the starch granules are, the higher the paste transparency . To incrG. elata starch were broken and polymerized after sulfur fumigation, resulting in an increase in final viscosity (FV) and chewability. This is consistent with the research conclusion of Xie et al. . Amylopectin α-1,6 glycosidic bonds form amylose after hydrolysis and debranching, and the unbranched chains become short amylose chains  and XRD can reflect the molecular structure and crystal structure of starch. In this study, there was no significant difference between the functional groups of standard starch (CK) and  methods , which i crystal . Moreove crystal . Sulfur e region . In addie region , type A G. elata starch by increasing the whiteness and paste transparency through the bleachability of sulfite but also cause the degradation of amylopectin to produce amylose and promote an increase in RSD content to increase its edible quality. In addition, the release degree of starch chains from starch granules was increased by sulfur fumigation after steaming, thus enhancing the pasting characteristics of gel and cooking qualities such as hardness, chewiness, and stickiness, which meant that the taste quality of starch was increased. This work explained the improvement mechanism of steaming and sulfur fumigation treatment on G. elata edible quality and provided a scientific basis for the rational use of sulfur fumigation technology in food processing. This modified starch method may be used to improve the gel taste of low viscosity food and the digestion rate of starch products.This study showed that sulfur fumigation after steaming could not only increase the morphological quality of The original contributions presented in this study are included in the article/JG, ZC, and YY wrote and reviewed the manuscript. TZ, JG, TX, and FW collected the samples. JG, ZC, LG, CW, and YY provided analysis ideas and reviewed the manuscript. JG and ZC reviewed the manuscript and drew the picture. LG and XC supervised the work and read the manuscript. All authors contributed to the article and approved the submitted version.


The present study investigated the effect of incorporating 10, 20, and 30% apple powder obtained by freeze-drying, and apple powder produced with osmotic pre-treatment in sugar beet molasses solution, into muffins. The powder was freeze-dried and introduced as a whole spelt wheat flour replacement in muffins. The obtained products were investigated for their chemical composition and technological properties, and were subjected to a sensory analysis as well as a consumer acceptance test. Increasing the substitution level from 0 to 30% apple powder lowered the protein, starch, and fat content, while moisture content, sugar, and cellulose showed the opposite trend. The sensory analysis results indicated that the addition of apple powder or apple powder with osmotic pre-treatment (apple OT+Lyo powder) to the ingredients of muffins positively affected the taste, smell, mastication, and appearance of the final product. Consumers rated the muffins with 30% apple OT+Lyo powder as the most acceptable. Principal component analysis, an artificial neural network, and global sensitivity analysis were utilized to differentiate among muffin samples, and to estimate the corresponding influence of the substitution of spelt flour with apple powder or apple OT powder on the observed quality and nutritional parameters of the muffins. Muffins are favored for breakfast and snack food due to their ready-to-eat form, low price, tender texture, and sugary taste ,2. The pFibre  can be added into baked products as a replacement for traditional ingredients such as flour, sugar, or fat ,14. In aThe application of osmotic treatment (OT) as a pre-treatment for fruit powder production enhances the powder’s quality, nutrient composition, and energy efficiency . FurtherSpelt wheat is an old subspecies of traditional wheat with technological properties that are slightly diverse from common wheat, and is usually viewed as a flawed grain . NeverthUsing principal component analysis (PCA) enables differentiation among the investigated muffin samples and a graphical presentation of relations between observed parameters. The effects of substituting flour with apple powder and OT apple powder on the chemical composition, technological characteristics, and color of muffins was examined using global sensitivity analyses based on the developed ANN model. Artificial neural networks (ANN) in diverse implementations usefully contribute to the engineering, technological processes, and development of food products .The present study focused on the exploitation of apple powder prepared by freeze-drying (Lyo apple powder) and apple powder obtained with osmotic pre-treatment in sugar beet molasses and freeze-drying (OT+Lyo apple powder), in varying levels , as a functional ingredient in whole spelt wheat muffins. In addition, the effects of both apple powder varieties’ incorporation on muffins’ chemical composition and technological and sensorial properties were studied. A consumer acceptance test was performed in order to select the muffins with the best properties.in.s, SG 2.624g/gin.s.) were stored in a freezer at −18 °C for 24h. The frozen apples were freeze-dried using a freeze dryer . The samples were placed on metal trays, and the freeze-drying parameters were set to a pressure of 1.6 Pa and condenser temperature of −57 °C. The FD process was performed as described by Nowak and Jakubczyk, 2020 [Granny Smith apples were acquired at a nearby market in Novi Sad (Serbia) and kept at 4 °C before use. This apple variety was selected due to its acidic, sharp, and juicy tart flavor, which provides enhanced sensory attributes to the muffin samples. The initial moisture content was 85.48%. The osmotic treatment was performed in sugar beet molasses (pH 8.2 and 80% dry matter), as described by Nićetin et al., 2017 . Water lyk, 2020 .w/w). Dried samples were finely ground into a powder, using a universal laboratory mill type WZ-1 .The Lyo apple powder was obtained by freeze-drying, while OT+Lyo apple powder was obtained via the hybrid drying method. For preparation of OT+Lyo apple powder, the apple samples were dried applying a freeze drier (FD) and coupling this action with osmotic treatment of sugar beet molasses prior to the FD step; this was performed until the final moisture content was 12.03%  of muffins with Lyo apple and OT+Lyo apple powder were analysed.Each muffin sample was measured for its specific weight and specific volume 24 h after baking, according to the AACC method . A ScannThe texture of the muffins was determined instrumentally on a texture analyzer TA.XTplus . The hardness and springiness of the muffins were measured using a 36-millimetre cylinder probe (P/36R) and a 5-kilogram load cell. Instrument settings were as follows: pre-test speed, test speed, and post-test speed were 2.0 mm/s, 1.0 mm/s, and 10.0 mm/s, respectively. The muffins were sliced off the top crust in order to obtain the soft inside of the muffin, and eachmuffin was perforated for testing. The muffins were evaluated at 25% compression strain. Hardness (g) was defined as the force required to compress the product by a present distance of 25%. A simple way of looking at the springiness property is to record the force after 30 s, divide this by the maximum force and multiply by 100%. The tests were performed using six replicates per batch.The muffin color was measured using a tri-stimulus color meter type CR-400  equipped with D65 illuminant. The results were expressed as per CIELab system in terms of coordinates: L*—lightness , a*—redness , and b*—yellowness ; W—whiteness is an attribute by which an object is judged to approach the preferred white. The measurements were observed under constant lighting conditions, at 28 °C, using the color attributes of a white control plate, L* = 98.76, a* = −0.04, and b* = 2.01 .ΔL*—difference in L* parameter between control and muffin sample with apple powder addition;Δa*—difference in a* parameter between control and muffin sample with apple powder addition;Δb*—difference in b* parameter between control and muffin sample with apple powder addition.Colour variations between the control muffin sample and muffin samples with apple powder and OT apple powder (ΔE), were determined by the following equation:Sensory evaluation of muffins with apple and osmotically treated apple powder was conducted as described by Filipović et al. 2016  and ISO With the intention of testing the consumer acceptability of muffins with apple and OT apple powder, a total of 250 untrained assessors were engaged in this examination. All respondents have consented to participation in the study. Consumers performed the test in order to assess the quality and acceptability of muffins. Sensory parameters  of the muffins with the addition of apple powder and osmotically treated apple powder were rated on a 9-level hedonic scale  .p ≤ 0.05 significance levels. PCA analyses, an ANN model, global sensitivity analysis, and correspondence analysis of the acquired results were conducted by applying StatSoft Statistica 10.0® software.Descriptive statistical analyses using one-way ANOVA of the results (displayed as the mean ± standard deviation—SD) were conducted with StatSoft Statistica 10.0 software. The significant differences were resolved according to the post hoc Tukey’s HSD (honestly significant differences) test at The color plot diagram was designed with R software v.4.0.3 (64-bit version) with the “circle” method, upper type.w—weight coefficient in ANN models, i—input variable, j—output variable, k—hidden neuron, n—number of hidden neurons, m—number of inputs.Yoon’s global sensitivity formula for the obtained ANN model was exploited in order to evaluate the relative influence of the input parameters on output variables, based on weight coefficients of the established ANN models :(2)RIg compared with the control sample (1). Conversely, an increase in the quantity of Lyo+OT apple powder added to muffins  leads to a statistically significant decrease in hardness (compared between samples 5–7).The hardness of a muffin sample is expressed as the maximum peak force at the first bite, and depends mainly on moisture and fat content . By incrOsmotic dehydration cause changes to the cell walls of the apple itself, which is reflected in significantly lower hardness of the muffin (samples 5–7) compared to the control sample (1) and compared to the samples with the apple powder (2–4). The springiness of the muffins refers to the rate of deformation recovery at which a deformed muffin sample shifts back to its undeformed shape after the involved deforming tension is withdrawn . With a As one of the crucial factors for consumers’ acceptance of a product, color is a primary concern in valuing consumers’ opinions, quality features, and acceptability of a food product . The addp ≤ 0.01; r = 0.919, p ≤ 0.01 and r = 0.966, p ≤ 0.01, respectively), and negatively correlated to the contents of cellulose and sugars . Furthermore, the flour content is positively correlated to the specific volume , and negatively correlated to moisture content and colour coordinate a* . The apple content is correlated