Starch granules and other plant tissues are commonly found as part of the microdebris assemblage analysed within dental calculus. These are often interpreted as evidence of past diets. However, many of the starch granules extracted from dental calculus are intact, and do not show evidence of alterations as a result of being processed for consumption. This research examines if plant material can accidently enter the mouth while being processed for a meal, with a focus on starch granules. Grinding experiments were performed on three types of cereal grains . We compare the presence of intact and altered starch granules in mouthwash samples  from individuals involved in grinding and also from samples in the environment surrounding the grinding activity. This experiment is a proof of concept aimed to expand experimental research in the field of dental calculus analysis and to encourage the exploration of pathways beyond direct and deliberate consumption. Starch granules entrapped within human dental calculus can provide valuable information on past diet e.g. , particuOryza spp.) and millets  in Bronze age samples from Shilinggang, southwestern China, to pulses (Fabaceae) and cereals (Triticeae) from medieval Leicester, England (Ipomoea batatas) in Rapa Nui, Easter Island, dating prior to European contact , wheat  and common oat . These cereals were selected for several reasons. Firstly, the starch granules produced by these crops are morphologically and diagnostically different: wheat starch is composed of single oval to round starch granules , with a bimodal distribution, and are easily contrasted in modern reference material with those from oat (compound starch granules) and those in millet (single starch granules). This enabled the counts of starch granules produced during each experiment to be as accurate as possible. Secondly, using these different grains enabled us to assess if the starch morphology of crops influences their airborne properties. For example, starch granules from oats are known as compound granules because they tend to be in composite particles made up of multiple granules, which might lead to them being less airborne than single starch granules of wheat. Thirdly, these crops played a significant role in the diet of human societies in the past. For example, wheat and oat were dietary staples in many parts of Medieval Europe  took place at a ‘Viking’ domestic house structure at Murton Park, Museum of Farming, Yorkshire, UK in September 2019. Photographs and a plan of the house are provided in Two different types of querns were used: a saddle quern and a rotary quern . These rWhile previous experiments on the effect of cooking or food processing tended to use only two to three grains/seeds , we opte3.2Environmental samples were taken to compare with the starch granule content of the mouthwash samples, recording the presence of both intact and altered starch granules. They enabled us to examine how starch granules travelled within the space, and whether granules from species ground earlier in the experiment remained in the environment.3.3All procedures outlined below were carried out in accordance with the Ethical Approval for Research on Human Tissues approved by the Biology Ethics Committee, University of York (reference ARA201907). Multiple samples were taken in order to examine the presence of intact and altered starch granules in the mouth and the environment. As the activity was conducted in a facility where tools may be in close proximity with the public, a careful cleaning protocol for the querns was adopted.Avena sp.), plant epidermal tissues and non-plant remains. The rotary quern control included mineral particles (including quartz), most of which probably derived from the quern stone and also examples of pollen and insect hairs. It was not possible to determine when these debris were deposited, and so this material could derive from the stones’ original/early use, or their storage conditions. The querns were then very carefully cleaned to avoid cross contamination. As the experiment aims to assess accidently inhaled airborne starch granules, it must be stressed that:1.Any residual material dislodged from the querns during processing would be insignificant in amount compared to the considerable quantity of fresh starch present as result of the experiment.2.Even if there is a evidence of a small amount of residual material being inhaled, this would still be evidence of inhalation during food processing.Control samples for microscopic analysis were taken from the querns during the first phase of cleaning, prior to experiments. This was to give an indication of any microremains, and in particular starch granules, that might have been present from their storage location. Samples taken both from the saddle quern and rotary quern prior to the start of the grinding experiment included plant remains, such as starch granules  to prevent debris alteration or degradation by preventing the activation of amylase in saliva . Prior tSlides for microscopic analysis were prepared in a clean laboratory where dust and dirt are monitored to reduce the risk of contamination to secure the integrity of the analysis. A 200 μl aliquot of the mouthwash containing the pellet was pipetted and mounted with 50:50 distilled water: glycerol onto a clean microscope slide, which was then immediately covered with a glass slide cover and sealed with clear nail varnish.Slides were analysed using an Olympus Microscope BX53M, under 200x magnification to gain an overview of the amount of debris. Slides were then examined under 400x magnification with all starch granules recorded and counted in a spreadsheet with the necessary photographs taken. Starch granules were classified as ‘intact’, ‘altered’, ‘other species’ and ‘nondiagnostic’. When needed, the magnification was increased to 630x. Observations were completed by scanning the samples with cross -polarised light, which allowed for birefringent material to be made visible and observe any alteration of the extinction cross that could be linked to damage by processing. Counting was not carried out for the samples of the ground grain taken from the querns due to time constraints. Any data gained from that analysis would not have added to the interpretations of this study. Samples were disposed of safely after analyses, following the standard method applied by the Archaeology Department for the disposal of biological material and following ethical guidelines.3.5The amount of time spent on each experiment by each individual is provided in Avena sativa) were counted per diagnostic compound starch granule (unless otherwise stated) as the compound nature of oat is diagnostic, unlike with other starch granules from the other grains which were counted per starch granule due to their non-compound nature.Below the control samples in 4Starch granules and microdebris identified from environmental and mouthwash samples are described by category below.4.1Starch granules in the flour produced during grinding were found to be present as a mixture of intact and damaged granules , the respiratory tract therefore may act as a reservoir for starch granules inhaled, increasing the chances of them to remain in the mouth for longer.3.A combination of both 1 and 2.The first control samples taken before the first grinding session (wheat on the rotary quern) had no starch granules present. The control samples taken prior to each grinding session thereafter had zero to six individual starch granules present in them, and these were never from the species that were to be ground during that session. The starch granules that were present in all other control samples were consistently possible wheat starch granules (i.e. A type and B