WVP) increased, and water solubility (WS) also decreased with gelatin mass rising in the composites. On the other hand, opacity did not vary significantly due to the films’ cassava starch and gelatin ratio. Finally, optimizing the mechanical and water barrier properties resulted in a mass ratio of 53/47 cassava starch/gelatin as the most appropriate for their application in food packaging, indicating their usefulness in the food-packaging industry.The indiscriminate use of plastic in food packaging contributes significantly to environmental pollution, promoting the search for more eco-friendly alternatives for the food industry. This work studied five formulations (T1–T5) of biodegradable cassava starch/gelatin films. The results showed the presence of the starch/gelatin functional groups by FT-IR spectroscopy. Differential scanning calorimetry (DSC) showed a thermal reinforcement after increasing the amount of gelatin in the formulations, which increased the crystallization temperature (Tc) from 190 °C for the starch-only film (T1) to 206 °C for the film with 50/50 starch/gelatin (T3). It also exhibited a homogeneous surface morphology, as evidenced by scanning electron microscopy (SEM). However, an excess of gelatin showed low compatibility with starch in the 25/75 starch/gelatin film (T4), evidenced by the low Tc definition and very rough and fractured surface morphology. Increasing gelatin ratio also significantly increased the strain (from 2.9 ± 0.5% for T1 to 285.1 ± 10.0% for T5) while decreasing the tensile strength (from 14.6 ± 0.5 MPa for T1 to 1.5 ± 0.3 MPa for T5). Water vapor permeability ( Pseudomonas oleovorans bacteria [The food industry is one of the most significant users of single-use plastic from petroleum, contributing to solid waste . This plbacteria , chickenbacteria , soy probacteria , chickenbacteria , sunflowbacteria , spinachbacteria , corn stbacteria , among obacteria ,13.Manihot esculenta Crantz) is a low-cost tuber native to Thailand, and is used for starch production. Starch is a polymer composed mainly of amylose and amylopectin. Both components consist of chains of D-glucose residues with bonds α- that are interconnected through α--glycosidic bonds forming branches in polymers [Cassava or tapioca  makes chicken gelatin a potential component for developing new materials applicable to the food industry. Chicken gelatin contains considerably higher gel strength in Bloom value (355 ± 1.48 g) than bovine gelatin (229 ± 0.71 g). However, there are few studies related to films incorporating cassava starch with gelatin in an optimized formulation to maximize the composite’s properties ,27.Veiga-Santos et al. found that the addition of gelatin did not significantly affect the mechanical properties of tapioca starch sheets plasticized with sucrose . However−1) due to the -OH groups from starch, gelatin, and water-adsorbed molecules [olecules .−1. Additionally, stress bands associated with amide groups are overlapped by -OH stress bands. Bands at 3308 cm−1 and 2930 cm−1 for O-H and -C-H stretching vibrations are observed. At lower vibration frequencies, the bands associated with in-plane bending vibration for the CH2 group and bending of the C-OH group are also observed between 1413 and 1337 cm−1. Finally, the band corresponding to the stretching of the antisymmetric bridge of the C-O-C group rose at 1149 cm−1 [The characteristic bands of cassava starch are present in T1. The symmetrical stress band corresponding to the -C-H group is observed around 2928–2930 cm−1 associated with the -OH group. In addition, asymmetric stretching bands of the amide I group were observed between 1637–1652 cm−1, increasing intensity with higher gelatin content [−1 is shifted. Additionally, gelatin’s -C=O group (1743–1746 cm−1) increased, and the CH-O-CH2 groups (1009–1033 cm−1) decreased as the gelatin concentration within the polymeric matrix increased (formulations T2–T4). The shifting of the -OH band and the increase in the amide I band is probably due to hydrogen bonding of the C=O and N-H groups with the -OH group, which demonstrates strong bonding between starch and gelatin components. The exact position of the amide I band is determined by the most stable backbone conformation and hydrogen-bonding pattern associated with the stretching vibrations of the C=O (70–85%) and C-N (10–20%) group [−1, corresponding to the C=O group of amides-I. Likewise, the band located at 1544 cm−1 is attributed to amides-II rising from the stretching vibrations of the N-H groups and the stretching vibrations of the C-N group. Finally, the bands located at 3287 and 2928 cm−1 correspond to the stretching vibrations of the N-H group for an amide-I and the symmetric tensions of the aliphatic carbons CH2, respectively [Strong hydrogen bonds between starch and gelatin might generate smooth and homogeneous films, especially in the T3 formulation (50/50 starch/gelatin), as discussed in the morphological section. On the other hand, shifting at higher vibrational frequencies is observed for T2–T4 (starch/gelatin) films, especially for the strain band at 3295 cm content . Additio%) group . On the Thermogravimetric analysis (TGA) aims to evaluate the degradation of any material with increasing temperature. 2, CO, H2O) and carbon compounds present in the material [The second stage occurs between 150 and 270 °C, corresponding to the decomposition of organic materials of low molecular weight attributed to cassava starch ,35. In tmaterial . On the material .The low-resolution degradations observed in the DTGA of samples T4 and T5 are due to crosslinking reactions between carbonyl and amino groups, degrading at low speed and high temperatures. Consequently, the processing of these sheets has limitations related to possible uncontrolled responses at high temperatures ,37. On tDifferential scanning calorimetry (DSC) studies the phase transitions of the material as the temperature increases . DSC anaOn the other hand, higher amounts of gelatin introduce a higher degree of flexibility, which affects the observation of the well-defined endothermic melting peak of the material . For T4,In formulations T2–T4, rough surfaces with pores could be observed. The presence of starch reduced the heterogeneous and irregular appearance of the gelatin film (T5). However, with the introduction of gelatin, a roughness and heterogeneous surface are observed because of the spherical protein presence that intercalates between the polysaccharide chains, forming a rounded appearance. When the gelatin content increases, more intense molecular interaction between starch and gelatin occurs, decreasing the rounded appearance with a heterogeneous aspect.Formulation T3 (50/50 starch/gelatin) showed the slightest rough appearance, demonstrating good compatibility with this formulation. However, there were differences in the appearance of the starch/gelatin films. Apparent differences are likely due to hydrogen bonds between the components ,37. Thesp < 0.05) in the characterization of the mechanical properties between the samples are evident. These results indicate that the sample’s flexibility significantly increased with the increase in the amount of gelatin, decreasing, in turn, the stiffness of the material, as observed in the Young’s modulus values. The stiffness of the films, shown in p < 0.05) between T2, T3, and T4 formulations. Young’s modulus for the films ranged from 2.2 ± 1.1 MPa for T4 to 440.4 ± 46.1 MPa for T2, a behavior previously observed [observed ,45. Despp < 0.05) as gelatin concentration increased, from 6.2 ± 0.1 MPa for T2 (cassava starch/gelatin 75/25 wt.%) to 1.4 ± 0.2 MPa for T4 (cassava starch/gelatin 25/75 wt.%). These results agree with the significantly increased (p < 0.05) flexibility, from 4.6 ± 0.3% for T2 to 157.8 ± 16.2% for T4. From these observations, it is evident that the increase in the amount of gelatin in the composite films significantly decreases the stiffness of the composite. The presence of gelatin proteins with globular characteristics distorts the homogeneous molecular arrangement of the starch chains [On the other hand, tensile strength decreased significantly (h chains .WVP) values of cassava starch/gelatin films are described in p < 0.05) between the treatments evaluated. The treatment without cassava starch, containing only gelatin (T5), presented the lowest value, while the treatment with 100% cassava starch (T1) showed the highest value. It can be inferred that the variation in gelatin/starch concentration in the films affects water vapor permeability. The increase in cassava starch concentration significantly (p < 0.05) increased WVP values. A higher proportion of starch increases the active binding points with water molecules, favoring water vapor transport through the films. In addition, the higher the ratio of starch, the higher the proportion of plasticizer, which attracts more water molecules due to its polar character and its contribution to the formation of hydrogen bonds. These results are consistent with recent studies reporting an increase in WVP values with increasing cassava starch proportion in films obtained from chicken gelatin and cassava starch [WVP values [The water vapor permeability . An increasing trend in solubility was observed with increasing gelatin concentration in the films. The treatment without gelatin (T1) presented a significantly low value, while 100% gelatin film (T5) showed a significant increase. However, treatment T4 (75/25 gelatin/cassava starch) showed the highest solubility value, indicating that the rise in gelatin concentration increases the interactions with water, possibly due to an increase in the formation of hydrogen bridges that favor the bonding between the bioplastic and the water molecules. Likewise, the plasticizer increases with increased starch proportion, incrementing interactions with water given its polar character. However, these interactions are higher when gelatin increases in the formulation, which is associated with the creation of new hydrogen bonds.The water solubility values of the cassava starch/gelatin films are shown in The water solubility results of the cassava starch/gelatin films ranged from 3.67 to 5.15% for treatments T1 to T3, similar to those reported in a study of cassava starch and fish gelatin-based films . Treatmep > 0.05) affect the opacity values of the cassava starch/gelatin films. This indicates that the interactions between the main suspension components, cassava starch and gelatin, within the polymer matrix did not significantly affect the penetration of light through the films. The opacity values for the five treatments were between 21.20 and 22.50%. There have been previous reports with similar values for potato starch and gelatin films [The opacity values of the cassava starch/gelatin films are shown in in films , corn stin films , and casin films .Sensory characteristics play a fundamental role in the selection of packaging materials. Opacity is a property that, in