negatively to flour content.p ≤ 0.05; r = 0.888, p ≤ 0.01, respectively), and also negatively correlated to the content of moisture, sugars, and cellulose .The protein content is positively correlated to starch and fat content, and to specific volume .The starch content is negatively correlated to fat content  and positively correlated to fat content .The fat content is negatively correlated to moisture .The cellulose content is negatively correlated to starch and fat content  and negatively correlated to the content of starch and fat .The sugar content is positively correlated to moisture content , while the content of Na is positively correlated to the content of Ca and K .The content of K is positively correlated to the content of Mg and Ca .The specific volume is negatively correlated to cellulose content .The hardness of muffin samples was negatively correlated to the contents of Ca, K, and Na  and positively correlated to the hardness of muffin samples .The colour coordinate L* of muffin samples was negatively correlated to the contents of Mg, Ca, K, and Na  and positively correlated to moisture and sugar content .The colour coordinate a* of muffin samples was negatively correlated to the contents of starch and fat  and positively correlated to the colour coordinate L* of muffin samples .The colour coordinate b* of muffin samples was negatively correlated to the contents of Mg and K  and positively correlated to the colour coordinates L* and b* of muffin samples .The colour coordinate W of muffin samples was negatively correlated to the contents of Mg, Ca, K, and Na  added to the muffin dough positively affected the majority of the chemical composition responses while negatively affecting technological characteristic responses. The incorporation of nutritively rich and also cellulosic material consequently provided a negative effect on the finished product’s total quality. Similar results were reported by Filipovic et al. , where tIn order to adequately describe the configuration of the results that would provide insight to the appearance of the muffin samples, PCA was exploited, and the results are exposed in The separation among control and samples containing Lyo apple and OT+Lyo apple powder could be discovered from the PCA biplot, The first two PCs outlined 80.76% of the total variance in the experimental data. The factors’ projection suggested that specific volume, L*, b*, and W mainly contributed to the first principal component, PC1 . The specific weight, hardness, and springiness contributed more to the second principal component, PC2 . The differentiation between samples could be recognized from the PCA graphic, where the samples with the higher amount of apple powder are arranged on the left side of the graphic. In comparison, the samples with a lower amount of apple powder are arranged on the right side of the graphic.2 , and depleted values of the sum of squares (SOS). The applied training algorithm was BFGS 274, with a hyperbolic tangent for hidden activation and identity for output activation function. The ANN model was complex, with 179 weight biases for observed muffin samples. In order to comprehensively describe the impact of input variables on the ascertained output variables, the addition of flour and apple powder and treatment of the muffins’ chemical composition and technological parameters were analysed using Yoon’s global sensitivity equation in line with the weight coefficients of the developed ANN model [The substitution of flour with the Lyo apple powder and OT+Lyo apple powder on the chemical composition, technological characteristics, and color of the muffins was examined using an ANN model. The acquired optimal neural network model expressed an acceptable generalization capability for the experimental data, and could correctly anticipate the output parameters of the muffin samples for a range of the observed input parameters. In conformity with the ANN performance, the optimal number of neurons in the hidden layer for the chemical composition, technological characteristics, and color parameters calculation was 7 (network MLP 5-7-18), in order to attain elevated values of coefficient of determination, rNN model ,52.Following the Yoon presentation method of a declared ANN model, the graphical appearance of the ANN model results is displayed in According to In In 2 was 145.26; df = 84; p < 0.00004). The first two dimensions explained 90.16% of the total inertia, utilizing a substantially satisfactory quota of the raw information.The graphical appearance of correspondence analysis for experimental results exhibited in From According to The results revealed that spelt muffins with the addition of osmotically pre-treated apple powder were nutritionally enriched compared to the control muffins. Furthermore, by comparing the chemical composition of muffin samples, it was observed that the addition of osmotically pre-treated apple powder increases moisture, cellulose, and sugar content, as well as Ca, K, and Na content as a consequence of the mass transfer from sugar beet molasses. These results open an innovative aspect in which an agro-industrial side product, such as sugar beet molasses, could be utilized as a promising osmotic solution with the possibility of raising the nutritional properties of baked products.The percentage addition of apple powder (10 to 30%) and osmotically treated apple powder affects the reduction in the specific volume of the finished product without affecting specific weight. Moreover, it alters the muffin’s textural attributes by decreasing the muffin samples’ hardness and springiness.Yoon’s interpretation method performed on a created ANN model recognized the addition of osmotically pre-treated apple powder and osmotic treatment as the most influential parameters that affected the observed parameters.Based on sensory evaluation of the muffins, percentage increases in apple powder and osmotically treated apple powder positively affect taste and smell, while slightly reducing the appearance grade of the product.According to the consumer test results, the sample with the highest content of osmotically dehydrated apple was the one that achieved the highest scores. It can be concluded that the addition of apple powder osmotically pre-treated in sugar beet molasses to muffin-like bakery products as a flour supplement could be recommended as a valuable ingredient due to its nutritional and sensory properties.


Eight NQT phase I metabolites were found that had been formed by N-demethylation, oxidation, hydroxylation, and reduction. In addition, three reactive electrophiles, two aldehydes, and one iminium ion were identified, and the corresponding bioactivation mechanisms were proposed. The reported side effects of NQT may be related to the generation of reactive metabolites. Based on a literature review, this may be the first study of in vitro phase I metabolites, detailed structural characterizations, and NQT reactive intermediates.Tyrosine kinase inhibitors (TKIs) are very efficient for the treatment of EGFR-mutated lung cancer and show improved therapeutic efficacy. However, treatment with both first- and second-generation TKIs results in acquired resistance and is related to various toxicities; the EGFR T790M mutation has been associated with this resistance. Naquotinib  is a novel third-generation epidermal growth factor receptor tyrosine kinase inhibitor that has been shown to be more potent than osimertinib in the management of L858R plus T790M mutations. However, its bioactivation may occur and promote the formation of reactive electrophiles that are toxic. We hypothesize that these reactive intermediates are potentially involved in the side effects of NQT. Reactive metabolites are often formed by phase I metabolic reactions and cannot be characterized directly as they are transient in nature. Using liquid chromatography-tandem mass spectrometry (LC-MS/MS), we screened for  in vitro metabolites of NQT formed during incubation with human liver microsomes (HLMs) and then evaluated the generation of reactive electrophiles using capturing agents.LC-MS/MS was used to screen for  Epidermal growth factor receptor (EGFR) signaling pathway was recently identified as an important potential NSCLC therapeutic target.6 Tyrosine kinase inhibitors (TKIs), which regulate the EGFR, are very efficient for the treatment of EGFR-mutated lung cancer and show improved therapeutic efficacy. First-line TKIs regulating EGFR  show good initial responses against tumors harboring these EGFR mutations.7,8 However, treatment with these inhibitors results in acquired resistance in 60% of patients and is related to various toxicities,9,10 thereby reducing their therapeutic efficacies.11,12 Accordingly, second-generation irreversible EGFR TKIs  have also been developed.13,14 Unfortunately, NSCLCs have been shown to exhibit resistance to both first- and second-generation EGFR-TKIs within the first year of treatment.15 Notably, the EGFR T790M mutation has been identified in approximately 50% of NSCLC cases showing resistance to first- and second-generation EGFR-TKIs.8,13Non-small cell lung cancer (NSCLC) is a common lung cancer subgroup,13,14 Naquotinib  is a novel third-generation EGFR-TKI that was found to be more potent than osimertinib against T790M plus L858R mutations (T790M + L858R). Moreover, NQT and osimertinib have been shown to have a wide therapeutic window and comparable efficacy for cells with EGFR exon 20 insertions.16Third-generation TKIs have maintained the benefits of second-generation drugs by blocking mutant EGFR and overcoming resistance induced by the T790M mutation.17–19 Reactive electrophiles are electron deficient and can modify DNA and proteins by establishing covalent bonds that are considered the first step in drug-mediated organ toxicities.20,21 Monitoring of reactive metabolite production is a critical task in studying drug-induced toxicity. Reactive metabolites are often formed by phase I metabolic reactions and cannot be characterized directly as they are transient in nature. Instead, a capturing nucleophile can be used for reactive intermediates to trap the adducts formed, which are stable and can be characterized by liquid chromatography-tandem mass spectrometry (LC-MS/MS).22,23Metabolic detoxification involves pathways that transform xenobiotics and endogenous compounds into more hydrophilic molecular species to facilitate excretion outside the human body. The generated metabolites are usually less toxic than the parent molecules. However, in rare cases, bioactivation may occur and promote the formation of reactive electrophiles that are more toxic.24–27 The acryloylpyrrolidine moiety can undergo oxidative dealkylation, forming aldehyde intermediates that can be captured using methoxyamine.28 Glutathione and its derivatives are highly nucleophilic and react poorly with strong electrophiles. However, the iminium ion and aldehydes are electrophiles that can be trapped using potassium cyanide and methoxyamine, respectively.17,24,25 The adducts formed by nucleophilic–electrophilic interactions are considered stable and can be separated and identified by LC-MS/MS.22–24,29,30 We hypothesize that these reactive intermediates are