type) and altered starch granules. Therefore, despite strict cleaning of teeth and mouth, some starch granules remained in the mouth, but in very low numbers. Three possible hypotheses are plausible:The results from the samples taken after grinding indicate that starch granules, both intact and altered, are inhaled via oral breathing during the grinding process. Only one mouthwash sample taken by one individual after grinding did not contain both intact and altered starch granules .The number of starch granules inhaled varied widely between the different querns, cereals and Individuals. Samples taken by Individual 2 consistently had a higher number of total starch granules present in each mouthwash sample compared to Individual 1. The most dramatic example for this is the case of millet ground on the saddle quern where the total number of millet granules identified for Individual 1 was three, whereas for Individual 2, the total was 248. For Individual 1, five out of six mouthwash samples had a higher number of altered starch granules identified than those from the species that was being ground. For Individual 2, however, intact starch granules from the species being ground were consistently found to be present at a higher rate than altered starch granules. The pie charts reflect this higher number of starch granules from ground species, especially as Individual 2 had consistently a higher number of identified starch granules in their mouthwash samples.Oat is present in fewer numbers compared to both wheat and millet, potentially indicating that the compound starch granules are less airborne than the single granules, and/or the small granules from broken oat compounds may be more subjected to salivary clearance. It is possible that as oat was counted in its compound form, this reduced the number of starch granules counted. However, these mouthwash samples also contained a generally low number of starch granules that were altered, derived from other species or nondiagnostic.55.11.Uncooked starch granules enter the human mouth during the grinding of all the grain types using quern stones. Some of this starch was also dispersed in the environment.2.The quantity of debris generated was not large, and the number of starch granules that entered the mouth were limited for each experiment, but they could accumulate if no mouthwash was undertaken.3.Many of the starch granules retrieved in the experiment were found to be intact.4.The number of starch granules that entered the mouth varied between species.5.Differences in the number of starch granules found in the salivary samples of the two researchers were different.Our experiment resulted in five key findings:5.2It has long been known that processing of large quantities of grains produces dust that can impact human health e.g. . Our expWhile the counts for diagnostic starch granules might appear to be quite low, certain factors need to be considered. Firstly, starch granules do not tend to be found in high counts in dental calculus samples compared to both the number present in plants themselves and the amount of plant foods an individual consumes in their lifetime . It shouA greater number of intact starch granules were generally found across both the mouthwash and environmental samples. This finding could indicate that altered starch granules are not as easily dispersed as intact granules, and intact granules tend to be dispersed quickly before more extended processing leads to them being cracked or broken. The results are unclear as to whether using a saddle quern (more exposed grinding) or a rotary quern  has an impact on the nature of the debris that is inhaled.Our results also indicate that there is much variation between individuals in terms of what and how much is inhaled via oral breathing, which would have an impact on how much debris has the potential to be trapped in dental calculus. This is due to a number of factors including whether someone is breathing through their mouth , how much they are talking, how often someone breathes out through their mouth , as well as salivary clearance rate.Wellcome Trust . Finally, The presence of starch granules in the environmental dust traps shows that a portion of the starch granules freed from the grains during grinding can reach the ground during processing. In many studies on stone tools, soil samples are often investigated for the presence of starch granules that can cause contamination; in such studies that presence of starch in soil is considered a potential agent of contamination of modern origin. This work suggests that some of such starch could actually be generated by the use of the stone tools and grinding implements, and therefore may not be strictly indicative of a contamination.There are also other non-human factors that may impact on debris being inhaled, such as the physical characteristics of the grain and processing time. As to be expected, starch granules from grain that was in the process of being ground were more likely to be inhaled. However, the results from the environmental and mouthwash samples show that starch granules from wheat are more likely to remain in the environment and continue to be inhaled after grinding. This might be due to wheat taking much longer to be ground (between 20 and 45 min for both querns), thereby extending the time period for these starches to be released in the environment. This experiment would need to be repeated and extended to examine these factors further. Such work is currently in progress for non-dietary remains funded by the 5.3The results of the experiment show that deliberate consumption is not the only pathway for intact and altered starch granules to enter the mouth and become entrapped in dental calculus. This is especially notable for intact starch granules, considering most of the current published experiments show that starch granules become altered when processed for consumption. Additionally, the ground grains (flour) sampled from the quern also showed that starch can remain intact after processing, further indicating that starch granules can survive grinding. This, in combination with mouthwash samples and environmental samples, shows that unaltered starch granules may be released into the environment, and while some may be quickly inhaled, much may remain.Our findings also show that the amount of starch granules retained in the mouth during grinding is very limited compared to the quantity of starch that would flood the oral cavity during eating. Additionally, it is possible that intact starch granules have a better chance to survive in the saliva and in the dental calculus matrix when compared to damaged starch, despite bacterial activity in the human mouth. Starch granule accumulation in the environment as a result of plant processing has been noted by 5.4It is important to consider whether different populations from different contexts would show variation. This study fits into broader research that is focused on the medieval period, comparing debris from dental calculus samples from a range of different sites across Medieval England, particularly between rural and urban contexts. In samples from both types of context in the overarching project, both intact and altered starch granules from cereals were found  in both rural and urban areas, would have experienced more intense prolonged exposure and presumably will have trapped a greater quantity of such debris in their dental calculus. Large scale production of flour in the past and today in many societies are known to have had a serious effect on human and animal respiratory health , and fur5.5Even though these experiments were carried out on a different typology of raw material, as well as different quantity and time of