addition to impacting appearance, influences the shelf life of the packaged product. For some packaged foods, films with good opacity provide high lightfastness and therefore help to improve the shelf life of photosensitive foods. However, high levels of opacity are not always required. According to the opacity values found, the films were slightly opaque due to aromatic amino acids in the gelatin acting as a barrier for light  and the WVP and water solubility (WS) values were minimized. The mechanical property values of a conventional low-density polyethylene film  were useWVP, WS, Young’s modulus, tensile strength, and deformation were 0.4 g mm/kPa h m2, 8.98%, 203.7 MPa, 3.4 MPa, and 28.4%, respectively, with predictability between 0.64 < d < 0.96. The above is obtained in films with a cassava starch/gelatin proportion of 53/47.The optimized predictions for each response variable are shown in Manihot esculenta Crantz) was purchased from Tecnas S.A. . Sodium hydroxide and glacial acetic acid were purchased from Merck . Extraction of gelatin from chickens  was reported elsewhere [Cassava starch (lsewhere . Food-grv/v) glacial acetic acid solution for three hours. Then, a heat treatment was conducted at 80 °C for eight hours in a ratio of 60 g of chicken feet in 40 mL of water. Finally, it was filtered, and the resulting solid was dehydrated in an oven at 55 °C until the material was obtained.Chicken gelatin was prepared according to the methodology previously reported , with slw/w glycerol (concerning the weight of cassava starch) to room temperature for 10 min. Subsequently, the suspension was heated to 80 °C for 15 min, cooled to 60 °C, and then we slowly added the gelatin. Finally, the rest of the suspension was sonicated at 60 °C for 50 min using an ultrasonic bath . All the blends and the simplex-lattice design are reported in Composite films’ preparation followed Podshivalov’s methodology . The casThe suspensions in plastic molds were environmentally cured for 24 h and placed for another 24 h in an oven at 40 ± 0.2 °C. After that, the solid film of cassava starch/gelatin was pilled off. The samples were placed in an airtight glass container at 50% relative humidity (RH) until the test time. The samples were cut for mechanical testing according to ASTM D6287, ASTM D618, and ASTM D882. All the thicknesses were measured in a digital micrometer. Film thicknesses ranged from 1.16 to 1.33 mm (±0.17).Thermal gravimetric analysis was run on a TA Instruments TGA Q50 V20.13 Build 39  between 30 and 700 ± 2 °C using film samples (5~10 mg) under nitrogen at a flow rate of 60 mL/min of and a heating rate of 10 °C/min. The fusion (Tm) temperatures were determined using a DSC2A-00181 system (TA Instruments) with a heating/cooling rate of 5 °C/min from −25 °C to 250 °C and cooling again to −25 °C using the differential scanning calorimetry (DSC) technique. TGA and DSC results were analyzed using the TA Instruments’ Universal Analysis software.−1 resolution and 32 scans in the range of 500 to 4000 cm−1. A diamond tip and ATR  mode were used for the test.The functional group of the films was studied using FTIR spectroscopy on an IR Affinity-1 spectrometer . The spectra were obtained using a 4 cmThe surface morphologies were scanned with an electron microscope  at 20 kV acceleration voltage, using a copper coating with the mode of secondary backscattered electrons. A gold layer was used for electron conduction on the samples.A universal SHIMADZU EZ-LZ test machine  was used for the tensile test, following the ASTM D882 standard. A 500 N load cell was used. The samples were tested with a gap of 100 mm between jaws at a speed of 50 mm/min. At least 10 samples per formulation were used. The length and width of the tested samples were 100 mm and 20 mm, respectively.WVP) determination followed the ASTM-E96. The diameter of the tested samples was 80 mm. The samples were glued to the mouth of a permeation cell containing silica gel. The permeation cells were stored inside the desiccator with 73 ± 2% RH at 25 °C. The slope of the permeation cell mass change as a function of time was obtained by linear regression. WVP (g mm/kPa h m2) was calculated as follows in Equation (1):WVTR is the water vapor transmission rate obtained from the ratio between the water gained as a function of time (g/h) and the permeation area of the sample (m2). P is the saturation vapor pressure of water (kPa); RH is the relative humidity in the desiccator, and l is the average sample thickness (m).Water vapor permeability (WS) was determined by following the method described by Cheng et al. [Wi). Then they were hydrated with 50 mL of distilled water at room temperature for 24 h with random agitation. Finally, samples were filtered and dried at 105 °C to obtain their final dry weight (Wf). WS (%) was calculated as follows in Equation (2):Solubility in water  and white patron (wP) as references. Opacity (Op) was calculated as the ratio between the opacity (bP) and opacity (wP). Both bP and wP were determined in five films with black and white standards (%) using Equation (3).The opacity test was developed using a colorimeter . Data were processed by Spectra Magic NX software. Film samples with 2 cm × 10 cm were placed under aperture for opacity measurement, determined by using black (m). The design points are given by m + 1. The five design points correspond to:A simplex-lattice design was used with two components and four lattices : ; ; ; ; . that converts the functions to a scale between 0 and 1 was used, combining them using the geometric mean and optimizing the general metric means.The optimization was performed by setting the mechanical properties of a commercial polyethylene film as a target and minimizing the values of the The experimental design and the optimization were performed using Minitab (2019) software.We used an analysis of variance (ANOVA) to establish statistical differences between treatments. Fisher’s test was used as a post-ANOVA analysis to compare the means between treatments. ANOVA and post-ANOVA were obtained using Minitab (2019) software.Cassava starch/gelatin composite films were obtained, with good mechanical properties comparable with reported values of commercial polyethylene films. The analysis of the FT-IR spectra for the composites showed the presence of hydroxyl and amide groups characteristic of starch and gelatin and with shifts of their bands at various ratios due to hydrogen bonds. Thermogravimetric analysis of the films showed that with a high amount of gelatin (T4 and T5), degradation profiles were poorly defined, characteristic of films with high flexibility. In the DSC analysis, it was evidenced that the endothermic peak Tc increased due to the presence of gelatin. Still, for T4 and T5, there was a notable decrease in its value, probably due to molecular disorder between the chains and greater flexibility.Additionally, the morphological analysis of T2 and T3 showed minor roughness, discontinuity, and heterogeneity, and a decrease in the flexibility of gelatin compared to the heterogeneous appearance of the gelatin film (T5). The results of material stiffness through Young’s modulus suggest that the flexibility of the films increased with the increase in the amount of gelatin, decreasing, in turn, the stiffness of the material . On the other hand, the water solubility results of the cassava starch/gelatin films were between 3.67 and 21.26% for treatments T1 and T5, respectively, similar to those found for cassava starch- or cornstarch-based films with gelatin.WVP, and water solubility variables were affected by the cassava starch and gelatin mixtures evaluated in the treatments. Based on the predicted values for each response in the optimization, it can be inferred that these films could be used as food-packaging material, as their mechanical properties are close to those of low-density polyethylene. Future studies could incorporate other additives to improve moisture stability properties such as WVP.With all these characterization results for the composites, an optimal formulation was obtained to develop cassava starch/gelatin-based films in a 53/47 ratio, plasticized with glycerol using the casting method, that would meet the expectations of the model polyethylene film for food-packaging applications. Young’s modulus, tensile strength, deformation, thermal,


Aspergillus fumigatus. Transmission with A. fumigatus produces aggressive aspergillosis in allogeneic haematopoietic stem cell transplant recipients, HIV patients, and cancer patients. Asthmatics and cystic fibrosis patients are allergic to A. fumigatus. MHC class-II binding epitopes can initiate immunogenic responses in patients. In this study, we deployed immunoinformatic study to reveal epitopes from fungal proteins.Among the most common causes of invasive aspergillosis and acute bronchopulmonary aspergillosis is A. fumigatus 5,8-linoleate diol synthase (ACO55067.2) and ChainB-chitinase A1 (2XVN_B). RYSYKLAGVNQVDVV, GKSFELNQAARAVTQ, and LHRYSYKLAGVNQVD epitopes interact with HLA-DRB01_0101, while ITLKLLHRYSYKLAG and KLVLRAFPNHFRGDS epitopes interact with HLA-DRB01_1501. Molecular docking analysis reveals atomic contact energy (ACE) value for these five epitopes shown below −5 Kcal/mol in docked state.In modern research, we found multiple epitopes ITLKLLHRYSYKLAG, KLVLRAFPNHFRGDS, RYSYKLAGVNQVDVV, GKSFELNQAARAVTQ, and LHRYSYKLAGVNQVD from crucial proteins of Aspergillus fumigatus. Our modern immunoinformatic research shows ITLKLLHRYSYKLAG, KLVLRAFPNHFRGDS, RYSYKLAGVNQVDVV, GKSFELNQAARAVTQ, and LHRYSYKLAGVNQVD epitopes could bind to MHC-II HLA allelic determinants and can initiate immunogenic response in patients affected by Aspergillus fumigatus.The invasive aspergillosis and acute bronchopulmonary aspergillosis are caused by harmful fungal pathogen  Aspergillus spp. Among the most common causes of invasive aspergillosis and acute bronchopulmonary aspergillosis is Aspergillus fumigatus [A. fumigatus produces aggressive aspergillosis in allogeneic hematopoietic stem cell transplant recipients, HIV patients, and cancer patients. Asthmatics and cystic fibrosis patients are allergic to A. fumigatus [Aspergillus allergens. When compared to other fungal allergens, diseases related with A. fumigatus allergens are on the rise, and it also complicates life-threatening infections in immunocompromised individuals such as cancer patients, HIV patients, and organ transplant recipients [Immunizing the immunosuppressed population vulnerable to opportunistic infections like aspergillosis may appear challenging; however, that could be considered as a first step and as the least immunosuppressed, most worthy prospects, including such granulomatous patients, living donor applicants before graft, leukaemic after effective initial treatment, solid tumour patients at diagnostic test, and healthcare workers with aspergillosis . The farumigatus . Transmiumigatus , 4. In icipients .A. fumigatus to screen out immunogenic T-cell