potentially involved in the reported side effects of NQT.The International Union of Pure and Applied Chemistry name of NQT is 5-[(1-acryloyl-3-pyrrolidinyl)oxy]-6-ethyl-3-({4-[4-(4-methyl-1-piperazinyl)-1-piperidinyl]phenyl}amino)-2-pyrazinecarboxamide . Its chein vitro metabolites of NQT formed during incubation with human liver microsomes (HLMs) and then evaluated the generation of reactive electrophiles using capturing agents.Accordingly, in this study, we used LC-MS/MS to screen for 2.2.1.2O) was obtained from the Milli-Q plus filtration system .All chemicals and solvents were of analytical grade. NQT was purchased from Med Chem Express . Acetonitrile (ACN), pooled human liver microsomes , potassium cyanide and formic acid were procured from Sigma-Aldrich . HPLC-grade water  with electrospray ionization (ESI) source. Separation of the components of the metabolic mixtures was performed on a C18 column . The column temperature was adjusted at 23 ± 2 °C, and we used a gradient mobile phase at a flow rate of 0.2 mL min−1 and consisting of 10 mM ammonium formate in H2O  and ACN (solvent B). The stepwise-gradient system involved solvent B , solvent B , solvent B , and solvent B , with a post time of 15 min. The sample injection volume was 5 μL and had a total run time of 75 min, with the mass parameters optimized for vandetanib. Fragmentation of NQT, phase I metabolites, and related adducts were performed in the collision cell by collision-induced dissociation (CID). Detection was performed on a mass detector operated using a positive ESI source.14,15 Drying gas (N2) was used as at a flow rate of 12 L min−1, and collision gas (high-purity N2) was used at a pressure of 60 psi. Source temperature, capillary voltage, fragmentor voltage, and collision energy were set to 350 °C, 4000 V, 135 V, and 22 eV, respectively. Mass Hunter software (Agilent Technologies) was used to for instrument management and data acquisition.Separation and characterization of 2.3.in vitro metabolites of NQT was performed by incubating 30 μM NQT with 1.0 mg mL−1 HLMs in phosphate buffer  and 3.3 mM MgCl2. The incubation time and temperature were 2 h and 37 °C, respectively. The metabolic incubation reactions were performed in a shaking water bath. Initiation of the NQT metabolic reaction was performed by adding NADPH (1.0 mM) and quenched by the addition of 2 mL ice-cold acetonitrile, which was also used as a protein precipitating agent. Precipitates were removed by centrifugation at 9000 × g , and the supernatants were transferred to clean tubes and evaporated to dryness. Residues were reconstituted in mobile phase solvent. Ten microliters of each reconstituted sample was injected and analyzed using LC-MS/MS.35,36The screen for 2.4.m/z peaks were utilized to locate expected in vitro metabolites in the total ion chromatograms. Fragmentation suspected peaks using product ion (PI) mode were used to confirm the metabolite chemical structure by reconstructing daughter ions (DIs). The fragmentation pattern (FP) of NQT was used as a lead for interpretation and confirmation of the proposed chemical structures of in vitro metabolites and reactive intermediates generated in the metabolism of NQT.After full MS scanning over a selected range, extracted ion chromatograms of targeted The same HLM incubation with NQT was performed in the presence of the capturing agent (methoxyamine or potassium cyanide) to trap bioactive intermediates. To confirm the outcomes, all experiments were repeated three times, with appropriate controls.3.3.1.m/z 563 generated many DIs. Three DIs at m/z 463, 323, and 70 represented qualitative soft points inside the NQT structure (The NQT molecular ion peak (MIP) was found at 37.8 min in the daughter ion chromatogram DIC; . Fragmentructure .3.2.N-demethylation, oxidation, hydroxylation, and reduction. One cyano and two methoxyamine adducts were identified . Oxidative dealkylation of the acryloylpyrrolidine group formed an aldehyde that was captured by methoxyamine, forming NQT566. Both oximes were stable and identified using LC-MS/MS . Aldehyd4.N-Demethylation, oxidation, hydroxylation, and reduction metabolic reaction generated eight phase I NQT metabolites. Oxidative dealkylation of the acryloylpyrrolidine group formed aldehydes that were captured by MeONH2, forming NQT566 and NQT580. Hydroxylation of the piperidine ring in NQT, followed by dehydration created an iminium electrophile that was captured by KCN, forming NQT588. Bioactivation resulted in the formation of one cyano adduct and two methoxyamine adducts of NQT (s of NQT . These fThe authors declare no conflicts of interest.


Massive irreparable rotator cuff tears in patients for whom arthroplasty is not an option can be a challenging clinical scenario for shoulder surgeons to manage. To achieve the best patient outcomes, a myriad of options has been presented in the literature, including debridement with biceps tenotomy or tenodesis, various tendon transfer procedures, superior capsular reconstruction, biceps tendon rerouting, bursal acromion resurfacing, balloon spacers, and tuberoplasty. While debridement with biceps tenotomy and superior capsular reconstruction have historically provided improvements in patient-reported outcomes, high rates of arthritis progression and failure of graft healing have been noted with these techniques, respectively. The superior capsular reconstruction has also proven to be technically challenging. The biologic tuberoplasty procedure was developed after several studies noted a lack of correlation between graft healing and improvement in patient-reported outcomes in superior capsular reconstructions, as long as the tuberosity remained covered with the graft. We present a technically efficient and expedited technique using an acellular human dermal allograft. Arthroscopic biologic tuberoplasty for irreparable rotator cuff tears: an expedited technique. Pain and dysfunction are thought to derive from pathoanatomic uncoupling of normal glenohumeral joint mechanics. This is generally observed with large rotator cuff tears, leading to proximal migration of humeral head and its bony articulation with the acromion during arm elevation and rotation.Biologic tuberoplasty is indicated in nonpseudoparalytic patients with preserved glenohumeral articular cartilage with massive irreparable rotator cuff tears. These patients have good passive range of motion with intact or repairable subscapularis and are typically poor candidates for arthroplasty due to young age or medical comorbidities. X-ray findings typical of an irreparable rotator cuff tear include superior migration of the humeral head or an acromiohumeral (AH) distance <7 mm.The patient is placed in a lateral position using a bean bag with all bony prominences well padded. The arm is placed in a sterile arm positioner, which allows the surgeon to manipulate the arm intraoperatively. Longitudinal traction is applied to the arm to allow for opening of the subacromial space and glenohumeral joint. The arm is sterilely prepped and draped in the usual fashion.A diagnostic arthroscopy is performed using standard posterior and anterior portals. The lateral portal is established, and the subacromial space is then entered. A subacromial decompression is performed as needed, with care taken to preserve the coracoacromial (CA) ligament. The rotator cuff is examined arthroscopically and manipulated to ensure that it is irreparable and that partial repair may be performed. Concomitant procedures, such as biceps tenodesis and distal clavicle excision, are also performed as deemed appropriate.Following this, the area to be covered by the graft is measured in anterior-posterior and medial-lateral directions using an arthroscopic ruler. We prefer for the area of coverage to extend from the articular margin laterally past the edge of the greater tuberosity. The 3-mm thick acellular dermal allograft ArthroFLEX  is then prepped by an assistant on the back table. The ArthroFLEX has two physically distinct sides: reticular and papillary; the rougher reticular side with its larger pores is to be the inferior surgical side. Note, the ArthroFLEX is packaged with papillary side visible through clear side of packaging. To ensure maintenance of orientation of the graft, mark the papillary (smoother and superior) side with sterile marker immediately after opening inner pouch. In preparation of the graft, we arthroscopically measure anterior to posterior (AP) defect and undersize the graft by 3-5 mm (15-20%) allowing for slight stretching of the graft A. Note, Following this, an arthroscopic burr is used to prepare a bleeding bone bed with gentle decortication on the greater tuberosity. Anterior and posterior 2.6 self-punching knotless FiberTak anchors  are percutaneously, through separate portals, placed medially just off the articular margin . The repThe patient is placed in an abduction sling for 6 weeks postoperatively. Physical therapy starts at 7-10 days postoperatively and is initially geared toward pain reduction and passive range of motion with pendulum, Codman’s exercises, and periscapular function and activation. We allow weight bearing on the operative arm, as tolerated, and then allow active-assisted/active range of motion, as tolerated with strengthening to begin at 6 weeks postoperatively with a goal of pain-free range of motion by 3 months.,The concept of a biologic tuberoplasty stemmed from previous research on the superior capsular reconstruction (SCR), which has proven to be technically difficult to reproduce. Several recent articles have demonstrated radiographic graft healing in 38-81% of cases, with the majority of failures occurring on the glenoid side.,Tendon transfers have also been proposed as options for irreparable rotator cuff tears, based on tear location and functional deficits.Another recent technique involves insertion of a saline-filled balloon into the subacromial space to act as a bumper and a humeral head depressor. Despite several small case series reporting benefit, Metcalf et al. recently published a double-blinded randomized controlled trial of 117 patients, which demonstrated greater improvements in the Oxford Shoulder Score in a debridement/biceps tenodesis group only vs. those who received an identical procedure with insertion of the subacromial balloon.The above technique, as described, aims to provide pain relief through arthroscopic placement of a biologic bumper between the greater tuberosity and the acromion, which can be performed expeditiously and efficiently in patients who are poor candidates for arthroplasty or more extensive arthroscopic procedures. In such cases, the bumper will help to maintain a degree of humeral head depression and aid in deltoid retraining by relieving pain. This technique is reproducible and efficient, and we believe it will result in similar rates of graft healing and pain control compared to Mirzayan’s technique.Rehabilitation may be further expedited with this procedure, as compared to repair of a massive tear, tendon transfer, or superior capsular reconstruction. Although our preliminary results have been excellent, there is currently no long-term data on the procedure, and surgeons should proceed with this in mind.