grinding in comparison to the lapis lazuli study by 5.6Finally, we would like to mention that the handling/use of human tissues, of which saliva is considered one, for research purposes are subject to strict ethical agreements, mainly aimed to protect any health information of the participants and the handling of DNA. However, wider ethical implications may be encountered if experimental work is conducted among groups where ethics agreements may not be as straightforward as it is in the case of this paper. We believe that experimental work conducted in this paper and by 6Our experiment has shown that the presence of intact starch granules in dental calculus can derive from grain processing rather than direct consumption alone. Furthermore, the results indicate that it is possible for both intact/diagnostic and altered starch granules to enter the mouth via oral breathing, during processing and even several hours after being ground.Due to the Covid-19 pandemic, more research is being conducted on the inhalation of particles from the surrounding environment. These studies show how particles can preserve, travel and survive in environments with regards to health, (for example: Experiments, like the one reported in this article, are necessary to improve understanding of past lifeways and to make interpretations of dental calculus microremains more accurate. We recommend that researchers look beyond the dietary consumption pathway alone when considering the origin of both intact and altered starch granules as inhalation can occur both through processing or even as a result of being present in an environment where cereal grinding has taken place, and other similar occupational pathways should also be considered when examining other types of debris.All authors contributed to the research design, with experimental protocols based upon on-going research by AR. SD and AR conducted the experimental work and laboratory analysis of oral samples. SD conducted the sampling and analysis of the environmental samples. SD with AR and MA conducted final data analysis and data contextualisation. SD, AR and MA wrote the paper.All data from this study are available in this article.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Due to its unique properties, zirconia is increasingly being used in dentistry, but surface preparation for bonding is difficult because of its polycrystalline structure. This study aimed to determine the effect of a new etching technique (Zircos-E) on Ceramill Zi (Amann Girrbach). The effect of etching and the use of primers (Monobond Plus and MKZ Primer) on the bond strength of zirconia with resin cement (NX3) was assessed. Shear bond strength was evaluated after storage in water for 24 h and after thermal aging (5000 thermocycling at 5 °C/55 °C). A scanning electron microscope (Hitachi S-4700) was used to evaluate the surface structure before and after the Zircos-E system. The roughness parameters were assessed using an SJ-410 profilometer. The etched zirconia surface is more homogeneous over the entire surface, but some localized forms of erosion exist. The etching of zirconia ceramics caused changes in the surface structure of zirconia and a significant increase in the shear bond strength between zirconia and resin cement. The use of primers positively affects the adhesion between resin cement and zirconia. Aging with thermocycler significantly reduced the shear bond strength, with one exception—sandblasted samples with MKZ Primer. Standard ceramic surface preparation, involving only alumina sandblasting, does not provide a satisfactory bond. The use of etching with the Zircos-E system and primers had a positive effect on the strength of the zirconium–resin cement connection. Yttrium-stabilized zirconia (Y-TZP) is most often used in dentistry due to its combination of high strength and optimum optical properties [0.5) in comparison with other conventional ceramics, including alumina-reinforced and lithium-disilicate-based ceramics [The most popular ceramics used in dental practice are lithium disilicate, silica, alumina, leucite, and zirconia-based materials. However, zirconia is favored both in research and in the clinic. The number of research publications related to this type of ceramic nearly tripled in the years 2012–2019 compared to 2007–2011 [ceramics . Becauseceramics .Success in prosthetic treatment is not only related to selecting the appropriate material for the reconstruction, but requires the creation of appropriate adhesion. Unfortunately, due to the crystalline structure of zirconia and the lack of a silicon dioxide (silica) phase, conventional methods of preparing dental ceramics (including etching with acidic solutions and applying silane coupling agents) do not result in sufficient bond strength in restorations . Differe2SO4), nitric acid (HNO3), and phosphoric acid (H3PO4). This patented technology uses ionization to create a microporous surface, which may improve bonding strength [Nevertheless, the surface of zirconia should be prepared before bonding with any cement in order to promote micromechanical interlocking mechanisms. The durability and stability of the bonds between ceramics and adhesives can be improved because the physical bonds are not susceptible to the degradation caused by environmental factors, such as humidity . Some austrength . In the strength . The first null hypothesis was that etching with the Zircos-E system or using primers (MKZ Primer or Monobond Plus) does not affect bond strength. The second was that the surface topography of Ceramill Zi  is not affected by etching with the Zircos-E system .2O3 particles under a pressure of 2 bar. After that, the specimens were washed under running water for 30 s and then air-dried. Half of the specimens were etched for 30 min at 30 °C using Zircos-E Etching Solution . Zircos-E Etching Solution contains hydrofluoric acid (HF), hydrochloric acid (HCl), sulfuric acid (H2SO4), nitric acid (HNO3) and phosphoric acid (H3PO4). After the etching process, the samples were cleaned with running water for 3 min and then with steam.The monolithic zirconia CAD-CAM blocks  were sectioned using a milling device . Subsequently, the specimens were sintered according to the manufacturer’s instructions. The samples were cylinder-shaped with a diameter of 10 mm and a height of 10 mm. The surface was ground under water coolant with 180 grit, followed by 320 and 600 grit silicon carbide paper, to unify sample surfaces. Prepared specimens were sandblasted with 50 μm Aln = 22), as follows: Control: No chemical treatment was conducted.Primer MKZ: MKZ Primer  was applied according to the manufacturer’s instructions.Primer Monobond Plus: Monobond Plus  was applied according to the manufacturer’s instructions.Etched: No additional treatment after etching was conducted.Etched + Primer MKZ: After etching, MKZ Primer was applied according to the manufacturer’s instructions.Etched + Primer Monobond Plus: After etching, Monobond Plus was applied according to the manufacturer’s instructions.According to the conducted surface preparation, the specimens were divided into 6 groups .OptiBond Solo Plus (bonding agent) was applied according to the manufacturer’s instructions on prepared samples. Next, composite cement  was packed into a plastic mold with a diameter of 3 mm and a height of 3 mm and light-cured for 20 s using a light-curing unit . After that, the shear bond strength was evaluated.The roughness of the control and etched samples was investigated using a compact surface roughness tester SJ-410 . Surface roughness for each study group was measured five times in different directions, from which the mean values were calculated. The evaluation length was 4 mm. Six roughness profile