epitopes against A. fumigatus fungi. In Fig. Only a few drugs (such as voriconazole and amphotericin B) are now available to treat this invasive condition, and even these have restrictions owing to potential risks, and so, these drugs hold longer duration of treatment with side effects , so we tA. fumigatus with various accession numbers/GenBank ID. Proteins of A. fumigatus fungi under consideration were enlisted in Table Protein database like NCBI GenBank, EMBL, and DDBJ was deployed for checking proteins of AllergenFP tool  was deplA. fumigatus via ANN algorithms. Also, affinity scores assisted in predicting binding of these epitopes to MHC-II allelic determinants. MHC-II HLA determinants were screened by using IEDB database.Epitopes were screened by using NetMHCIIpan ver.3.2 server , 10, whiAll properties of screened epitopes were determined by using ExPASy tools like ProtParam, which assisted us in finding isoelectric point (pI), GRAVY score, instability score, half-life and molecular weight, etc. .Many latest tools like I-TASSER , SWISS-PMolecular docking by using latest tools assisted us in finding binding scores, binding pocket, and H-bonds between epitopes and MHC-II HLA determinants. The latest tool PatchDock  free serDocked complexes were used to analyse undersimulation by deploying GROMACS tool , which aA. fumigatus were downloaded in fast format from GenBank-NCBI database and subjected to allergenicity analysis by using AllergenFP tool. This server produced TSI (Tanimoto similarity index) for defining resemblance of given protein sequence to reveal allergen or non-allergen nature of the given proteins  Table . InstabiStructural alphabet approach of de novo prediction was deployed to model the finalized epitopes structures. The PEP-FOLD ver.3.5 tool uses 5 to 50 amino residues for structure modelling and also performs 100 short simulations before conformation finalization for the provided sequence data, as this tool uses machine learning algorithms. Structures of epitopes modelled Fig.  were useHLA alleles HLA-DRB01:0101 and HLA-DRB01:15:01 were docked with epitopes that show interaction as per NetMHCIIpan 3.2 scores and previous screening in present research context. For molecular docking, PatchDock and FireDock tools were used. The atomic contact energy  for docked complexes was provided in Table Z-score indicates stability of structure and overall quality of the structure modelled with available datasets of X-ray and NMR models. The calculated Z-scores for complexes are as follows: (1) 1AQD-GKSFELNQAARAVTQ: −5.71; (2) 1AQD-LHRYSYKLAGVNQVD: −5.72; (3) 1AQD-RYSYKLAGVNQVDVV: −5.71; 4) 1XR9-ITLKLLHRYSYKLAG: −8.96; and (5) 1XR9-KLVLRAFPNHFRGDS: −8.96. Figure  1XR9-ITLMolProbity tool was deployed to reveal the validation of secondary structures of docked complexes by generating Ramachandran plots Fig. , and it The experimental characteristics of the dimension of sustainability and thermodynamics stages were reproduced using this force field for 100 ns. Moreover, within those sorts of simulations, where we examine behaviour at temperatures beyond 300 K, the choice of the water model is critical. TIP4P, a four-water system, was identified as the fine water model for this research. Here docked complexes were analysed for good interaction studies. An RMSD and RMSF plot clearly indicates that all the complexes hold values under 0 to 2.5 nm and 0 to 1.4 nm respectively, as provided in Fig. Aspergillus fumigatus [A. fumigatus produces aggressive aspergillosis in allogeneic hematopoietic stem cell transplant recipients, HIV patients, and cancer patients. Asthmatics and cystic fibrosis patients are allergic to A. fumigatus [A. fumigatus 5,8-linoleate diol synthase (ACO55067.2). ChainB-chitinase A1 (2XVN_B), RYSYKLAGVNQVDVV, GKSFELNQAARAVTQ, and LHRYSYKLAGVNQVD epitopes interact with HLA-DRB01_0101, while ITLKLLHRYSYKLAG and KLVLRAFPNHFRGDS epitopes interact with HLA-DRB01_1501. Molecular docking analysis reveals atomic contact energy (ACE) value for these five epitopes shown below −5 Kcal/mol in docked state. Also, docked complex was analysed for simulation analysis, and it was found that they show stable interaction pattern as per the RMSD and RMSF plots. Many previous studies show the importance of immunoinformatic study to support our analysis on fungal epitope determination likewise for Candida auris, Tropheryma whipplei [Among the most common causes of invasive aspergillosis and acute bronchopulmonary aspergillosis is umigatus . Transmiumigatus , 4. The umigatus . The immwhipplei , 22, denwhipplei , human cwhipplei , and chiwhipplei . Modern whipplei –28, althAspergillus fumigatus. Our modern immunoinformatic research shows ITLKLLHRYSYKLAG, KLVLRAFPNHFRGDS, RYSYKLAGVNQVDVV, GKSFELNQAARAVTQ, and LHRYSYKLAGVNQVD epitopes could bind to MHC-II HLA allelic determinants and can initiate immunogenic response in patients affected by Aspergillus fumigatus.The invasive aspergillosis and acute bronchopulmonary aspergillosis are caused by harmful fungal pathogen


HT reduced the contents of total starch and apparent amylose significantly in wild type but not in mutants. Meanwhile, lipids were enriched in all varieties, but the amounts of starch–lipid complexes and the RS content were only heightened in mutants under HT. HT led to greatest declines in the amount of DP 9–15 and increases in the proportion of fb3 (DP ≥37); the declines and increases were all larger in mutants, which resulted in varied starch crystallinity. The increased long-chain amylopectin and lipids may be the major contributor for the elevated RS content in mutants under HT through forming more starch–lipid complexes (RSV).High temperature (HT) during grain filling had adverse influences on starch synthesis. In this study, the influences of HT on resistant starch (RS) formation in rice were investigated. Most genes in  Oryza sativa) is an extremely important cereal and is the main food for more than 50% of the population worldwide. As the major components in rice grains, starch mainly comprises amylose and amylopectin molecules. The ratio of amylose to amylopectin and the structure of starch granules are key factors influencing rice quality. Meanwhile, amylose content and the distributions of branch lengths in both amylose and amylopectin influence starch digestibility , is not digested in the small intestine and passed into the large intestine to produce short-chain fatty acids . Due to Wx gene may regulate RS formation by tuning the amylose content   . Double  content .In a preliminary study, we developed a series of rice mutants high in RS . These mssIIIa-RNAi affected the susceptibility of grain chalky occurrence to HT exposure due to the relatively sensitive response of ADP-glucose pyrophosphorylase (AGPase) and SSI to HT exposure and induced significant decreases in AAC and a larger decline in the proportion of DP <12 under HT relative to wild type (WT). Although the formation of RS in rice grain has been investigated in detail from the intrinsic properties of starch, storage and processing conditions, and external factors such as non-starch components  distribution, X-ray diffraction (XRD) analysis and Fourier transform infrared (FTIR) spectroscopy, and thermal and pasting properties of starch were investigated. In addition, a possible mechanism of temperature affecting RS formation was proposed. The results in this study give some evidence for the impacts of temperature on RS formation in rice grain and provide alternative thoughts on improving the quality of rice high in RS under global climate warming.With global warming, high temperature (HT) during grain filling has become a great threat for rice production. HT during grain filling can lead to reduced yield, increased chalkiness, and varied amylose content (AC) and starch structure . AC has mponents , few con-1·s-1. The growth cabinets were maintained at an average air temperature of 25°C  until flowering. Then, half of the plants were transferred to another growth cabinet with an average air temperature of 30°C . Individual panicles were labeled on the day of flowering. Grains at 8 days after flowering (DAF) were collected and immediately frozen in liquid nitrogen and stored at -80°C until use. Mature grains were harvested and dried under 37°C, then the glume and aleurone were detached using Satake rice machine . After polishing, the white rice was ground into flour, passed through a 150-μm sieve and stored in a drier until use.Three indica rice varieties with different RS contents were used in this study, including R7954 (WT) and RS111 and RS4 (mutants). RS111 was derived from R7954 through 300 Gy 60-Cobalt γ-ray mutagenesis, and RS4 was obtained from RS111 by irradiation ; the twoSSIIIa was amplified using gSSIIIa primer, and the PCR products were sequenced using Genetic Analyzer . All primers used are listed in The total genomic DNA of rice leaf was extracted using Tris physiological saline (TPS)  according to the procedure modified by Actin gene (LOC_Os0350885) was used as internal reference. qPCR primers of 27 starch-related genes were referred to The total RNA from dehulled seeds at 8 DAF was extracted using Plant RNA Extraction Kit , and cDNA was synthesized using FastKing RT Kit  following the manufacturer’s protocol. Quantitative real-time PCR (qRT-PCR) was performed with SYBR Green Premix Pro Taq HS qPCR Kit  on the CFX96 Real-time system . The rice 2O, and the pH of the slurry was adjusted to neutral. Repeated centrifugation and rinsing with ddH2O were done several times until the precipitation was clearly white. Then, the obtained starch was dried at 37°C, ground, and filtered with a 150-μm sieve. The isolated starch was used for XRD, FTIR, differential scanning calorimetry (DSC), and amylopectin chain length (CL) profile analyses.Starch was isolated using NaOH as follows. The flour and NaOH solution  were mixed  thoroughly, shaking at 180 rpm and 4°C for 16 h. Then, the mixture was filtered with a 150-μm sieve, and the filtered solution was centrifuged at 5,000 g for 10 min. The yellow layer in the precipitation was scraped carefully, then the precipitation was suspended with ddHAAC and total starch content were measured according to the methods described by RS content was measured according to Amylopectin was extracted from starch using a fractionation method according to the procedure described by The X-ray powder diffractometer  was used for XRD analysis. Starch samples were packed into a round glass sample holder and scanned from 4° to 70° 2θ with a step of 0.02°/2 s. The relative crystallinity  was calculated using MDI Jade 6.5.