Vaccination clearly decreases coronavirus disease 2019 (COVID-19) mortality; however, they also impose selection pressure on the virus, which promotes the evolution of immune escape variants. For example, despite the high vaccination level in especially Western countries, the Omicron variant caused millions of breakthrough infections, suggesting that the highly mutated spike protein in the Omicron variant can escape antibody immunity much more efficiently than the other variants of concern (VOCs). In this study, we investigated the resistance/susceptibility of T helper cell responses that are necessary for generating efficient long-lasting antibody immunity, in several VOCs. By predicting T helper cell epitopes on the spike protein for most common HLA-DRB1 alleles worldwide, we found that although most of high frequency HLA-DRB1 alleles have several potential T helper cell epitopes, few alleles like HLA-DRB1 13:01 and 11:01 are not predicted to have any significant T helper cell responses after vaccination. Using these predictions, a population based on realistic human leukocyte antigen-II (HLA-II) frequencies were simulated to visualize the T helper cell immunity on the population level. While a small fraction of this population had alarmingly little predicted CD4 T cell epitopes, the majority had several epitopes that should be enough to generate efficient B cell responses. Moreover, we show that VOC spike mutations hardly affect T helper epitopes and mainly occur in other residues of the spike protein. These results suggest that lack of long-lasting antibody responses is not likely due to loss of T helper cell epitopes in new VOCs.The online version contains supplementary material available at 10.1007/s00251-023-01295-8. The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has caused over 6 million deaths worldwide  T cell epitopes in silico and investigate the effect of vaccine selection pressure on epitope conservation and mutations in VOCs. Simulating a population with realistic HLA frequencies, we visualize the T helper cell immunity on the population level with respect to SARS-CoV-2 (and other human coronaviruses). With these results, we hope to gain insight into the potential need for a new (booster) vaccine to parallel SARS-CoV-2 immune evasion in the future.In this study, we aim to study spike  and binding affinity (BA). The EL-trained model includes the likelihood of a peptide being produced in the cell and naturally presented by an HLA-DRB1 molecule, whereas the BA-trained model focuses on peptide-HLA binding affinity only. As expected, the EL-trained model predicted less coronavirus spike epitopes than the BA-trained model  , the total number of predicted spike epitopes was corrected for protein length. The number of T helper cell epitopes per 100 amino acids for 14 most common HLA-DRB1 molecules ranged between 10 and 15 for the coronaviruses Fig. B. SARS-C+ T cell epitopes. Without correcting for this, H3N2 and H1N1 hemagglutinin had 5 and 6 predicted epitopes for HLA-DRB1 13:01, which is higher than the number of epitopes for SARS-CoV-2 WT . Omicron BA.2 retained more conserved epitopes than BA.1, likely because of its different mutation profile. Overall, there was no excessive loss of original T helper cell epitopes in the VOCs, as reported by Tarke et al .+ T cell epitopes per individual seem to decrease with the recent VOCs, which might point to a possible escape from the immunity built up in the last 2.5 years. However, the median number of T helper cell epitopes remain high.Because it is unclear whether subtle T helper cell epitope losses Fig. B–C have + T cell epitopes when infected with Omicron BA.1 . Therefore, we have used the African American HLA-DRB1 allele frequencies  vaccine is based on , we found that most mutations occurred in spike regions that are predicted as non-epitopes , indicating that predicted T helper cell epitopes are mainly present in conserved parts of the spike protein. In conclusion, SARS-CoV-2 spike mutations clearly occur less frequently in T helper cell epitopes. There might be two reasons explaining this observation. First, high polymorphism of HLA class II molecules in the population blocks the adaptation of the virus to escape T helper cell epitopes. Second, MHC molecules in general bind preferably hydrophobic peptides that are less likely to mutate  was calculated. Interestingly, mutations occurred more often in non-epitopes than in T helper cell epitopes . Luckily many of the alleles we studied are also rather common in other ethnicities. For example, the alleles we used in this analysis makes up 57% in African and Hispanic populations and 44% in Asian populations . Moreover, predicted continuous B cell epitopes covered a much smaller portion of the spike protein than the T helper cell epitopes. All these factors are, currently, limiting to study the evolution of B cell epitopes computationally. Still, we observed that even in continuous B cell epitopes, there are significantly more mutations than in CD4 T cell epitopes (results not shown), suggesting that there might be a strong selection pressure on SARS-CoV-2 to escape human B cell responses.Unfortunately, it is still a big challenge to be able to predict B cell epitopes. When we used the best available tool to predict continuous epitopes, DiscoTope,  SARS, Middle East respiratory syndrome (MERS), and 229E and NL63 coronavirus spike protein sequences were retrieved from UniProt   Below is the link to the electronic supplementary material.