parameters were compared: roughness average (Ra), the average maximum height of the profile (Rz), maximum profile peak height (Rp), maximum profile valley depth (Rv), mean spacing of profile irregularities (Sm), retention volume (Vo).Control and etched materials’ surface topography was investigated with a scanning electron microscope energy-dispersive spectroscopy (SEM–EDS) microscope Hitachi S-4700 . The images were made at a magnification of 1000×, 5000× and 15,000×.t-test to compare parameters  of control and etched samples. All analyses were performed using Statistica version 13 software . The value α = 0.05 was adopted as significant. Shear bond strength data were analyzed by the Shapiro–Wilk test to assess the normality of the distributions. Based on the results, the Kruskal–Wallis test with multiple comparisons of mean ranks was applied. Data obtained from roughness measurements were subjected to Student’s The results of shear bond strength after 24 h in water and thermocycling are presented in p-value ≤ 0.0000. The highest median value of shear bond strength evaluated after 24 h in water was 17.7 MPa (etched+ Primer Monobond Plus), and the lowest was 6.1 MPa (control) . The usep = 0.000434);Control vs. Etched (p = 0.000001);Control vs. Etched + primer MKZ (p = 0.000001);Control vs. Etched + primer Monobond Plus (p = 0.003873);Primer MKZ vs. (b) Etched + primer MKZ ((p = 0.004601).Primer MKZ vs. Etched + primer Monobond Plus According to multiple comparisons of mean ranks for 24 h, water series, statistically significant differences were found between:p = 0.000114);Control vs. (x) primer MKZ (p = 0.000069);Control vs. Etched (p = 0.000059);Control vs. Etched + primer MKZ (p = 0.00075).Control vs. Etched + primer Monobond Plus ; parameter (p < 0.05).According to Student’s The surface topography of control and etched samples at a magnification of 1000×, 5000×, and 15,000× is presented in The images obtained with the SEM evaluation show clear differences between the surface topography of the two materials. Both samples show an isotropic structure; however, the etched sample is more homogeneous, with regularly and evenly distributed crystal grains. Ceramill Zi is composed of zirconium oxide and the signals corresponding to zirconium (Zr) and oxygen (O) are mainly visible in the analyzed spectra A,B. The Manufacturing a zirconia prosthetic restoration poses a challenge, as it requires the creation of an appropriate connection with resin cement. Traditional etching methods are ineffective because the material is polycrystalline and has no silica phase compared to conventional ceramic materials. Our findings, based on analyses of shear bond strength, suggest that the etching (Zircos-E system) and presence of primers (MKZ Primer or Monobond Plus) influence the adhesion of resin cement (NX3). SEM micrographs showed differences in the structure after etching. Therefore, the null hypothesis can be rejected. All samples were first sandblasted (control sample). This treatment caused isotropic roughening and irregularities formed on the surface . The str2O3; etched with Zircos-E; air-abraded and then etched with Zircos-E; etched with Zircos-E and then sandblasting) were subjected to thermocycling for 1000 cycles between 5 °C and 55 °C. Similar results (around 10 MPa) were obtained regardless of the surface preparation method. The authors suggested that an additional study should be performed, nothing the potentially hazardous nature of such an acid solution [Treatment with the Zircos-E system improved the bond strength compared to the sandblasted samples. Observed changes in surface topography and smaller distances between irregularities on the surface result in the better micromechanical interlocking of resin cement. A similar dependency can be found in other studies . In the solution . In anotsolution . The autsolution .In our study, the application of selected primers also increased bond strength. This increase was, however, statistically insignificant. MKZ primer contains 3-Methacryloyloxypropyl-trimethoxysilan and 10-Methacryloyloxydecyl dihydrogen phosphate. Monobond Plus has three different functional methacrylates—silane, phosphoric, and sulfide methacrylate. Silanes can react with hydroxyl groups of silica and the methacrylate of resin cement, creating quite a strong connection with inorganic ceramics. However, in zirconia, there is no silica phase, so this bonding cannot occur with silane substances of used primers ,39. HoweLong-term water storage and thermocycling are common methods of assessing bond durability ,45,46. IIn summary, our study confirms that zirconia can be etched. It also shows an increase in shear bond strength. The observed changes in SEM images may be related to the corrosion of zirconia grains. More chemically reactive atoms around the crystal boundaries dissolve faster than those inside the crystal, which leads to the formation of irregular grooves around the crystals and grain size reductions . It was Further studies on different types of cement and variations in the etching protocol  would provide a more detailed insight into the effect of the Zircos-E Etching solution on zirconia to improve bond strength. A comparison of the use of silane alone would allow for an evaluation of the effectiveness of this substance in comparison with primers dedicated to zirconium (containing other monomers). Testing mechanical properties, such as hardness and flexural strength after such aggressive etching, should be evaluated in the future for zirconia ceramic. In addition, an evaluation of the effect of certain treatments on zirconia phases with the use of X-ray diffraction and FITR would be an interesting contribution to research on zirconia.Use of the Zircos-E system influences the shear bond strength between zirconia and resin cement. In consequence, SBS is significantly higher than that observed in the control group (only sandblasted zirconia) and when only a primer (after sandblasting) is used.Etching with the Zircos-E system leads to changes in the surface structure—a lower average roughness and mean spacing of profile irregularities. Etching also reduces the amount of alumina element (from air abrasion) on the zirconia surface.Use of primers preceded with etching with the Zircos-E system positively affects the adhesion between resin cement and zirconia.Aging with thermocycler significantly reduced the shear bond strength, except for sandblasted samples with MKZ Primer, which stayed on the same level ~10 MPa, before and after aging.Considering the limitations of the present study, the following conclusions can be drawn:


Protein–starch interaction has an important impact on the properties of starchy foods rich in protein, but the contribution of the interaction to Chinese yam still remains unclear. This study aimed to characterize the physicochemical and functional properties related to the possible interaction between starch and protein in Chinese yam. Differential scanning calorimetry and rapid viscosity analyzer results revealed that the gelatinization temperature increased in protein and starch cross‐linked powder, while the peak viscosity and the setback viscosity decreased. The swelling power and solubility at 80°C and 95°C decreased with increasing protein ratio in the powder. In vitro starch digestibility test indicated that a high protein ratio could rapidly reduce digestible starch, but increase both slowly digestible starch and resistant starch. Protein could act as the physical barrier toward starch against heating and digestion to exert the influence on starch properties. Fourier transform infrared spectroscopy test revealed the interaction between protein and starch. These results revealed the role of protein–starch interaction and provided beneficial information for the utilization of Chinese yam. The study aimed to characterize the interaction between starch and protein and the changes in the physicochemical properties and functional properties of yam. Protein and starch content of the yam powder was 14.0% and 78.0%, respectively.Yam powder was prepared referring to Xie et al. . The yam2.3The method of our previous study , and the mixtures were evenly vibrated and shaken. For subsequent tests, the yam powder or mixtures were stirred in water for 10 min to obtain sufficient dispersion.2.6To), the peak temperature (Tp), the conclusion temperature (Tc), and the crystal melting enthalpy (ΔH).The gelatinization properties of samples were analyzed by Mettler DSC 1 professional thermal analyzer  according to Shao et al. . The sam2.7A modular compact rheometer  was used to measure the pasting properties according to Shao et al. . The pas2.8g for 15 min, the supernatant was dried at 105°C for 2 h  was heated in a shaking water bath at 80°C or 95°C for 30 min and then cooled to room temperature. After centrifugation at 1699 The swelling power and solubility of the powder were calculated as follows:2.9The method of Englyst et al.  was appl2.10About 500 mg of powders and 5 ml of distilled water were mixed, stirred, and heated in 100°C water bath for 15 min to obtain a paste. Powders were double‐stained and pasted with fluorescent dyes  in the dark for 5 min. The stained section was covered carefully with a cover glass. Then, fluorescence images were captured with confocal laser scanning microscopy . Two excitation wavelengths of 543 nm helium–neon laser and 488 nm argon–krypton laser were used to reflect the fluorescence of protein and starch, respectively . Samples were prepared using 6% (w/w) powder dispersed in KBr pellets. All powders were dried at 105°C for 24 h, then compressed with KBr, and 256 scans were recorded with a resolution of 4 cm2.12p value was <.05, the value was considered significantly different.Data were expressed as means ± standard deviation of three replications and performed one‐way analysis of variance (ANOVA) with Duncan's multiple range test (SSR) for statistical analysis by SPSS 25.0 statistical software . When the 33.1To), peak temperature (Tp), and conclusion temperature (Tc) of all powders had an increasing tendency. The mixture of 20% protein had the highest To, Tp, and Tc, which were 73.8, 76.8, and 80.8°C, respectively. However, accompanying the increase in protein content, the gelatinization enthalpy (ΔH) of the powder was significantly reduced. The powder with a protein ratio of 20% had the lowest gelatinization enthalpy of 10.0 J/g.Gelatinization properties with DSC are shown in Table Starch granules absorbed water and expanded under the action of moisture and heat, which would be accompanied by the leaching of small granules . Compared with pure starch, the RDS content of starch–protein samples decreased significantly, and starch digestion was slowed down in a short time. The increase in SDS content indicated that the suppression of the mitigation effect continued. As the protein content increased, the RS starch fraction steadily increased, which meant that the degree of starch digestion decreased. This may be related to the fact that more protein matrix wrapped the starch network, which limited the efficiency of starch decomposition. The reduction in digestion efficiency was reflected in the simultaneous decline in speed and degree. The protein wrapped starch granules and may become an obstacle to delay enzymatic hydrolysis  images of fresh and cooked yam powder with different protein–starch ratios are shown in Figure As shown in Figure As shown in Figure 3.6−1 were related to the short‐range structure and hydrated crystals of starch  and amide II (1480–1580 cm−1) (Li et al., The FTIR spectrum could show the microstructure of the substance through characteristic peaks. The analysis of yam powders is shown in Figure −1 of powders was evaluated in Figure −1 was related to the ordered and amorphous structure of starch, and the other band at 995 cm−1 was related to hydrated crystals (Sevenou et al., In order to further study the influence of protein on the short‐range structure of starch, the ratio of 1047/1022 cm4There was a hydrogen bond between protein and starch, mainly due to their interaction, which had a great impact on the physicochemical and digestive properties of the yam starch. Increased protein in the starch could block the process of starch gelatinization and retrogradation with reduced peak viscosity and final viscosity. The swelling power of starch at 80°C and 95°C was decreased, and the water absorption performance was negatively affected by protein. The addition of protein reduced the RDS but increased the SDS and RS. Protein–starch interactions resulted in the formation of a granule mixture, which acted as a physical barrier to starch. With the addition of protein, the short‐range ordered structure of yam powder was significantly enhanced. The physicochemical and functional properties of yam starch were clearly influenced by existing protein.The authors declare no competing interests.


Thermoplastic starch was blended with commercially available biodegradable polyesters of poly (PBAT) and poly(lactic acid) (PLA) for its improved performance and processability. The morphology and elemental composition of these biodegradable polymer blends were observed by scanning electron microscopy and energy dispersive X-ray spectroscopy, respectively, while their thermal properties were analyzed using thermogravimetric analysis and differential thermal calorimetry. For rheological analysis, the steady shear and dynamic oscillation tests of three samples at various temperatures were investigated using a rotational rheometer. All three samples exhibited significant shear thinning at all measured temperatures, and their shear viscosity behavior was plotted using the Carreau model. The frequency sweep tests showed that the thermoplastic starch sample exhibited a solid state at all temperatures tested, whereas both starch/PBAT and starch/PBAT/PLA blend samples exhibited viscoelastic liquid behavior after the melting temperature such that their loss modulus at low frequencies was greater than the storage modulus, and inversion occurred at high frequencies (storage modulus > loss modulus). With increasing attention given to global environmental issues, starch-based biodegradable polymer materials have attracted considerable attention as a substitute for substantial conventional petrochemical-based synthetic plastics. The development of biodegradable plastics has long been a major issue in the polymer community to reduce the impact of plastic waste on the environment. Starch is a natural biodegradable polymer that is abundant in plants such as corn, wheat, potato, and rice. In addition, starch is considered a promising candidate for developing biodegradable plastics because of its complete biodegradability, renewability, low cost, and low density ,2. On thAmong these, the PBAT is a biodegradable aliphatic–aromatic copolymer . Starch/Concurrently, PLA is a well-known biodegradable thermoplastic polyester that can be produced from agricultural products such as corn and sugar beets . It is c3) and carbon black [Moreover, various studies have been performed regarding their thermorheological properties. Using thermoplastic starch (TPS), Gonzalez et al.  showed ton black ,23. Moreon black ,25. Furt®E100  and hydroxypropyl cellulose, the increased temperature reduces the shear viscosity of each polymer and its blends, implementing the process of pharmaceutical industry such as injection molding [In many polymer processing operations, such