-1, and the scanning range is from 4,000 cm-1 to 400 cm-1. The relative absorbances at 1,045, 1,022 and 995 cm-1 were extracted from the deconvoluted spectra and were measured from the baseline to the peak height.Starch granule short-range ordered structure was detected with the FTIR spectrometer . The resolution of the instrument is 4 cmΔH), onset temperature (To), peak temperature (Tp), and conclusion temperature (Tc) were calculated using Universal Analysis 2000 program (Version 4.4A).The thermal properties of starch were determined using DSC  according to the method described by The pasting properties of rice flours were measured using a rapid visco analyzer  according to the procedure described by P < 0.05 or P < 0.01) was performed using SPSS 22 .All experiments have biological triplicates. One-way analysis of variance followed by the least significant difference (LSD) test  forming crystallinity . The XRD-1 and 1,022/995 cm-1 can reflect the degree of order structure and the proportion of amorphous to ordered carbohydrate structure in starch granule external region, respectively , Tp (63.33°C–65.52°C), Tc (69.55°C–74.29°C), and ΔH (5.68–8.65 J/g) at NT, it is obvious that HT significantly elevated the gelatinization temperature (GT) and ΔH  and ΔH Table 4.ssIIIa mutants also showed very low viscosity  in all materials, but only reduced AAC and total starch content in R7954. Moreover, the varied transcripts of genes involved in starch biosynthesis under HT channeled more carbon flux to lipid synthesis, promoting lipid accumulation. The varied starch structure and lipid content under HT resulted in increased crystallinity, GT, ΔH and reduced viscosity. The high AAC and enhanced long amylopectin chains and lipid content in RS111 and RS4 under HT might boost the starch–lipid complexes further and contribute to the elevated RS.In conclusion, RS4 and RS111 are two s as b10 . MeanwhiThe original contributions presented in the study are included in the article/YZ: Investigation, Data curation, Formal analysis, Writing-original draft. ZC: Investigation, Data curation, Formal analysis, Writing-original draft. SJ: Investigation. JC: Investigation. DW: Funding acquisition, material bred. XS: Conceptualization, Formal analysis, Writing-review and editing, Supervision and Funding acquisition. All authors contributed to the article and approved the submitted version.This project was financially supported by grants from the National Key Research and Development Program of China (2021YFF100020), Functional Rice Breeding and germplasm enhancement , National Natural Science Foundation of China (32072038) and Administration Bureau of Yazhou Bay Science Technology City (SKJC-2020-02-010).The 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.


International Journal of Molecular Sciences is focused on bioactive peptides in foods or hydrolyzates of food by-products, the methods for the extraction and purification of bioactive peptides, their structural and functional characterization, and the mechanisms of action that regulate their activity and support the reported health benefits. It also contemplates those studies on the bioavailability of particular bioactive peptides using simulated gastrointestinal digestion and intestinal transport, together with the confirmation of their health benefits, as well as the recent advances and challenges in the use of proteomics and peptidomics for the analysis of bioactive peptides.This Special Issue of the Salvia hispanica L.) bioactive peptides using peptidomics, molecular simulation techniques and ensemble molecular docking analysis. The third submission, by Tonolo and co-workers [This Special Issue includes papers from nine research groups that have been invited to publish their recent advances and developments within this field. The first manuscript, authored by Martineau-Côté and co-workers , reporte-workers , deals w-workers  used hig-workers , reporte-workers  author t-workers , reporte-workers  reported-workers , is focuInternational Journal of Molecular Sciences and the scientific community.The guest editors would like to thank the contributors, peer reviewers, and journal’s managers for their kind support and contribution to this issue, and also thank the MDPI´s publishing team for expediting its handling. The guest editors hope and wish that this issue will be of interest to the readers of the


As the crosslinking degree increased, the peak strength of DMSP in starch was stronger and the specific gravity of DMSP was larger. Among the samples, LS-12CS had the highest cross-linking degree, with a greater specific gravity of DMSP. Moreover, the solubility levels of LSCSs decreased and the thermal stability and anti-digestive properties improved as the cross-linking degree increased, which was correlated with the degree of agglomeration and DMSP in LSCSs. The RS content of LS-12CS was 48.95 ± 0.86%.The structural properties and physicochemical characteristics of lotus seed cross-linked resistant starches  with different concentrations of cross-linking agents were investigated. The degrees of cross-linking of LSCSs increased along with the amount of cross-linking agent. The higher the degree of cross-linking, the greater the degree of LSCSs granule agglomeration. The occurrence of the cross-linking reaction was confirmed by the appearance of P = O at 1,250 cm Lotus seeds are an economically important specialty product in China, with a greater than 2,000-year history of cultivation. Starch, the main constituent of lotus seeds, represents approximately 50% of the dry basis and is abundant in raw material resources, especially during the processing of lotus seed milk drinks that produce numerous lotus seed starch by-products . NaturalRS4 is currently being widely studied and applied in routine food products. It is formed using chemical modification methods, such as etherification, esterification and cross-linking reactions. Cross-linking treatments are usually used in the chemical modification of starch. During the cross-linking reaction, cross-linking agents are introduced into the intermolecular bridges between biopolymer layers to undergo chemical reactions, such as esterification reaction and etherification, which may change the overall structural properties and surface chemistry of the polymer backbone . The croin vitro digestibility of LSCSs prepared by the addition of different cross-linking agents.Although much research has been conducted on the preparation and characterization of cross-linked starches, it is meaningful to study their structure-phosphorylation-function relationships in depth. No studies on the modification of lotus seed starch by cross-linking have been performed. Thus, here, lotus seed starch was reacted with different doses of cross-linking agents (STMP/STPP) in an alkaline environment to form lotus seed cross-linked resistant starches (LSCSs) with different degrees of cross-linking. The aim was to characterize their structural properties and to investigate the physicochemical properties as well as in vitro digestibility studies were provided by Shanghai Yuanye Biotechnology Co.Lotus seed starch  was isolated as described previously . Sodium 2SO4 (2 g), and the pH adjusted to 11.5 using NaOH (1 mol/L). Then, 0%, 1%, 2%, 4%, 6%, 8%, 10%, and 12% (w/w) of mixed cross-linking reagents  were added independently, and stirred at 50°C for 4 h in a water bath. Then, HCl (1 mol/L) was used to adjust the pH to 6.5, and the starch slurry was washed with distilled water until neutralization. Samples were centrifuged two to three times at 3,000 × g, then hot air-dried at 45°C for 24 h. These samples were designated as LS- CS, respectively.LSCSs was prepared using the methodology of Carmona-Garcia et al.  with sliThe total phosphorous content of each cross-linked starch was determined as previously described . The samFigure 1M1 (μg) represents the phosphorus content of the sample solution determined from the standard curve, V0 (mL) represents the volume of the sample solution, M0 (g) represents the mass of the test sample and V1 (mL) represents an equal volume of sample solution for determination.where Scanning electron micrographs of starch samples were obtained by SEM  with a magnification of 5,000× operating at an accelerating voltage of 6.0 kV, as described by Lin et al.  with somX-ray diffraction patterns and relative crystallinities of the samples were determined using an X-ray diffractometer , as described by Zhang et al. , with soAc represents the crystalline area on the X-ray diffractogram, and Aa represents the amorphous area on the X-ray diffractogram.where –1 with a resolution of 4 cm–1 and 32 scans.The FT-IR spectra of the samples were measured using an FT-IR spectrometer . The starch samples were mixed well with dried KBr at a ratio of 1:100, ground in an agate mortar under an IR lamp and pressed into round thin slices. The scanning wave number range was 400–4,000 cm31P NMR spectra were obtained using an NMR spectrometer  by solid state resonance, as described by Dong & Vasanthan  and (4), respectively.Swelling power and solubility were measured in accordance with a modified method of Miaomiao . The samW (g) represents the weight of a starch sample; and A and P(g) represent the weights of the dried supernatants and swollen granules respectivelywhere ΔH), onset temperature (To), peak temperature (Tp) and conclusion temperature (Tc), were recorded.The thermal properties of the samples were measured using a differential scanning calorimeter , as described previously . Thermalin vitro digestibility of starch was analyzed in accordance with the method of Zheng et al. .All the experiments were repeated at least three times. Experimental graphics were processed using Origin 9.0 . Data were analyzed and statistical significances were determined using DPS 9.5  (The effect of cross-linking agent (STMP/STPP) addition on the degree of cross-linking of LSCSs is shown in Scanning electron micrographs of LSCSs are shown in On the basis of the source, a natural starch crystal structure is categorized as A, B, C or V type. The characteristic diffraction peaks of the crystalline structure of A-type are at 15°, 17°, 17.8°, and 23°(2θ), and the characteristic diffraction peaks of the crystalline structure of B-type are at 5.6°, 17°, 19.5°, 22°, and 24° (2θ) . PreviouThe X-ray diffraction pattern of LSCSs is shown in The relative crystallinity of LSCSs samples are shown in –1, which was attributed to inner- and intro-hydroxyl group stretching vibration, was observed in all the starches. The band at 2,930 cm–1, which was strong and sharp, represents the anti-symmetry stretching vibration of the carbon-hydrogen bond  under alkaline conditions, the ring-opening attack of starch alcoholate ions on the cross-linker first formed mono-amyl triphosphate, which was unstable during the synthesis reaction. It had four negatively charged ionized hydrogens, and all three phosphoryl groups on its triphosphoryl chain had a strong ionizable acidic OH. When hydroxide or starch alcoholate ions react with monostarch triphosphate, pyrophosphate is more likely to detach from the group than orthophosphate, which leads to the reaction of an RO- or OH- ion with the α-phosphorous of the triphosphonyl group to form DSMP or MSMP, respectively. With a further reaction, the γ-phosphate group exfoliates, which in turn