There is substantial literature interrogating the application of donor derived Tregs for the prevention of graft versus host disease (GVHD). This Mini-Review will focus on the recipient’s Tregs which persist post-transplant. Although treatment in patients with low dose IL-2 months post-HSCT are encouraging, manipulating Tregs in recipients early post-transplant is challenging, in part likely an indirect consequence of damage to the microenvironment required to support Treg expansion of which little is understood. This review will discuss the potential for manipulating recipient Tregs in vivo prior to and after HSCT . Strategies that would circumvent donor/recipient peripheral blood harvest, cell culture and ex-vivo Treg expansion will be considered for the translational application of Tregs to improve HSCT outcomes.Human and mouse CD4 Their use in experimental as well as clinical transplants has shown the ability to promote engraftment and diminish both host versus graft (HVG) and graft versus host (GVH) responses  was not a focus of the study , 12. Folt origin Table 1 t origin , 17. Immt origin . In the in vivo. Early studies by Shlomchik and colleagues found that radiation-resistant recipient T cells ameliorated GVHD in a chronic murine model  post-HSCT which possess functional suppressive capacity ne model . Experim T cells . Studies T cells . The res disease , 19. Othpartment , 21. Qu partment . In studpartment . Studiesin situ evidenced their activated and functional state and hypothesized that these host T cells contributed to tissue GVHD  and chronic GVHD patients, although the origin of these Tregs were not examined . As described above, studies from a number of laboratories have shown that rTregs persist following a variety of conditioning levels including some ablative strategies and transplant and hence could be manipulated as well , 18, 20.in vivo, reviewed in  into patients’ months post-HSCT with diagnosed cGVHD, increased their peripheral blood Treg levels and function which correlated with reduced cutaneous GVHD  in the lamina propria of the large intestine 4 days post-allo-HSCT which persisted up to 10 days  and level of conditioning (myeloablative vs reduced intensity) will impact the effectiveness of pre- and post-transplant Treg stimulation. For example, following aggressive conditioning and allo-HSCT, Treg manipulation may be delayed due to an impaired microenvironment needing time to reconstitute. In contrast, targeting of the rTreg compartment prior to transplant can circumvent these events enabling potent expansion of these cells. Furthermore, the presence of increased levels of activated effector T cells post-transplant likely would decrease the selective targeting of Tregs by the reagents discussed. Lastly, in contrast to applying the beneficial effects of only peripheral blood dTregs, treatment of recipients is not restricted to circulating populations but includes all tissues comprising key targets of GVHD.via augmenting DNP-kinase activity, promoting DNA repair pathways  are likely to differentially disrupt the microenvironment as well as influence the kinetics of its repair and rebound. Based on our own work, we postulate that 3-4 weeks is minimally needed to “rejuvenate/re-build” the Treg microenvironment under moderately aggressive conditioning regimens. Although identifying the key signals can ultimately lead to efforts to minimize/protect the microenvironment from conditioning induced damage, developing strategies to target Tregs prior to conditioning currently provides an excellent opportunity to exploit the rTregs before environmental disruption and early findings suggest such manipulations can improve overall survival and function early post-HSCT.All Tregs present in recipients post-transplant are subject to the extant microenvironment which nurtures and maintains homeostasis of the compartment. Our laboratory and others have struggled to successfully manipulate Tregs SC and RL outlined and wrote the review article. DW, BP, HB, and VP read, edited and critiqued the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by funds from the National Institutes of Health , and the Sylvester Comprehensive Cancer Center (RBL). SC is the recipient of a ASTCT New Investigator Award. Research reported in this publication was also supported by the National Cancer Institute (Award P30CA240139).The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.RL is a compensated consultant/advisory board member for and equity holder in Heat Biologics and consultant for Kimera Labs. Neither are directly relevant to this review.The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Small and medium-sized enterprises (SMEs) are pushed to introduce new technologies due to different requirements and changes in the business setting. The SMEs' transformation to exploit new technologies is challenging given their lack of resources and the complexity of technological transformation, which encompasses technology assimilation and business model innovation (BMI). Although studies recognize the complementarity of technology assimilation and BMI for benefiting from technology, the literature is fragmented, and the technological transformation phenomenon remains abstract, especially in the SMEs' context. To improve understanding of technological transformation in SMEs, a systematic literature review was performed on 165 peer-reviewed papers published from 1999 to 2022, building upon BMI and technology assimilation constructs. The descriptive analysis outlines the field's evolution in terms of research and technological trends. The content analysis shows that: most papers focus on factors; the literature falls short of providing theoretical conceptualization and guidelines for the technological transformation process; only a few studies are dedicated to assessing the outcomes of technological transformation in SMEs; and the papers suggest that SMEs present a low transformation level. Finally, we inductively built a framework for technological transformation and suggest five research avenues. The current business setting characterized by environmental changes, new requirements, technological development, and new opportunities pushes SMEs to rethink their traditional production methods and introduce new technologies to remain competitive over time , with recent literature reviews contributing to overall technological transformation literature , including 165 peer-reviewed papers. More specifically, the SLR research aims to provide the state-of-the-art of the research field of technological transformation in SMEs, building upon technology assimilation and BMI insights. The SLR draws on descriptive and content analysis. The descriptive analysis presents the development of the study field (from 1999 to 2022). The content analysis inductively assesses the current stage of literature on technological transformation by explicitly merging BMI and technology assimilation concepts. Moreover, we classify the papers' foci into research streams and identify drivers, barriers, enablers, control variables, processes, and outcomes for technological transformation. Finally, we derive a framework for technological transformation based on content analysis.This work contributes to advancing the understanding of the technological transformation phenomenon in SMEs by shedding light on the existing contributions and future research avenues in the field. As technological transformation is a research endeavour dispersed across different research fields, we contribute to the literature as a first step to make it explicit as a concept and to derive insights into the interactive merger between technology assimilation and BMI. This paper is structured as follows. Section This study adopted a systematic literature review (SLR) as a research approach to ensure a transparent, structured, and replicable process, reducing bias and enhancing the data analysis's legitimacy . We identified two literature reviews  were selected because they are the most comprehensive multidisciplinary bibliographic databases  SMEs as technology providers and not as users; (ii) BMI not related to technology; (iii) mention technology-related terms or technology commercialization without addressing technology introduction/use;We want to provide an overview of technological transformation by connecting technology assimilation and BMI; thus, we discarded papers that do not address both concepts.The articles' selection considered the following filters: (i) elimination of duplicates; (ii) reading of title, abstract, and keywords; and (iii) reading of the full text. The use of EndNote® software facilitated the organization of the retrieved papers. The following criteria were used for screening the articles:The search for papers was performed in November 2020, and the databases recovered 352 papers (209 from Scopus and 143 from WOS), of which 269 were not duplicates. Using the screening criteria, we read the papers' title, abstract, and keywords (resulting in 126 articles), followed by reading the full text (resulting in 90 articles). After that, an update was performed (in December of 2022), using the exact keywords and screening criteria limited to 2020 to 2022 years to update until 2022 (resulting in 75 new papers). The selection resulted in 165 articles , and the remainder is theoretical. Most empirical studies are qualitative (56%), followed by quantitative studies (41%) and mixed methods (3%). Section Regarding the technology type, most of the papers refer to digital technologies (31%), e-commerce (14%), and industry 4.0 (12%). The publications suggest technological trends over time; for instance, there is a concentration of recent papers on digitalization (45% of the articles published from 2019 to 2022) and industry 4.0 (14% of the papers published from 2018 to 2022). These technological trends align with the fourth industrial revolution that demands SMEs to adopt these technologies. Moreover, the role of digital technologies was emphasized during the COVID-19 crisis since they enabled SMEs to keep their operation running with the new restrictions  (around 80%). Until 2014, the number of publications was not higher than three publications per year. Some possible explanations for the recentness of the papers are: BMI is a relatively recent literature outgrowth  and Technological Forecasting and Social Change (five publications). The prevalence of papers in the Sustainability journal suggests the trend of studies discussing how technologies can generate sustainable opportunities and benefits for companies.Given the multidisciplinary of the field, we found journals with different scopes with three publications, namely: Journal of Open Innovation: Technology, Market, and Complexity, Industrial Marketing Management, Information Systems Frontiers, Journal of Business Research, Journal of Manufacturing Technology Management, and Journal of Small Business and Entrepreneurship.The most prolific authors are Julian Müller and Kai-Ingo Voigt (four publications), and their research concerns the 4.0 industry. Two other research clusters were identified: Harry Bouwman, Shahrokh Nikou, and Mark de Reuver, that have two publications in common, and their research is part of the European project “Envision”, which aims to support SMEs in BMI, and Anis Rozmi, Puteri Nohuddin, Abdul Hadi, Mohd Bakar that have two publications in common regarding ICT technologies. The papers were mainly conducted in Europe (52%), followed by Asia (33%), America (11%), Africa (3%), and Oceania (1%), especially in developed countries.Technology assimilation can be defined based on management or system perspectives  or as an outcome (innovative BM) in which changes are made in a BM's component  