as extrusion, blow, and injection, the rheological properties are important for controlling processing conditions and the quality of the final product . The mel molding . Moreove molding .This study examined the steady shear and dynamic oscillatory rheological properties of starch-based biodegradable plastics, including starch and glycerin, starch and PBAT blend, and three-component blend with starch, PLA, and PBAT. Their surface morphologies were observed using scanning electron microscopy (SEM), and the chemical element composition was confirmed by energy dispersive X-ray spectroscopy (EDS). The thermal performance was analyzed using thermogravimetric analysis (TGA) and differential scanning calorimetry (DSC). Their melt rheological properties under various temperature conditions were studied using a rotational rheometer. Biodegradable ternary systems in this study could possess potential commercial applications in various areas, including agricultural mulching films, disposable plastic bags, disposable trays, disposable straws, and disposable plastic tableware. In addition, rheological data of the ternary blend system in this study would be beneficial for their compounding.Three commercial starch-based bioplastic materials called Tapioplast (S1), BioBlend AG202050 (S2), and Rodenburg compound (S3) were purchased from SMS Corporation , BioLogiQ , and Rodenburg Biopolymers , respectively. Based on the information from each of the manufacturing companies, it was found that the composition of S1 is known to be more than 75% of tapioca starch and less than 25% of glycerol. S2 and S3 are blends of starch and biodegradable polymers. S2 contains mainly starch and PBAT with about 30% content of PBAT. S3 mainly contains starch, PBAT, and PLA at 16%, 18%, and 45%, respectively. In addition, some unknown additives might be added to improve their compatibilization or/and mechanical properties.The morphologies of the three samples were observed by SEM , and their element compositions were examined using SEM-equipped EDS. The thermal characterizations, including thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC), were performed using a thermal analysis system . Their melt rheology was first tested by producing disc-shaped samples with a thickness of 1 mm and a diameter of 25 mm using a hot-pressing machine. The rheological properties at various temperatures were tested using a rotational rheometer  with a parallel plate geometry (PP 25).2 atmosphere. The weight loss of S1, S2, and S3 occurs through two, three, and four steps, respectively. The weight lost before 190 ℃ was attributed to the evaporation of water. As shown in 2 atmosphere. The glass transition temperature (Tg) of S1 was approximately 78 °C, and no melting peak was observed. S2 showed a melting peak at 118 °C, which was attributed to the presence of PBAT, because PBAT is composed of two parts, butylene terephthalate (BT) and butylene adipate (BA), and their melting temperatures (Tm) were approximately 60 °C and 130 °C, respectively [The DSC curves  of S1, Sectively . Therefoectively .The melt rheological properties of the samples were examined by first performing a temperature sweep test at a heating rate of 4 °C/min with a strain of 1% and an angular frequency of 6.28 rad/s. After checking melting temperature (Tm) by the temperature sweep test, the strain amplitude test and steady shear test were performed under various temperature conditions around the Tm temperature. The shear stress and shear viscosity as a function of the shear rate under various temperatures were tested in controlled shear rate (CSR) mode. As shown in n < 1 and n = 1, it indicates Newtonian flow. The solid lines in In addition, various models have been developed to measure shear viscosity at high shear rates owing to the difficulty in practical measurements . The CarThe dependence of the material on frequency was examined by performing the frequency sweep test at a fixed strain of 0.01% for each temperature. As shown in G′ and G″ are not constants but change with time because viscoelastic materials are time-dependent. In the dynamic oscillation tests, the time dependence can be evaluated by changing the frequency of the applied stress or strain. Three starch-based biodegradable materials of starch, starch/PBAT blend, and starch/PBAT/PLA blend were characterized using SEM, EDS, TGA, and DSC. The melt rheological properties, including steady shear and dynamic oscillation tests at various temperatures, were investigated using a rotational rheometer. SEM revealed the two blend systems of S2 and S3 to have good phase compatibility. Using EDS and TGA, it was speculated that there were calcium-containing additives in S3, and the content of starch in S3 was relatively small. S1 showed a wide glass transition region, and no melting peak was observed. Affected by the melting of PBAT, the glass transition process of S2 was divided into two stages, and a melting peak appeared at 118 °C. Owing to the presence of PLA, the glass transition process of S3 appeared in two stages, and there were three melting peaks. In the melt rheological analysis, S1 failed to reach the molten state until 220 °C, and the melting temperatures of S2 and S3 were 159 °C and 150 °C, respectively. The storage modulus, loss modulus, shear stress, and shear viscosity decreased with increasing temperature. All samples exhibited significant shear thinning at all temperatures, and the shear viscosity behavior of all products at a high shear rate can be predicted by the Carreau model. The frequency sweep tests showed that S1 exhibited a solid state at all tested temperatures, while S2 and S3 exhibited viscoelastic liquid behavior after the melting temperature. In addition, in the case of S2 and S3, the loss modulus at low frequencies was greater than the storage modulus, and inversion occurred at high frequencies (storage modulus > loss modulus). Furthermore, the frequency of intersection became higher at higher temperatures, which means that the relaxation time became shorter. Therefore, considering these thermorheological properties within high range of temperature, S1, which showed solid-like properties at high temperature, can be applied in high-temperature thermo-processing, while due to its low Tm, it is easier to carry out the molding process at a relatively low temperature during processing.


T+Itpr3tf/J (BTBR/J) strain is one of the most valid models of idiopathic autism, serving as a potent forward genetics tool to dissect the complexity of autism. We found that a sister strain with an intact corpus callosum, BTBR TF/ArtRbrc (BTBR/R), showed more prominent autism core symptoms but moderate ultrasonic communication/normal hippocampus-dependent memory, which may mimic autism in the high functioning spectrum. Intriguingly, disturbed epigenetic silencing mechanism leads to hyperactive endogenous retrovirus (ERV), a mobile genetic element of ancient retroviral infection, which increases de novo copy number variation (CNV) formation in the two BTBR strains. This feature makes the BTBR strain a still evolving multiple-loci model toward higher ASD susceptibility. Furthermore, active ERV, analogous to virus infection, evades the integrated stress response (ISR) of host defense and hijacks the transcriptional machinery during embryonic development in the BTBR strains. These results suggest dual roles of ERV in the pathogenesis of ASD, driving host genome evolution at a long-term scale and managing cellular pathways in response to viral infection, which has immediate effects on embryonic development. The wild-type Draxin expression in BTBR/R also makes this substrain a more precise model to investigate the core etiology of autism without the