generates MSDP. Thus, when a small amount the cross-linking agent was added, the cross-linking reaction was not strong enough to generate MSDP.The LSCSs had strong signals in the range of -1 to 0 ppm and a weak signal in the range of 0 to 1 ppm, which indicated that the cross-linking reaction mainly produced DSMP. A stronger signal in the -1 to 0 ppm range occurred as the degree of cross-linking increased, leading to a greater content of DSMP. Because the hydroxyl groups at C-6 and C-3 were more reactive to the cross-linking reagent than those at C-2, DMSP may only occur within the hydroxyl groups at C-6 and C-3. Lack et al.  found th31P NMR analysis, the higher cross-linked LSCSs contained more phosphate groups forming covalent bonds, and the specific gravity of DMSP was greater. Additionally, the starch molecules were strengthened by covalent bonding, which caused a significant decrease in solubility. Shi et al.  is a measure of the integrity of the microcrystalline structure of amylopectin. The higher the values of Tc–T0, the wider the heat absorption peak , SDS (20–120 min) and RS (> 120 min) in accordance with the time of starch digestion (The starch fractions of the LSCSs are shown in igestion . The modto RDS < 0 min, SDDifferent concentrations of cross-linking agents had significant effects on the structural properties and physicochemical properties of LSCSs, and the degree of cross-linking of LSCSs increased along with the addition of cross-linking agents. The cross-linked starches all exhibited appearances similar to the original starch, but the cross-linking agent played a “bridging” role, causing the granules to gradually aggregate, which resulted in more starch chains being “bound” together by the covalent bonds formed by the phosphate groups. The covalent bonds formed by the phosphate groups in the LSCSs were mainly composed of DMSP. The level of DMSP formed was correlated with the physicochemical properties of the LSCSs. Moreover, the cross-linking reaction mainly occurred in the amorphous regions. Compared with LS-0CS, the more complex and amorphous internal structures of LSCSs allowed the cross-linking reaction to raise the transition temperature, but lowered ΔH and the solubility, which was also related to the degree of LSCSs agglomeration. The RS increased along with the level of cross-linking, indicating a limitation for the digestive enzymes. The higher resistance of LS-12CS to digestion was not only related to its crystallinity, high DSMP specific gravity and other structural properties, but also to its low solubility. Based on its low solubility, heat resistance and high RS content, LS-12CS is a potential prebiotic for the food industry.The original contributions presented in this study are included in the article/supplementary material, further inquiries can be directed to the corresponding authors.LL: writing the original draft, investigation, and methodology. SH and YL: conceptualization and software. BZ, YZ, and HZ: writing – review and editing and formal analysis. All authors contributed to the article and approved the submitted version.


Small and medium-sized enterprises (SMEs) play an enormously crucial part in the modern world economy, demonstrating the most unique and incredible ground-breaking system. SMEs’ employment statistics and future worker needs make it a focus of policies among rising economies, and Pakistan is no exception. The working conditions in SMEs diverge from industry to industry; however, irrespective of the industrial categorization, the SMEs are failing to protect the workers’ rights in the perspective of Sustainable Development Goals (SDGs). The interpretivism paradigm and purposive sampling, narrative inquiry, and analysis method have been adopted to gain in-depth knowledge of workers’ rights in SMEs concerning the SDGs. Results revealed that Pakistan-based SMEs argue to be financially weak and perceived as less equipped to adhere to the world’s standards. Highlighted issues in failure to adhere to worker’s rights include lack of financial resources, top management’s commitment, regulatory framework, SDG awareness, strategic planning, and the dire need for expert guidance and consultation in translating goals to work environments. Small and medium-sized enterprises (SMEs) are a significant engine of financial expansion and socioeconomic advancement . SMEs inThe SMEs are captivating a prominence in fulfilling the business-linked SDGs for sustainable economic growth for decent work and generating a constructive shove for an elevated worth of living, education, and health for all. These enterprises significantly contribute to poverty reduction, particularly in the rural areas for uneducated but skilled men and women. They support low-income families to run their expenditures by earning at a small level . The SDGIn Pakistan, there were 5.2 million SMEs reported last year in the press . These SPakistan has been a signatory of SDG 2030 from its beginning in 2015 . The NatA similar study by In this study, we explore the failure of SMEs Management to fill the worker’s rights as required by SDG 2030. According to the International Labor Organization (ILO) conventions illustrated, workers’ rights research defines it by using the terms “freedom of association, collective bargaining, the abolition of forced or compulsory labor, equality of opportunity and treatment in employment and occupation, protection of children and young persons, wages, working hours, safety, and the legal employment relationship.” We have studied SMEs’ workers’ rights in Pakistan following the SDGs agenda 2030. We explore among the 17 goals directly related to workers’ rights, and SMEs can contribute to it. We studied why SME management has failed to fulfill the workers’ rights required by these related SDGs. How these workers’ rights can be secured; to what extent the government may support SMEs; and how can these SMEs support Pakistan’s national economic growth?The SMEs are enterprises with low budgets that contribute to the country’s economy. There appears to be no universally accepted definition of SMEs. The SMEs definitions are still confusing concepts as they vary. For example, SMEs have less than 500 workers in France. The description may vary according to the kind of business in Japan, where mining, manufacturing, shipping, and building industries are defined as SMEs with less than 300 workers. In Ghana, SMEs are defined as an enterprise with an annual income of “23,700–2,370,000” dollars . In ThaiNo standardized description of SMEs exists in Pakistan . The “SMSmall and medium-sized enterprises (SMEs) play a significant role in developing a country’s economic, industrial, and social domains. For this study, we have divided SDG into three groups. In Group 1, all the goals directly related to workers’ rights in SMEs are included. Group 2 consists of the SDGs that are related to the environment. In Group 3, only SDGs associated with the ideal layer are included in Group 1.SDG 1: Poverty reduction in all its forms and domains.SDG 2: Hunger reduction, food security, and promote sustainable agriculture.SDG 3: Guarantee health services and endorse wellbeing plans for all age groups.SDG 4: Ensure a wide-ranging and reasonable value-added education program and uphold lifetime opportunity for all with no discrimination.SDG 5: Gender equality and women and girls empowerment.SDG 8: Promote sustained, broader and sustainable economic growth and productive employment and decent workplace.SDG 10: Condense inequalities between inter and intra nations.SDG 16: Promote peaceful and inclusive societies for sustainable development, provide access for all and build effective, accountable, and inclusive institutions at all levels.SDG 17: Strengthen the means of implementation and revitalize the global partnership for sustainable development.SDG 6: Ensure availability and sustainable management of water and sanitation for all.SDG 7: Ensure access to affordable, reliable, sustainable and modern energy to all.SDG 9: Build resilient infrastructure, promote inclusive and sustainable industrialization, and foster innovation.SDG 13: Take urgent actions to combat climate change and its impact.SDG 14: Conserve and sustainably use the oceans, seas, and marine resources for sustainable development.SDG 15: Protect, restore, and promote sustainable use of terrestrial ecosystems, sustainably managed forests, combat desertification, halt and reverse land degradation, and halt biodiversity loss.SDG 11: Sustainable city and community.SDG 12: Insurance of sustainable consumption and production patterns.Our primary focus in this study is the goals given in Group 1. The workers’ rights studied concerning the objectives given in Group 1 include basic needs, health, education, income and work, peace and justice, social parity, gender egalitarianism, and networks.via phone and physically. After collecting data, narrative analysis was used to analyze it (Following the interpretivism paradigm, this research used a narrative analysis approach. As the study explores the SDGs in 2030, the employees’ perspective is chosen for analysis. It is essential to understand the individual and his perception as everyone sees the world their way. This method provides a greater understanding of the human condition and perceptions. The concept of interpretivism sheds light on the human nature that reality is viewed and interpreted according to the brain’s conditioning, which comes from experience. It provides a perspective of the situation of SGDs in the future in the interviewees’ minds and shares their understanding. A qualitative approach has been adopted to gain in-depth knowledge of SME workers’ rights concerning the SDGs described in Group 1. The purposive sampling technique is used to select the respondents, as alyze it . The samThe narrative analysis of this study showed that there are different reasons why SMEs cannot provide the workers and rights as required by SDG 2030. These reasons can be categorized into five groups based on the interviews conducted with the management of different SMEs. These categories are lack of finance, lack of top management commitment, lack of regulatory framework, lack of SDGs knowledge, and lack of strategic planning for SDGs.Most of the respondents shared that lack of financial resources is the primary reason for not providing the workers’ rights as required by SDG 2030. The majority of interviewees shared that they are willing to provide all the rights to their workers as required by SDGs, but due to the limited financial resources, they are unable to fulfill this requirement:“Being an export-oriented SME, we have different challenges. Our operations are order-based, and price competition is very high, so we find it difficult to provide all the rights to the workers. Providing each facility involves cost and requires funds, so due to the availability of limited funds, we have to remain cost-effective.” SME 1“We know that our workforce is the backbone