or the BM's architecture  are adequate for SMEs  value elements: value creation, value capture, and value offer; or (ii) building blocks: key resources, channels, customer segments, key partners, key activities, customer relations, revenue streams, cost structure, value proposition  technology providers or (ii) users. In the first case, technology providers promote changes in their BMs to offer technology feasibly by changing their value capture . For example, Olson et al.  is one driver often cited in the literature. Some relative advantages expected by SMEs are cost reduction, operational efficiency, and increased sales. Moreover, recent studies claim that technologies may positively impact sustainability; for instance, digital technologies may enable improvements in carbon emission, material consumption, and waste reduction . For example, Groot et al.  state thOrganizational drivers are mainly related to top management actions and vision. Firstly, top management can provide support and pursue strategic orientation toward technological transformation. SMEs can also be internally motivated to adopt technologies when the owner/manager perceives market opportunities or changes in the market (e.g. changes in consumer behavior) that new technology can address  or its process (i.e. transforming the business to exploit new technology). Indeed, the barriers vary according to the stage of technology assimilation  and are considered a barrier to adoption  Moeuf et al.  concludeSMEs generally do not adopt complex systems  lack of partners, (ii) behavior of partners, and (iii) relationship management. According to Kollmann and Häsel , alliancCertain technologies and BMI require collaboration to leverage benefits and fulfil functions; thus, partnership problems may be even more problematic for those cases. For example, Reim et al.  identifiThe lack of customer acceptance/demand is an external resistance to the technology use or related products and/or services. For example, Coreynen et al.  describeRegarding organizational barriers, the knowledge and perception of the owner-manager play a central role in decisions of technological transformation. The owner-manager can be unaware of new technologies available  the definition of innovation strategy; (ii) the commitment of resources for technological transformation; (iii) the culture towards technological transformation; and (iv) alignment of operational and strategic actions. The innovation strategy needs to encompass the conditions for using the technology and change the business  according to marketing opportunities. Top management should provide guidance, supervision, and training, especially in the first moment of technology introduction, to ensure that the employees can operate with new technology and do not commit repetitive mistakes  partnerships among companies to accomplish business functions within the value chain or (ii) external support. In the first case, connecting ecosystem partners entails risks such as interdependence and coordination challenges, necessitating BMs' alignment with the value network and strategic risk management , which depend on their context. In this regard, SMEs can achieve outcomes by combining different capabilities . In this relationship, BMI determines the nature of complementarity between BMs and paths for monetization. This complementarity includes value mechanisms such as value offer , value creation (identifying customers and how they are engaged) and value capture  , which is not commonly addressed in the literature. This relationship is focused on enabling SMEs to acquire/use technologies that were not previously feasible for these companies. Although this relationship is not often discussed in the literature, it is a topic that makes sense in the SMEs' context, given their lack of resources.The outcomes of technological transformation can be discussed in terms of (i) the level of technology assimilation, (ii) the level of BMI, and (iii) performance outcomes. The level of technology assimilation can be evaluated in terms of the stage of technology assimilation within a company. For example, Fichman and Kemerer  employ aThe level of BMI refers to the extent to which BM elements have been changed . Secondly, the context-dependence of technological transformation  hampers its general conceptualization, with a concentration of papers in some technological contexts. Thirdly, the research streams are also fragmented individually as technology assimilation and BMI can be discussed in terms of process or outcome, resulting in a complex combination of the fields due to the high level of abstraction.To improve the understanding of technological transformation in SMEs, we conduct an initial merge of technology assimilation and BMI fields in the context of introducing a technology by (i) classifying the papers' foci; (ii) identifying, unifying, and positioning the factors; (iii) identifying what is stated about the process, control variables and outcomes; and (iv) providing a conceptual framework.We identified six main papers' foci, the most prominent being factors. The concentration of papers in analyzing factors was expected since existing literature reviews on technology assimilation and BMI in SMEs also focus on factors  to meet SMEs' needs is in line with the integrative technological transformation perspective on introducing technology.Regarding the process of technological transformation, the analysis highlights the need for external support, especially in cases where technology is complex or new and the SME does not possess specific capabilities . Moreover, there is a lack of conceptualization of the technological transformation process that may be linked to its context dependence. This lack of conceptualization challenges SMEs since technological transformation combines two already challenging processes with interrelationships.The interrelationships between BMI and technology assimilation are not generally addressed in the literature, although the overlapping of the processes is suggested. From the analysis of the papers, we identified three relationships: (i) technology assimilation that drives BMI, (ii) BMI that mediates the relationships between technology assimilation and a company’s performance, and (iii) BMI as a precursor of technology assimilation. The last relationship is not usually addressed in the literature but emerged from the analysis since technology acquisition can be problematic for SMEs.The studies generally show a low level of transformation in terms of technology assimilation and/or BMI, and few studies evaluate the performance outcomes of technological transformation. This fact may be linked to the recentness of the papers and to SMEs' characteristics that lag in the technology adoption or experiment with a gradual transformation process compared to larger firms, which may result in the lack of studies on performance outcomes or the low level of outcomes reported. Finally, based on the content analysis, we derive the technological transformation framework Fig. .Fig. 2TeTo build this framework, we considered the overview of technological transformation: from its initiation to its outcomes. The drivers identified .Secondly, the phases of the technological transformation process remain abstract. The literature is not homogeneous in presenting phases for BMI  technological transformation paths for SMEs or (ii) BM patterns or configurations. For example, agri-food SMEs face pressure to comply with new trends in the food industry , which can be addressed by introducing food processing technologies Rodgers . In thisThe technology transformation studies were mainly conducted in developed countries, which are considered years ahead of their counterparts in terms of technological transformation  presenting the evolution of the field, the primary study approaches, and technological trends; (ii) classifying the paper's foci within the field; (iii) integrating and positioning technological transformation factors; (iv) inferring the interrelatedness between technology assimilation and BMI; and (iv) providing a framework of technological transformation.The findings indicate that technological transformation is a relatively new topic in the literature, and its combination remains abstract. The research domain is also concentrated in various technological contexts . In this sense, we develop a general framework for the subject, as the antecedents, processes, and outcomes were extracted from studies in various contexts.The findings demonstrate that most of the papers focus on factors. These factors were identified, integrated, and positioned in the technological transformation framework. The analysis of factors and processes of technological transformation suggests an overlap between technology assimilation and BMI, and we provide insight regarding their relationships (three relationships were identified). Results also show a limited number of studies assessing the outcomes of technological transformation in SMEs, and that, generally, SMEs present a low transformation level.Finally, the limitations in the keywords used to perform the paper's search impact this work. We used generic terms without considering keywords for specific technologies  since the scope of this paper is general. Secondly, we only selected papers that present the term “business model” since we were interested in papers that explicitly address the BMI concept. Thirdly, papers focusing on the individual streams (technology assimilation or BMI) were not selected since they are out of the scope. The authors’ subjectivity also limits the papers' analysis and classification because only the authors' viewpoints were contemplated.


Particle deposition on film cooling surface is an engineering issue that degrades the thermal protection of turbine blade. Here, we present a combined experimental and numerical investigation on the particle deposition in the vicinity of multiple film cooling holes to reveal the effect of interactions between cooling outflows on particle deposition. The numerical simulation of film cooling with a group of three rows of straight film cooling holes is conducted and validated by experimental data with blowing ratios ranging from 0 to 0.08. Wax particles with size range from 5 to 40 μm are added in the heated mainstream to simulate the particle deposition in the experiment. The simulation results show the decrease of particle deposition with blowing ratio and various deposition characteristics in different regions of the surface. The flow fields from numerical results are analyzed in detail to illustrate deposition mechanism of the particles in different regions under the interactions of cooling outflows. The cooling air from the holes in the first row reduces the particle concentration near the wall but causes particle deposition in or between the tail regions by the generated flow disturbance. The cooling air from the latter hole separates the diluted flow in the upstream from the wall, and creates a tail region without particle deposition. This revealed particle deposition characteristics under the effect of outflows interaction can benefit the understanding of particle deposition in engineering applications, where multi-row of cooling holes are utilized. For thermal protection of the turbine blade, cooling air from secondary air system is supplied through the elaborately arranged film cooling holes on the blade to form a thin cooling air film for reduction of heat transfer from the hot mainstream flow. This so-called film cooling technology has been widely used for decades in turbomachinery for its high reliability in turbine blade cooling. Numerous studies have been carried out to reveal the mechanism of film cooling, optimize