interference of impaired forebrain bundles as in BTBR/J.The BTBR  Autism spectrum disorder (ASD) is a neurodevelopmental disorder with complex genetic architecture and heterogeneity, which have impeded the development of therapeutic strategies for this disease with a 2.3% prevalence rate in 2018 , 5. ThesT+Itpr3tf/J (hereafter referred to as BTBR/J) has been recognized as an idiopathic model which well recapitulates the core symptoms of autism, including impairments in social interaction, repetitive behaviors, and often associated ultrasonic communication deficits [From a phenotyping screen among several inbred strains, BTBR deficits –8. It isdeficits , 8, showdeficits . Togethedeficits , 4. BTBRdeficits , 11. HowT+Itpr3tf/J (BTBR/J) , were purchased from Japan SLC Inc. , RIKEN BioResource Research Center , and the Jackson Laboratory, respectively. All three strains were maintained in the same breeding room under controlled temperature at 23 ± 0.5 °C and humidity of 50–60% with 12 h light-dark cycle (light on 8 am) and ad libitum access to water and food. All procedures for animal handling followed the Animal Experimentation Committee of RIKEN Brain Science Institute guidelines.C57BL/6J and two BTBR strains, BTBRTF/ArtRbrc (BTBR/R)  and BTBR aCGH was performed according to the manufacture’s protocol . Briefly, genomic DNA (gDNA) was extracted from the tails of male mice by using Blood & Cell Culture DNA Mini Kit (Qiagen). The quality of DNA was checked for the absorbance at 320 nm, the ratio of OD260/280 > 1.8 and OD260/230 > 1.0. The gDNA was digested and labeled with Cy3- or Cy5-dCTP by random priming . After hybridization, the fluorescence signals were scanned and analyzed by using DNAcopy, Gviz, and GenomicRanges packages under the R environment .Nine-week-old male mice of both BTBR strains and B6 were used for the MRI study. The mice were anesthetized for transcardiac perfusion at the speed of 1 ml solution per minute. They were first flushed with 30 ml of PBS containing 1 μl/ml heparin, 2 mM ProHance, and then fixed by 30 ml of 4% paraformaldehyde in PBS with 2 mM ProHance at room temperature. The brain remained in the skull with the tissue, zygomatic bone, and the lower jaw removed. After dissection, the remaining skull structure was placed in a 15 ml tube filled with fixation solution overnight at 4 °C. On the next day, each specimen was preserved in PBS with 0.02% sodium azide and 2 mM ProHance and kept at 4 °C for at least one month until MRI scanning . A multi3 and matrix size equaled to 504 × 504 × 630. Voxel dimensions for this scan were 0.040 mm isotropic. The total imaging time was ~14 h.In order to detect volumetric changes, the following parameters were used for the MRI scan, which was a T2- weighted, 3-D fast spin-echo sequence with a cylindrical acquisition of k-space, TR of 350 ms, TEs of 12 ms per echo for six echoes, field-of-view equaled to 20 × 20 × 25 mm3 and a matrix size of 180 × 180 × 324 yielding an image with 78 μm isotropic voxels. Five b = 0 s/mm2 images and 30 high b-value (b = 2147 s/mm2) images in 30 different directions were acquired using the Jones30 scheme [Diffusion Tensor Imaging (DTI) was done using a 3D diffusion weighted fast spin echo sequence, with an echo train length of 6. Parameters for the DTI sequences were TR = 270 ms, first TE = 32 ms, and a TE of 10 ms for the remaining five echoes, 1 average. Field-of-view of 14 × 14 × 25 mm0 scheme . Total ib = 0 s/mm2 images from DTI) are linearly (6 followed by 12 parameters) and non-linearly registered together. Registrations were performed with a combination of mni_autoreg tools [b = 0 s/mm2 DTI images are registered separately. The result of this registration is to have all images deformed into alignment with each other in an unbiased fashion.To visualize and compare any changes in the mouse brains, the anatomical images  for RNA extraction following the manufacturer’s instructions. RNA was treated with DNase I (Promega) and purified again by phenol/chloroform. 2.5 μg of total RNA was used for reverse transcription with random primers by SuperScript® II (Invitrogen). The PCR reaction was performed with 2 μl of 1/10 diluted cDNA template, specific primer pairs, and Power SYBR Green Master Mix (Thermo Fisher) in StepOnePlus TM (Thermo Fisher). Relative expression was calculated after normalizing to the endogenous control gene, Gapdh  were analyzed for their USV calls during a 5-minute separation from their mothers. Each pup was placed onto fluffy bottom bedding in a glass beaker and moved into a dark, soundproof box. The USV calls were recorded using an ultrasonic microphone connected to a pre-amplifier set at a 250 kHz sampling rate . The acoustic data were digitalized by an Avisoft signal conditioner and recorded with Avisoft-Recorder software during the recording. The call number and types of the first 3 min of recording were analyzed manually according to the previous study .The subject was first put into a clean and transparent cage without bedding for 10-min habituation and followed by a 10-min recording from the side view. The light intensity in the cage was around 30 lux. The total grooming time was analyzed manually.A clean cage with 5 cm ALPHA-dri bedding  was prepared, and 20 clear blue marbles were set on the bedding. Each subject was put into the cage for 30 min. At the end of the test, the subject was removed from the cage. The location of the marbles was photographed and classified into still (at the same position), moved (move away but not buried), half (~50% covered with bedding), and buried (>50% covered with bedding). The light intensity at the cage was ~30 lux.The test arena is a rectangular three-chambered box divided by two Plexiglas walls with small square openings (5 × 3 cm), which allow the subject to enter each chamber . Each chamber was 20 × 40 × 22 cm in size, and each of the two side chambers had a small wire cage in quadrant shape at the corner. The arena was illuminated at 20 lux. The subject was first put into the central chamber and allowed to explore the entire 3-chamber box for habituation freely. Followed by a 10-minute habituation, in the first testing section, an age-matched unfamiliar mouse (S1) of the same strain or an inanimate object of comparable size was put into the wire cages on either side. The position of the stranger mouse and object changed alternatively for each subject. In the second section of preference for social novelty, the inanimate subject was replaced with another unfamiliar mouse (S2). The subject mouse was allowed to explore the familiar stranger (S1) or novel stranger (S2). Each section was 10 min, and the movement of each subject was recorded from the top view to analyze the time spent in around the wire cage, which was used to indicate whether the subject preferred a social object  or a novel social object .https://ohara-time.co.jp/products/barnes-circular-maze/).The circular open field was 1 m in diameter with 12 holes equally spaced around the perimeter  and elevated 75 cm above the floor. A black Plexiglas escape box (17 × 13 × 7 cm), covered by a layer of clean bedding, was set below one of the holes. The position of the escape box and target site was fixed and randomized for each mouse. Four visual cues were hung around the maze. The subject was released from the center to explore the maze for three trials per day and 6 successive days. The movement of the subject was recorded and analyzed by video tracking software  gene mutation, which is still carried by the BTBR/R strain but dropped in BTBR/J  and Itpr3 (tufted) genes, respectively. BTBR/R mice also show faster growth and larger body weight/size than B6 from the postnatal stage, as previously observed in BTBR/J [The appearance of the