of our business, and Sialkot has a unique type of workmanship. All our workers are earning enough to meet their personal and family needs. We have the piece-rate workers, which means more work we have more they do and make more. However, a small organization can’t afford to fulfill all their rights due to the non-availability of funds as our profit margins are meager.” SME 7“It seems very hard to provide all the workers’ rights required by the International Standards. Registration with Social Security, EOBI, paying them for annual, sick, casual and maternity leaves, paying the overtime on premium rates and providing them health and safety facilities all need money beyond the affordability of a small organization.” SME 7The respondents expressed that if they have enough financial resources, they will provide all the rights to their workers and try to perform their role to achieve the SGDs 2030. They believe that they will be more productive if the workers are facilitated. Although the SMEs enable their workers, they can do more for their workers if they have funds.Another reason for failure in providing the workers’ rights as required by SGDs 2030 is a lack of top management commitment , as poin“The lack of top management commitment in providing workers’ rights as required by the GSDs is pronounced. For them, it is on least priority. Top management focused on business expansion, building expansion, and purchasing new lands. They are not interested in solving the issues workers face in their lives. They are not ready to give workers a permanent job and prefer to hire them on piece rate bases.” SME 2“The top management is unwilling to take any step to improve workers’ lives. Even the working environment in the organization is not a decent working place and has many health and safety risks, and the top management does not have any plan to improve it. They are not willing to eliminate the discrimination especially based on gender. They are not willing to participate in any collaborative program to enhance the capacity building of their workers through training.” SME 5As we have some International certifications and our international brand is very conscious about the workers’ rights, our top management provides all the necessary resources for maintaining these certifications. However, there are primarily many issues related to worker rights, which have never been a priority by top management. Many things are good, but these are in the paper.” SME 8“The respondents believe that the lack of top management commitment toward the workers’ rights according to SDGs is one reason for the failure to provide these rights. Although they are trying to provide some basic facilities to the workers, it requires more attention and support from the top management to fulfill their rights according to SDGs.The interviewees also narrated that due to poor implementation of the laws related to the workers and the ineffectiveness of the government regulatory bodies responsible for protecting the workers’ rights, the workers are facing many difficulties:Although labor laws are available and the government has announced the labor policy to protect the workers’ rights, which are very comprehensive and admirable. However, the implementation of these rules and regulations is fragile. Most workers do not have job security; they do not have a job contract from the employer and can be fired without fulfilling legal requirements.” SME 9“Health and safety conditions in factories are worse, cleanliness, First aid arrangements. Fire fighting arrangements and emergency exit paths are not properly maintained. Portable water, sanitation facilities, and food serving arrangements are not per legal requirements. Although there are many departments the government has established for monitoring the availability of these facilities according to law, it seems that this monitoring is not effective.” SME 13“The government should improve its mechanism to implement the regulations already developed to protect workers’ rights. Some effective channels of monitoring and motivation should be formed. According to the SDS, organizations that provide good facilities to their workers should be given some incentives and rewards.The participants admitted that awareness of the SDGs is rare in Sialkot. On the subject, they have not heard or learned much about this. People have a scarce understanding of the SDGs 2030 and how they can contribute their roles to the achievement of these goals:“We are working on the different International Standard related to the environment, quality, health and safety, and social accountability. We have participated in different training and seminars related to these standards; however, any seminar or training was rarely held on SDGs 2030.” SME 12The SMEs do not sufficiently understand the details about the 17 SDGs in Sialkot, which are related to workers’ rights and the details about these are mostly unknown.” SME 4“It was narrated that although the regulations related to labor rights are available, their implementation is not adequate. It is difficult for the workers to get their rights to the existing framework. The role of the government in monitoring regulator authorities related to workers’ rights should be practical and supportive for implementing existing regulations. According to the SDS, organizations that provide good facilities to their workers should be given some incentives and rewards.The participants described that their business plans do not include any separate strategic planning for SDGs about workers’ rights. Being small-sized organizations, we are mainly occupied with our day-to-day operations:“SDGs related Planning is related to the Government or the big Organizations with a big workforce and enough resources. In small organizations, we don’t think that we can contribute to these SDGs, so we do not have a strategic plan for SDGs.” SME 12“Workers are our main strength and we try our best to facilitate as much as we can; however, having some sort of separate strategic planning for SDGs related to workers’ rights is not possible.” SME 15In SMEs, there are no well-described business plans. They rarely have the strategic planning for the SDGs related to workers. Due to limited knowledge about the SDGs, they do not prepare any strategic goals related to these SDGs.The interviewed persons expressed that they are competent in their core operations, and it is not easy for them to understand the SDGs and how to implement these in their current business setup:“For implementing such International requirements like SDGs, we need some experts to help us. Normally we hire some consultant; however, it is difficult to understand for our existing workforce and management staff, and we need a budget to pay for the consultant.” SME 13“SDGs related to the workers demand a lot beyond our capacity. It is not easy that we can understand them and implement these requirements with any external support. It is complex and requires a lot of effort.” SME 15The respondent narrated the complexity of the implementation of SDGs related to workers’ rights. It is required to hire some professional services to help them perform in their organization. Also, it is necessary to enhance the understanding of their existing staff toward these SDGs.Small and medium-sized enterprises contribute significantly to private sector entities in developed and developing economies. The vital role of SMEs for extensive social-economic growth like employment chances gives them precedence for attaining the SDGs. These goals are sweeping arrangements calling for the functioning of all economies to perk up the populace everywhere. SDGs aim to reduce poverty, decrease inequality, and deal with climate change. SMEs in Pakistan work on a low budget and hardly afford sustainable development agenda in their workplace for laborers. Still, despite size and income, all shareholders, employees, and customers expect a similar pledge to sustainability from all small and medium businesses . Being sBased on the research and interviews conducted by the SMEs or top management owners, some gaps in the policymaking and implementation process must be addressed as a top priority. A few policy recommendations for the adoption of SDGs in SMEs are given as follows:•The SMEs must adopt a long-term strategy for the implementation of SDGs. If SMEs adopt these goals, they will get an extensive customer range.•The SMEs owners must follow their sustainability act through sustainability exposure such as an annual report, sustainability reports, and SDGs as a stencil for reporting.•Government must encourage SMEs and facilitate them in adopting SDGs through some subsidies offers or financial assistance.•The training programs for owners of SMEs must start nationally and internationally. This will encourage them and make them aware of the significance of SDGs adoption in their enterprises.•The employees must also offer training and workshop for knowledge about these sustainability parameters.The data collected for this study was through interviews due to limited sample size. Few interviews were conducted simultaneously, i.e., cross-sectional, so the future research should expand to other cities of Pakistan with greater sample size and choose longitudinal research design to increase the generalizability of results for other countries. Purposive sampling has been used to fulfill the study’s need to seek the narrative of respondents, so the sample chosen was based on their knowledge of SDGs. Future researchers should use a different sampling technique, such as cluster sampling, to get a more significant portion of the population to offer their insight on the matter.A conceptual model can be incorporated in the future for sustainable development as the awareness of SGDs in developing countries is low. The model may include SME performance, SDGs awareness, and financial assistance. Other factors can be added depending on the economic conditions of the country. Through research, the knowledge gap can be overcome and create awareness among workers. When the SMEs are self-sustainable, the poverty levels of society can be reduced, and necessities can be accessed more easily than before. With passing the time, natural resources are depleting, due to which SMEs need to be self-sustained. To do so, the UN has provided a future guideline for SDGs compliance to manage the organization in these challenging times. Managers can learn from this study and accordingly meet the requirements of SDGs to make sustainable decisions.The original contributions presented in this study are included in the article/supplementary material, further inquiries can be directed to the corresponding author.RN proofread the first draft composed by MR. MK made some improvements to the draft. SRS and SS helped with the data collection. AA refined the statistical analysis. HI improved the draft to be ready for publication. All authors contributed to the article and approved the submitted version.