the design of hole, and improve cooling efficiency ,3,4,5,6.One of the challenges facing in film cooling is the deposition of particles, which are mainly originated from the pollutants, e.g., sand, debris, and ash, in inlet air or fuel, on the turbine blade ,8,9. TheFor external particle deposition, Demling et al.  presenteMany studies also focused on the dynamics of the deposition process in film cooling by changing the flow parameters or hole geometry. Rozati et al.  simulateResearches on the particle deposition were mostly limited to the configuration of a single hole or a single row of holes, neglecting the interactions between the outflows from holes in different rows, which may play an important role in practical applications, as multiple rows of film cooling holes were utilized. This multiple row configuration aims not only for approaching the reality, but also for simulating the transpiration cooling, the extreme form of film cooling, by using the straight holes ,24. In tA low speed open loop wind tunnel is utilized for the experiment of the particle deposition. The schematic of the experimental setup is shown in F, which measures the mass flow ratio between the cooling air and hot air in the mainstream [F from 0 to 0.08 for both experiment and simulation. The cooling efficiency The blowing ratio instream , can be ciency η  is definx axis, a height of 15 mm along the z axis, and a length of 49 mm along y axis. Seven straight film cooling holes connect the coolant chamber and mainstream channel. The diameter of the cooling hole is 1 mm, and the distances between the holes in the flow direction y and the transverse direction x are both 3 mm, same as the experiment.In numerical simulation, we choose a representative unit, including seven straight film cooling holes, one in the center and the other six surrounding holes arranged in the corners of a hexagon. As illustrated in F of the case. Since this calculation intends to simulate the particle deposition in transpiration cooling, therefore the outflow angle between the film cooling hole and y-axis is set at 90F is varied between 0 and 0.08—the same as in transpiration cooling.The commercial software ANSYS ICEM is used to generate Hexahedral mesh in the fluid domain. O-grid mesh is applied in the region of film cooling holes for high mesh quality. The detail of the mesh is shown in 3) of the particles in this study; (3) the added particles do not affect the flow field; (4) the rebound of the particle caused by the surface roughness is neglected. Based on these assumptions, the particle is considered to be trapped on the wall when it contacts with the surface. If the particle is not colliding with the surface, its motion will be calculated until it leaves the controlled volume. Therefore, we are able to simulate the process of particle deposition in the mainstream channel. It has to be noted that the simulation is a steady state simulation by setting the mass flow rate of the added particle at To calculate the particle deposition and track the particle in the mainstream channel, discrete phase model (DPM) is implemented upon the flow field and heat transfer simulation. The trajectory of the particle is calculated using the Lagrangian method, which tracks the motion of each particle in the flow field. The motion of the particle is mainly determined by the flow field, but is also affected by its intrinsic random motion. To capture the motion of the particles and avoid the difficulties in coupled simulation caused by the interaction between the particles and flow field, several assumptions are made: (1) each particle is considered as a non-spinning hard sphere with uniform density; (2) the interaction between particles are neglected due to the low mass fraction  d–e. AlthIn the experiment, the particle deposition on the surface is a time-dependent process. The particles added in the mainstream would keep accumulating on the surface with time, resulting in an ever-increasing deposition thickness. However, we use steady state simulation for modeling the particle deposition. To validate our modeling result on particle deposition, we compare the particle capture efficiency F from both the experiment result and simulation data. The increase in F means more cooling air is supplied through the film cooling holes on the flat surface to prevent the particle deposition, leading to less deposition of the particles on the surface. The simulation result has a similar trend with the experimental data, although the simulation underestimates the particle deposition. The underestimation of The result in h in the flow direction. The irregular pattern of the deposited particles that appeared on the surface (y-axis with the center points lying on the lines s1 and s2 (y coordinate For the added particle, due to its higher density than that of surrounding gas, it intends to fall upon the surface under the gravitational force. Although the particle’s small size and high gas velocity in the mainstream induce the added particles to follow the gas and move through the channel, the low velocity field in the boundary layer near the wall makes the particle deposition possible. When the blowing ratio is zero, i.e., no cooling air is supplied through the film cooling holes in the wall, the particles collide with the surface and deposit on the wall under the gravity a. With t surface a may be 1 and s2 b, and plh of the deposited particle substantially decreases (F, the deposition thickness h generally decreases on the surface. For the convenience of the description, we denote the holes from “H1” to “H7”, and their tail regions, i.e., the regions behind the holes along the flow direction from “HT1” to “HT7”, as shown in h in HT1 and HT2 becomes larger than the surrounding region. However, the thickness h in the tail regions behind other film cooling holes, from H3 to H7, are significantly smaller than the surrounding regions, particularly when With the supply of the cooling air on the surface (ecreases a. The inTo obtain a quantitative result of the particle deposition on the surface when using h0 . The varh0 on s1 a, we obsh0 on s2 b, the vaTo obtain further insights into the particle deposition characteristics resulting from the interaction between outflows of multiple film cooling holes, we provide the detail of the flow fields from various view angles for further analysis.c in the In 1 and s2 e,f with  (F=0.02 a,c and F (F=0.08 b,d. HereIn “In ling air a,b. Alonx-axis before the film cooling holes (Based on these flow fields and concentration contours, we are able to understand the particle deposition characteristics under the interactions between the cooling outflows from the neighboring cooling holes. For the film cooling outflow, due to its low velocity, it enters into the boundary layer of the mainstream and induces the mixing in the mainstream, which not only dilutes the particle concentration in the boundary layer but also gives higher possibility of the particle deposition by the induced disturbance. Therefore, the film cooling outflow from one hole may not necessarily eliminate the particle deposition in its tail region a. The contration c,d, resuntration a.In conclusion, the particle deposition in the vicinity of multiple film cooling holes are presented by simulation and validated by experiment. In general, the supply of cooling air through the hole reduces the particle deposition, and this effect increases with the blowing ratio. The interactions between the outflows of the film cooling holes reveals different deposition characteristics in different regions of the surface. The particle deposition occurs after the hole in the first row, but it can be eliminated in the tail regions of the holes starting from the second row. The flow field analysis from simulation indicates that the interaction between the outflows of different holes generates such characteristics of particle deposition. Due to the high particle concentration in the boundary layer near the wall, the cooling air from the hole in the first row dilutes the concentration, but induces the disturbance that facilitates particle deposition. The cooling air from the hole in the second row lifts the diluted flow and separates the particles from the wall, generating a tail region without deposition. These revealed particle deposition characteristics could be helpful in understanding the deposition mechanism in practical applications and benefit the design of the film cooling and transpiration cooling.


Adlay starch has great potential as a cereal starch, but it has several weaknesses, namely a low swelling volume, low solubility, and low stability. The purpose of this study was to improve the characteristics of adlay starch, such as porosity, functional properties, and pasting properties, through starch modification using freeze moisture treatment (FMT) and ozonation. This study consisted of several treatments, namely FMT, ozonation, and a combination of FMT + ozonation. The results show that the FMT and ozonation generally increased water absorption capacity, swelling volume, solubility, and number of pores of the starch granule. The pasting properties showed an increase in the viscosity of the hot paste and caused a decrease in the gelatinization temperature, breakdown, and setback viscosity. FMT 70% + ozonation produced modified adlay starch with a porous granular surface, swelling volume value of 21.10 mL/g, water absorption capacity of 1.54 g/g, a solubility of 9.20%, and an increase in the amorphous structure but did not cause the emergence of new functional groups. The combination of FMT + ozonation was effective in improving the functional, pasting, and physicochemical properties of adlay starch. Adlay is one of the cereals that can be used as a source of starch that can be applied to various food and chemical industries ,2. StarcVarious methods for starch modification have been developed, namely chemical, physical, enzymatic, and microbiological modifications (fermentation) or a combination of several modification methods ,12,13. H3), known as ozonation. Ozone is a type of strong oxidizing agent but is safe for food products because it will not leave chemical residues as ozone will immediately turn back into oxygen after oxidizing the material [Modification of the FMT method can be combined with other methods to increase the effectiveness of improving the characteristics of adlay starch, such as oxidation with ozone , and then filtered. The dregs were immersed again in NaOH solution (0.3%) and then filtered up to two times. All the filtrate was deposited for one day, then washed with clean water, and then dried. The dried starch was then ground again and sifted at 100 mesh.Adlay starch was extracted by a combination of dry and wet methods . The driThe experiment was carried on with five modified adlay starches and a control (native adlay starch). Modified adlay starches included ozonated adlay starch, FMT adlay starch at 60% initial moisture content, FMT adlay starch at the initial moisture content of 70%, FMT adlay starch at the initial moisture content of 60% + ozonation, and FMT starch adlay at the initial moisture content of 70% + ozonation. FMT was carried out in thin-wall containers, frozen at −50 °C, and sublimated using a freeze dryer (Christ Alpha 1–4 LDplus) at −50 °C for 48 h. Meanwhile, the ozonation was carried out using an ozone generator  at a flow rate of 2 L/min for 20 min and stirred every 5 min .The surface morphology of starch was analyzed by SEM-JSM6510, JEOL Ltd. Tokyo, Japan. The sample was placed on the sample holder and then coated with gold. The morphology of the granules was read at magnification scales of 2000, 5000, and 7000.Swelling volume (SV) and solubility were analyzed by weighing out 0.7 g of starch, which was then put into a centrifuge tube, to which 25 mL of distilled water was added, and finally, stirring the solution using a vortex. The sample was heated using a water bath . The sample was immersed in cold water and then centrifuged to separate the precipitate (gel) from the supernatant. The gel and supernatant in the centrifugation tube were separated, and then the supernatant volume was measured. The supernatant was then dried in a constant cup using an oven, then weighed , and SV Water absorption capacity (WAC) was analyzed by weighing out one gram of adlay starch, to which 10 mL of distilled water was added, and then stirring the solution using a vortex, followed by incubated for 30–40 min and centrifugation to separate the precipitate from the supernatant. The supernatant was separated from the precipitate and weighed , and theA total of 3.5 g of starch was added to the distilled water (25 mL), and then a canister containing starch and aquadest was mounted on a Rapid Visco Analyzer (RVA-SM2). The test was carried out for 13 min, in which the temperature was raised to 95 °C and held for 7 min, and then the temperature was lowered to 50 °C and held for 5 min .Color analysis was carried out using the Color Flex EZ CFEZ 2508 chromameter instrument. The chromameter was first calibrated with the standard black and white plates contained in the tool. The starch sample was put in a glass cup and then placed in the sample holder, and the color chromaticity of the sample was read. Tests were carried out with the Hunter L*, a*, b*, and ΔE* color system.