The physical, mechanical, and thermal properties of the films were evaluated. The use of ANN starch decreased the solubility (from 21.8% to 19.5%) and the swelling power (from 299% to 153%) of the film samples. In addition, an increase in opacity and relative crystallinity (from 7.54% to 10.5%) were observed. Regarding the thermal properties, all the samples presented high stability to degradation, with degradation temperatures above 200 °C. However, the samples showed deficiencies in their morphology, which affected the barrier properties. The use of ANN starch has some advantages over native starch in preparing films. However, more analysis is needed to improve the barrier properties of the materials. This work reveals the potential of the ANN ulluco starch for biodegradable film preparation. In addition, the use of modified ulluco starch is an alternative to add value to the crop, as well as to replace non-biodegradable materials used in the preparation of packaging.This work aimed to evaluate the use of annealing (ANN) ulluco starch in the preparation of biodegradable films and its impact on the physicochemical properties of the materials. Three film samples  were prepared at a fixed starch concentration (2.6%  Plastics are inexpensive materials with exceptional characteristics, such as great strength, versatility, water impermeability, low conductivity, corrosion resistance, and durability, making them an essential raw material for different industries. As a consequence of its industrial importance, in 2020, the worldwide production of plastic reached 367 million metric tons . ProductTg) and the onset of gelatinization (To) and with an excess of moisture  along with its amylose content (between 28% and 36% d.b.), which makes it suitable for use in the production of biodegradable packaging due to the high transparency, excellent stability against temperature, and good texture and water vapor permeability characteristics [In recent years, starch obtained from ulluco  were acquired at a local market in Ibagué (Colombia). The native starch (amylose content 27.9%) and the annealing modified starch (ANNS) were obtained according to the methodology described by Valcárcel-Yamani [Ulluco tubers (l-Yamani  and modiw/v), with magnetic stirring for 5 min. The mixture was heated to 95 °C and maintained at this temperature for 5 min. After that, the mixture was cooled to 50 °C, and glycerol was added at a concentration of 1.0% (w/v). The solutions were stirred for 10 min and then transferred to a Petri dish with a diameter of 8.5 cm (~0.53 g of the film solution per cm2). The films were dried at 40 °C for 24 h. After the drying process, native ulluco starch film (FSC), ANNS1 film (FS1), ANNS2 film (FS2), and ANNS3 film (FS3) were obtained and stabilized at a relative humidity (RH) of 57 ± 3% and a room temperature (~25 ± 2 °C) for at least 24 h, before the analyses.The films were prepared using the casting method following the methodology described by . Native Micrographs of the surface of the samples were acquired using scanning electron microscopy  at an accelerating voltage of 15 kV under vacuum pressure of 40 Pa and different magnifications. For the analysis, samples were placed on stubs using double-sided carbon tape and covered with a gold layer.Ac) and the total area (At) using Equation (1) [X-ray diffraction patterns were analyzed using a Bruker D8 Advance diffractometer  operating with Nickel filtered and Cu-Kα1 radiation, 40 kV and 40 mA. The wavelength used was 1.5406 Å. Data collection was carried out in the 2θ range of 3.5–70°, with a step size of 0.02035° (2θ) and a counting time of 0.8 s/step. Relative crystallinity was determined after deconvolution of peaks from diffractograms by the relationship between the crystalline peak area (tion (1) . ResultsSwelling power (SP) and solubility (S) were calculated using the methodology reported by Baron et al.  with somThe water vapor permeability (WVP) was determined according to . ResultsThe absorbance of US edible films was measured at 600 nm using an ultraviolet–visible spectrophotometer . The OP was calculated using Equation (4).Samples with dimensions of 5.0 × 1.0 cm were loaded in a texturometer  and their mechanical properties were evaluated according to . The cro−1 and 40 scans over the range of 4000–700 cm−1.The Fourier-transform infrared spectroscopy (ATR-FTIR) spectra of samples were determined using a Fourier transform infrared spectroscopy  equipped with the Platinum ATR accessory (Bruker). The spectra were obtained with a resolution of 4 cmThe weight loss (WL) and thermal degradation temperature (TD) of film samples were assessed using a thermogravimetric analyzer  according to the method described by . A tempeThe dynamic-mechanical thermal behavior of film samples was evaluated using a dynamic mechanical analyzer  following the method described by  with som® software .The mean and standard deviation of the results for at least three replicates under each treatment were reported. Analysis of variance (ANOVA) at the 95% confidence level was used to determine any statistically significant differences. Multiple range tests (MRT), using the Tukey–Kramer range test, were used to determine which mean results differed significantly from others. ANOVA and MRT were performed using MATLABMicrographs of the films evaluated are shown in p < 0.05). The difference between the relative crystallinity values of the films can be associated with the reorganization of the starch granule structure in the modification process [The XRD patterns of biodegradable films are presented in  process . Other r process . High rep < 0.05). The decrease in these parameters may be because the ANN process produces a reorganization of the molecular structures in the starch granule, strengthening the interactions between amylose and amylopectin, thus generating a more stable structure, or increasing the crystallinity of the starch [The solubility and swelling ability of the films obtained with annealed starch were lower than the FSC . Howevere starch . Since se starch  and potae starch  but lowee starch . Despitep < 0.05), followed by FS2, FS1, and FS3. A decrease in the swelling power was observed in the films prepared with ANN starch compared to FSC. Such decreases in swelling power in films prepared with modified starch are related to the reduction of free volume in the film structure, which is a consequence of the crystallinity increase [The FSC showed the highest swelling power (increase . Althougincrease . As mentp < 0.05). The differences in permeability in the analyzed samples can be related to the defects (macrocracks) in the morphologies of the films obtained with ANN starch. An increase in permeability related to defects in morphology was also observed in films prepared using a mixture of hydroxypropyl methylcellulose/hydroxypropyl starch [The purpose of food packaging materials is to isolate the food matrix from environmental factors such as light, oxygen exposure, and water vapor, thus increasing its shelf life. The permeability of a film is defined as the amount of water vapor that passes through a unit area of the material per unit of time . The WVPl starch . In addil starch .p < 0.05), while FS2 and FSC presented low opacity. A similar increase in the opacity of films obtained with HMT-modified rice starch was observed compared to those obtained with native starch [Transparency is a parameter that can help define the final use of food packaging since it is related to sensory perception by the customer as well as product protection. The opacity of biodegradable films is shown in e starch . The autp < 0.05). The higher value of EB can be related to the low relative crystallinity of the sample  [−1 [−1 and 2950 cm−1 [The ATR-FTIR spectra of samples analyzed are shown in s (–O–H) . A previO–H) [−1 . HoweverO–H) [−1 . It may O–H) [−1 . The asy950 cm−1 .-1 and 1430 cm-1 were observed, corresponding to the bending vibration of −CH groups [−1 and 1078 cm−1 are related to the crystalline structure, whereas the band observed at 997 cm-1 is associated with the amorphous region correlated to the presence of starch in the film composition [Additionally, bands at 1320 cmH groups . The banp > 0.05). The samples showed four stages of decomposition (The weight loss of films was 86.9 ± 1.9%, 88.8 ± 0.7%, 86.6 ± 1.1%, and 86.3 ± 0.5% for FSC, FS1, FS2, and FS3, respectively (position : The firUlluco starch modified by ANN showed potential as a material to produce biodegradable films. However, a significant influence of the modification process on the physicochemical characteristics of the materials evaluated, mainly in the morphology, was evidenced. In general, the modified ulluco starch improved film characteristics such as solubility and swelling power, providing greater resistance to moisture. In addition, an increase in relative crystallinity was observed, associated with a decrease in the transparency of the films. This fact may contribute to the consideration that a reduction of the transparency of the films could be related to a lower passage of light and greater protection of the food matrix. Although an increase in the crystallinity of the samples is observed, there is no evidence of a relationship between this characteristic and the barrier properties of the materials, which could be influenced by the micro and macrocracks observed on the surface of the samples. This observation suggests that more tests should be carried out to determine the optimal concentration of modified starch and plasticizer in the preparation of biodegradable films as well as the starch gelatinization time and thus obtain continuous and more consistent morphologies.On the other hand, all the samples presented high thermal stability. However, no impact on this property was observed using modified starch. In general, ulluco starch modified by ANN offers as a viable alternative for producing biodegradable films with improved physicochemical properties. However, additional testing is required to optimize its performance.


Extrusion is a method of processing that changes the physicochemical and rheological properties of starch and protein under specific temperature and pressure conditions. In this study, twin-screw extrusion technology was employed to prepare yam starch–soy protein isolate complexes. The structure and properties of the complexes and their effects on the quality of dough were studied. The results showed changes in the X-ray diffraction, rheology, and in vitro digestibility of the complexes. The extruded starch–protein complex formed an A+V-type crystal structure with the addition of soy protein isolate. A small amount of soy protein isolate could improve the complex’s viscoelasticity. As the content of soy protein isolate increased, the content of slow-digesting starch and resistant starch in the complexes increased, and the digestibility decreased. The microstructure of the dough indicated that the network structure of the puffed yam starch–protein complex dough was more uniform than that of the same amount of puffed yam starch. The moisture distribution of the dough showed that with the addition of extruded flour, the closely bound water content of the dough increased, and the weakly bound water content decreased. The hardness, gumminess, chewiness, and resilience of the dough decreased. In conclusion, extruded starch–protein complexes can improve dough quality and provide technical support for the broad application of yam. Starch and protein can regulate the function of the body and play very important roles in the body, with great research potential. In addition to their nutritional properties, starch and protein can also be used as gelling agents, thickeners, and stabilizers in food systems. Yam starch is used not only in the field of food but also in the field of medicine. It greatly affects the physicochemical properties of food, such as the texture and rheology . The proThe high temperature, high pressure, and high shear force generated in the extrusion process can promote the movement, migration, and extension of the starch molecular chain structure, aggregated structure, and protein molecules. Thus, extrusion has the potential to enhance the interaction between starch and protein molecules . During The dough is prepared by mixing flour with water, and different ways of treating the flour can affect the properties of the dough. The stability of dough has been altered by the addition of different varieties of starch or the addition of treated modified starches to wheat dough. For example, dough prepared by replacing part of the wheat flour with gelatinized corn starch had an increased apparent viscosity , and theIn this study, twin-screw extrusion technology was employed to prepare yam starch–soy protein isolate complexes. The structure and properties of the complexes and their effects on the quality of dough were studied. This experiment establishes a foundation for future research on the effects of soybean protein isolate on the properties of yam starch during extrusion processing, encourages the full utilization of yam resources in wheat dough, and provides new ideas for further preparations of bread, noodles, etc.w/w, dry basis) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. . Pancreatin  and amyloglucosidase  were obtained from the Sigma Corporation . The glucose oxidase peroxidase (GOPOD) glucose kit was purchased from Megazyme Co., Ltd. . All other chemicals used in this study were of analytical grade.Yam starch (YS) was extracted from Chinese yam  in our laboratory at the College of Food and Bioengineering, Zhengzhou University of Light Industry. According to the determination method of Ali et al. , the meaw/w of starch). The YS/SPI mixture was extruded using a parallel twin-screw extruder  with an 11 mm barrel diameter and 820 mm screw length. The extrusion processing was carried out at 120 °C with the temperatures of the different barrel zones set as 50, 60, 80, 80, 80, 130, and 130 °C. The material moisture content and screw speeds were 30% and 180 rpm, respectively. The extruded product was cut manually and placed in a drying oven at 40 °C for 24 h and then ground and passed through a 149 micro-meter sieve. Based on the added amount of SPI, the extruded YS-SPI complexes were labeled as EYS , SPI-5%, SPI-10%, SPI-15%, and SPI-20%. The extruded YS-SPI complexes were prepared using the method of Zhang et al. with minor modifications . Yam staAn X-ray diffractogram of the extruded product was obtained using an analytical diffractometer . The diffraction patterns were recorded in the range of 5~35° (2θ). The scanning speed was 2°/min and the scanning step was 0.02°. The software Jade 6.0  was utilized to calculate the degree of relative crystallinity (RC) of the sample .The endothermic flow curves of the YS-SPI complexes were obtained using a differential scanning calorimeter . Baseline calibration and oven temperature calibration were performed with empty aluminum dishes and indium prior to the experiment. An empty pan was used as the reference. The samples  and distilled water were added to the aluminum pot using a microliter syringe for a total weight of 12 mg. The samples were equilibrated at room temperature for 12 h and then gradually scanned in a temperate range from 20 °C to 120 °C at a heating rate of 10 °C/min. The resulting data were processed with TA 2000 analysis software.The dynamic rheological behavior of the starch–protein complex was measured using a discovery rheometer . The sample was equilibrated at 25 °C for 5 min before measurement. The sample preparation referred to the method of Shi et al. , with apThe in vitro digestibility of the samples was determined according to the slightly modified method of Englyst . The sam20 is the glucose content after 20 min of hydrolysis; G120 is the glucose content after 120 min of hydrolysis; and FG is the free glucose content of the sample.Based on the absorbance of the sample and standard glucose, the contents of fast-digesting starch (RDS), slow-digesting starch (SDS), and resistant starch (RS) were calculated, respectively, and the relevant formulae were as follows:w/w) of WF to obtain the dough samples. Dough made from natural wheat flour (WF) was used as a control. Based on the substitution quantity of wheat flour, the dough samples were labeled as EYS-10%, EYS-20%, SPI-10%-10%, and SPI-10%-20%.The EYS and SPI-10% were used to replace 10% and 20%  for 1 d and then cut into cubes of approximately 10 × 4 × 5 mm, and the microstructure of the samples was observed using a scanning electron microscope  at an accelerating voltage of 10 kV. The tested sample was placed on the sample stage with black conductive adhesive, gold-sprayed by vacuum sputtering, observed, and photographed for preservation.According to the assay method of Wang , implemeThe textural analysis was performed according to the method of Shi , with mip < 0.05) were performed using the SPSS 26.0 statistical software program to determine significant differences between the means for each metric.All experiments were repeated three times, and the mean and standard deviation were calculated. After sorting and measuring the data using charts, Origin 9.0 software was used for the analysis. Analysis of variance (ANOVA) and post hoc Duncan’s multiple range test  of the yam starch changed slightly with the addition of the soy protein isolate. It may be that the integrity of the starch granules was more severely disrupted by the addition of soybean isolate and extrusion, and the interaction between the amylose and amylose or amylopectin was weakened, which made the yam starch easier to paste. Compared with the extruded yam starch, the gelatinization enthalpy value (ΔH1) of the starch in the extruded complexes decreased with the increase in the protein addition ratio. When 20% soy protein isolate was added, the ΔH1 decreased from 1.87 J/g to 1.26 J/g. The presence of protein reduces the interaction between starch and water in the complex, because it affects the water mobility of starch granules during gelatinization [2 value.The gelatinization parameters of the YS-SPI complexes are summarized in nization . TherefoThe dynamic rheological curves of the yam starch with different amounts of soy protein isolate are shown in As can be seen in The tan δ value is the ratio of G” and G’. The larger the value of tan δ is, the stronger the system’s fluidity is. The smaller the value of tan δ is, the stronger the solid performance is. The presence of SPI increased the tan δ value of the complex, indicating a reduction in the solid-like behavior of the system. With the increase in the soy protein isolate concentration, the viscoelastic behavior transformed from a solid-like state into a liquid-like state .The digestibility of the samples was calculated by measuring the glucose released during starch hydrolysis. Studies have shown that the digestion degree and speed of starch and starch-containing foods are related to the release of glucose, which is represented by the contents of RDS, SDS, and RS in the starch system .p > 0.05). Compared with the extruded YS, the SDS and RS contents of the extruded YS-SPI complexes were significantly increased, and the RDS content was significantly reduced. When the content of soy protein isolate was 20%, the RDS content decreased to 76.86%, and the RS content increased to 19.93%. This indicated that the digestibility of the complex gradually reduced with the increase in the proportion of soy protein isolate. This may be related to the protein matrix enwrapping the starch networks and restricting hydrolysis. The proteins were enwrapped and surrounded by starch granules, which may have been a barrier to the delay of enzymatic hydrolysis [As shown in drolysis . Ezeogu drolysis  found thdrolysis  also fouUnder the same conditions, the network structure of the dough with the same amount of extruded YS-SPI complex was more uniform than that with extruded YS. During extrusion, the high temperature caused the irreversible denaturation of the soy protein isolate. Under conditions of adequate moisture, the increase in the temperature led to the expansion of the protein structure and the exposure of hydrophobic groups. During subsequent aggregation, the denatured proteins were stabilized by hydrophobic interactions and disulfide bond formation .21 (0.01~1.00 ms) represents the bound water closely bound to starch and gluten proteins. T22 (1.00~50.00 ms) represents the weakly bound water bound to macromolecules such as starch and protein. T23 (50.00~200.00 ms) represents free water. A21 represents the peak area percentage of the bound water. A22 represents the peak area percentage of the weakly bound water. A23 represents the peak area percentage of the free water [The distribution of moisture in the dough was measured using low-field NMR. The parameters related to the moisture distribution of the dough are shown in ee water ,34.22 (weakly bound water) > A21 (bound water) > A23 (free water). With the addition of extruded flour, the proportion of closely bound water A21 increased significantly, and the proportion of weakly bound water A22 decreased. The higher the ratio of A21 is, the higher the bound water content of the dough will be. As a result, the better the dough’s water-holding capacity is, the more stable the dough’s structure will be [23 decreased as the additional amount increased, and the dough’s fluidity weakened. This showed that the extruded flour’s hydrophilic effect reduced the amount of free water in the dough and increased the ability of the water molecules to be bound. At this time, the T21 of the bound water in the dough due to its combination with the extruded flour was correspondingly increased. At the same substitution amount, the free water in the dough with the extruded starch–protein complex added was lower than that to which extruded yam flour and starch were added. This can be related to starch–protein interactions during extrusion. The results indicated that the gelatinized starch and denatured protein in extruded flour have sufficient water binding sites and can also affect the movement of water molecules. Thereby, the moisture distribution of the dough was promoted.The proportions of the three states of the water in the dough were as follows: A will be . The freThe texture analysis of dough is an indispensable theoretical method for the study of dough. The internal tissue state of the dough can be objectively analyzed using a texture analyzer. According to Under the same conditions, the dough with extruded YS-SPI complexes had a higher hardness, springiness, gumminess, and chewiness than the dough with extruded starch. This may be due to the attachment of the soy protein isolate molecules to the surface of the starch granules, which affected the water absorption and expansion of the starch granules, and the exudation of amylose molecules. The resilience of the dough decreased with the addition of the extrusion flour. Additionally, it decreased with the increase in the replacement amount of extruded flour. A study conducted by Abdelghafor et al.  showed tIn this paper, the structure and physicochemical properties of the extruded YS-SPI complexes and their effects on dough quality were investigated. The extruded YS-SPI complexes formed an A+V-type crystal structure, and the starch digestibility was reduced. The low content of soy protein isolate could also improve the viscoelasticity of the complex. The tightly bound water content of the dough increased. The fluidity of the dough was weakened. Under the same conditions, the hardness, springiness, gumminess, and chewiness of the dough with the extruded YS-SPI complexes were higher than those of the extruded YS. In conclusion, the extruded YS-SPI complexes not only added nutrients to the food but also improved the quality of the dough. Therefore, dough can be modified by adding extruded flour, providing a new green and pollution-free technical idea for the production of fried dough sticks, dried noodles, bread, and other products.