−1, a mid-infrared KBr beam splitter with a range of 11,000 to 375 cm−1, and XT KBr extended-range mid-infrared optics, combined with attenuated total reactance (ATR). The sample (1–2 mg) was added to pure KBr powder. The sample mixture was placed in the sample holder and then pressurized and maintained for several minutes. Then, the FTIR spectra were read, and the data were interpreted.Functional groups of starch were analyzed with a “Nicolet iS10 FTIR spectrometer” with a spectral range of 7800 to 350 cmp < 0.05; then, to determine the differences between treatments, this step was followed by Duncan’s multiple range test using PASW Statistics version 18.0 .The data obtained were analyzed for variance (ANOVA) with SV and solubility are affected by the noncovalent bonds of amylose and amylopectin. Starch granules will absorb water and swell to about 30% of the initial volume when starch is put into cold water. However, when starch granules are heated in excess water, it causes the breaking of hydrogen bonds and disruption of the crystalline structure so that starch granules expand. The SV of native adlay starch and adlay starch modified by FMT and ozonation is shown in p < 0.05). Meanwhile, the ozonation treatment without FMT did not increase SV significantly. The highest increase occurred in the starch modified by FMT 60% and FMT 70%, whose SV increased from 15.98 mL/g to 22.94 mL/g or up to 1.44 times higher than that of native starch. The increase in the FMT-modified starch could have been caused by the formation of cavities in the starch granules due to the sublimation of ice crystals in the starch granules, which allowed the starch to absorb and bind more water, leading to its expansion. Meanwhile, the increase in the SV value of FMT + ozonated-treated starch could have been caused by the formation of cavities and the increase in carbonyl (-CO) and carboxyl (-COOH) groups during the oxidation process [ process ; the sch process ,36.p < 0.05) compared to that of native starch. The largest increase occurred in adlay starch modified by FMT 60%, FMT 70%, FMT 60% + ozonation, and FMT 70% + ozonation, which exhibited an increase up to 1.44 times higher that of native starch. The increase in the solubility of FMT-modified starch and ozonation was the result of depolymerization of starch molecules, which increased along with the weakness of the granule structure due to ice crystallization in the granules, followed by sublimation. The depolymerization was also caused by the formation of carbonyl (-CO) and carboxyl (-COOH) groups during the oxidation, as shown in the schematic diagram of the oxidation of the starch reaction  compared to that of native starch, with the highest increase occurring in adlay starch modified by FMT 60% + ozonation and FMT 70% + ozonation, which exhibited an increase in solubility up to 1.26 times higher than that of native starch. The increase in the WAC in adlay starch modified by FMT and ozonation was caused by the formation of pores or cavities due to water crystallization in the granules followed by sublimation, leading to a higher starch surface area and the absorption of more water. The more porous the material, the higher the water absorption rate. The WAC in starch was also affected by the presence of hydroxyl and carboxyl groups in starch molecules. The oxidation of starch by ozonation splits the C-C bond into a carboxyl group. When the number of hydroxyl and carboxyl groups in starch molecules is large, the ability to absorb water increases [Based on ncreases ,39. The Color chromaticity is expressed in values of L*, a*, and b*, while the total color change is expressed as ΔE*. The value of L* indicates the brightness level, which has a scale of 1–100 from black to white; the value of a* indicates the presence of red if positive (+) and the presence of green if negative (−), while the value of b* indicates the color yellow if positive (+) and indicates blue if negative (−). The color of native adlay starch and adlay starch modified by FMT and ozonation can be seen in p < 0.05). The values of a* were in the range of 0.15–0.20, where the treatments of FMT 60%, FMT 70%, and FMT 70% + ozonation decreased the a* (p < 0.05). The values of b* were in the range of 3.65–4.21, where the treatments of FMT 60% increased the b* (p < 0.05). While the values of ΔE* of modified adlay starch were in the range of 0.15–0.63, indicating that the total color change was slight, all treatments increased the ΔE* (p < 0.05). Modification of FMT used a low temperature of −50 °C so as to prevent damage or coloration caused by reactions triggered by high temperatures. Meanwhile, the oxidation reaction induced by the ozonation process that occurs can actually cause some of the color-forming pigments to be oxidized before glucose units so that some of the existing compounds will be lost, and modified adlay starch was whiter, brighter, and exhibited a more intensive reduction in the intensity of the yellow color in starch [n starch .The pasting properties are expressed in the form of a visco-amylograph that can be seen in p < 0.05) compared to native adlay starch. This indicated that the granules of modified starch were more easily gelatinized due to the formation of cavities after the sublimation of ice crystals during FMT and depolymerization due to oxidiziation by ozonation treatment. Ozonation treatment also decreased the peak viscosity (p < 0.05), while other treatments had no effect on peak viscosity. However, FMT 70%, FMT 60% + ozonation, and FMT 70% + ozonation could increase the hold viscosity (p < 0.05) of the modified starch compared to native adlay starch. The highest increase in hold viscosity occurred in FMT 70% + ozonation, where the hold viscosity increased from 1322 cP to 1502.33 cP. The changes in viscosity during heating or breakdown indicated the stability of the paste during heating, where the lower the breakdown viscosity, the more stable the paste formed against heat. p < 0.05). This indicated that the starch modified by ozonation and FMT 70% + ozonation had better heat stability than the native adlay starch. This could be due to the increased regularity of the crystalline matrix due to the large amount of amylose formed during the FMT and oxidation by ozonation, and the formation of amylose complexes with other components could improve the stability of the paste during heating [Parameters of hot paste viscosity (hold viscosity) and breakdown viscosity were correlated to each other because breakdown viscosity was the deviation between peak viscosity and hold viscosity. Based on  heating .p < 0.05). This indicated that starch modified by the treatments of ozonation and FMT 70% + ozonation had higher stability when refrigerated and had a lower tendency towards retrogradation than native adlay starch. The decrease in setback viscosity was due to FMT and oxidation facilitating the depolymerization of amylose and amylopectin, which decreased the viscosity and retrogradation rate. The structural rearrangement of carbonyl and carboxyl in oxidized starch made the paste more susceptible to association tendencies during cooling and decreased the setback viscosity [Modification of adlay starch by FMT and ozonation  did not affect the cold paste viscosity . However, the treatments of ozonation and FMT 70% + ozonation caused a decrease in setback viscosity (iscosity ,44.The particle morphology of native adlay starch and adlay starch modified by FMT and ozonation can be seen in −1. The next wave peak was about 2929–2930 cm−1, indicating the presence of a C-H bond vibration. The next peak indicates the presence of a C=O double bond with a wave number of around 1640–1643 cm−1, which indicates the presence of carbonyl groups. The next peak indicates the presence of a C–O bond with a wave number of around 1304–1305 cm−1, which indicates the presence of carboxyl groups. The FTIR spectrum of native adlay starch and starch modified by FMT 70% + ozonation had the same pattern, and no new spectral peaks were found. This indicated that there was no change in or formation of new functional groups, but there was an increase in the intensity of the hydroxyl and carbonyl groups in the modified starch. The increase in the hydroxyl and carbonyl functional groups in the modified starch indicated a greater ability to absorb water, as well as confirmed the swelling volume in The functional groups of starch as seen in the FTIR-ATR spectra are presented in FTIR-ATR spectra also show the microstructure of starch, especially the crystalline and amorphous structures. The intensity ratio for wavenumbers 1045/1022 indicates a crystalline structure, while the intensity ratio for wavenumbers 1022/995 indicates an amorphous structure . Based oBased on the results and discussion, the significance of this study can be explained as follows: The treatments by FMT and ozonation could significantly improve the functional properties of adlay starch, as seen in the swelling volume, solubility, and water absorption capacity. The treatments also affected the pasting properties by increasing the stability of the starch paste during heating and cooling. The treatments can also produce porous starch granules that can potentially be used for various food, chemical, and pharmaceutical products.Modification of FMT and ozonation of adlay starch was able to increase water absorption capacity, swelling volume and solubility, and increase the number of pores and amorphous structures without the formation of new functional groups. The modified starch also showed an increase in the hot paste viscosity and a decrease in the gelatinization temperature, peak viscosity, hold viscosity, breakdown viscosity, and setback viscosity. Treatment by FMT 70% + ozonation gave a significant effect by producing modified adlay starch with a swelling volume of 21.10 mL/g, a water absorption capacity of 1.54 g/g, and a solubility of 9.20%, which was also more stable under heating conditions, had porous starch granules, and exhibited an increase in the amorphous structure, proving its ability to improve the functional and physicochemical properties of adlay starch.