two BTBR strains was indistinguishable, except for the short-tailed phenotype due to a n BTBR/J .T mutation (T t/T t) is known to be embryonic lethal since the T gene encodes a T-box transcription factor crucial for the developmental process [T +/T t) from the comparative analysis to avoid any unknown dosage effect of T mutation, which could complicate the comparison to BTBR/J (T +/T +).Unexpectedly, we found that BTBR/R has an intact corpus callosum in both shortened- and normal-tailed mice  transposon, also known as endogenous retrovirus (ERV), was enriched threefold (30%) Fig. , top in To further investigate the involvement of ERV in the accelerated CNV accumulation in the BTBR strain, we next compared the repeat sequence in the shared CNVs with different copy numbers within the two strains, which suggest potential hotspots for CNV formation. To identify the shared CNVs with different copy numbers, a second aCGH analysis was performed by comparing the genome of BTBR/R or BTBR/J to the B6 strain as a reference  suggests double-stranded RNA virus infection activates the integrated stress response (ISR), disrupting protein synthesis in the fetal brain . In addiTo fight against the stress from virus infection, the host cells have evolved defensive mechanisms, such as ISR, which leads to global attenuation of cap-dependent translation to slow down virus invasion. On the other hand, some viruses have counteracting strategies against ISR  and hijaRpl29 expression in testis , suggesting recurrent CNVs generated by NAHR events. The distributions of highly homologous pairs of ERV elements are known to determine the susceptibility regions for recombination events . One of For a successful infection, viruses must employ the host cell’s protein synthesis machinery to restrict immune response and produce viral components for their survival and spread. On the other hand, the host also has evolved defensive mechanisms, such as ISR, to downregulate RNA translation to slow down the viral invasion. This phenomenon is observed in the Poly(I:C)-induced MIA model, suggesting the synthetic double-stranded RNA might directly activate the ISR pathways in the cells of fetal brains . On the In addition to being the building blocks for human (8%) and mouse (10%) genomes, the 5′ and 3′ long-terminal repeats (LTRs) of ERV are enriched with transcription factor-binding site, which serve as critical regulatory elements to control its own activity and the expression of nearby host genes , 83. It Escaping from the epigenetic silencing, active ERV in germ cells accelerates de novo CNV formation, which increases genome instability and makes BTBR strains a still evolving multiple-loci model of autism . MeanwhiSupplemental text and figuresSupplemental table 1Supplemental table 2Supplemental table 3Supplemental table 4Supplemental table 5Supplemental table 6


ML) regrets having to report the death of one of its most illustrious members, Dr. Josep Maria Queraltó, 67 years old, who left us suddenly, on Friday, March 27, at his home in Sant Andreu de Llavaneres (Barcelona).The Spanish Society for Laboratory Medicine .In addition, he was a member of the Commission on the Diagnostic Value of Biochemical magnitudes (1985–1989) and of the Scientific Committee of the Society (1985–1992), as well as of the Editorial Committee of the Journal Advances in Laboratory Medicine and the Board of Directors; he was still holding the last position at the time of his death.ML in international organizations has been of great importance, as he held highly significant positions, such as the presidency of the board of the European Federation of European Societies of Clinical Chemistry. He was also a member of the EC4 European Registry Committee from 2000 to the present.Likewise, his work representing SEQCML to be present at numerous international events. Among these it is worth noting, due to its great impact, the organization of the congresses EuroMedLab Barcelona 2003, of which he was president of the Scientific Committee, and EuroMedLab Barcelona 2019, of which he was a member of the Organizing Committee. The success of both European Congresses gave the SEQCML an enormous boost and visibility at an international level.His perseverance in the many international tasks in which he participated and his international contacts, which he cultivated and maintained exquisitely, helped the SEQCIn the days since his death, the Society has received dozens of emails and written statements from various countries around the world, expressing sorrow and dismay at the loss of such a beloved and valued person. Within his great human and professional worth, all those who had the opportunity to interact with him highlighted his patience and his great vision for the future in the field of Laboratory Medicine training.


Self-sensing actuation of shape memory alloy (SMA) means to sense both mechanical and thermal properties/variables through the measurement of any internally changing electrical property such as resistance/inductance/capacitance/phase/frequency of an actuating material under actuation. The main contribution of this paper is to obtain the stiffness from the measurement of electrical resistance of a shape memory coil during variable stiffness actuation thereby, simulating its self-sensing characteristics by developing a Support Vector Machine (SVM) regression and nonlinear regression model. Experimental evaluation of the stiffness of a passive biased shape memory coil (SMC) in antagonistic connection, for different electrical  and mechanical input conditions  is done in terms of change in electrical resistance through the measurement of the instantaneous value. The stiffness is then calculated from force and displacement, while by this scheme it is sensed from the electrical resistance. To fulfill the deficiency of a dedicated physical stiffness sensor, self-sensing stiffness by a Soft Sensor  is a boon for variable stiffness actuation. A simple and well-proven voltage division method is used for indirect stiffness sensing; wherein, voltages across the shape memory coil and series resistance provide the electrical resistance. The predicted stiffness of SVM matches well with the experimental stiffness and this is validated by evaluating the performances such as root mean squared error (RMSE), the goodness of fit and correlation coefficient. This self-sensing variable stiffness actuation (SSVSA) provides several advantages in applications of SMA: sensor-less systems, miniaturized systems, simplified control systems and possible stiffness feedback control. The shape memory coil (SMC) has a larger change in force and controllable stiffness to introduce the structural elastic deformation and to be in tune with a structural load. It provides the actuation to a mechanical structure; actuation with variable load can be sensed by self-sensing the stiffness in the structure via the Shape Memory Alloy (SMA) coil’s resistance. This is because of the shape memory effect phenomenon, which is an inherent property present in the nickel-titanium alloy. This inherent property is due to phase transformation from martensite to austenite and vice-versa when subjected to temperature or current. Though the SMA has same chemical composition, atomic weight and mass number, but it is different structure in the austenite and martensite phase.Until now, none is dedicated to physical sensor or analytical models to sense the stiffness of shape memory alloy or SMA with mechanical structure. Nowadays, the sensing function of SMA becomes more important because the high technology for the humanoid robot, industrial automation and medical field is being progressed a lot. It has been found 30 years ago that the electrical resistance of SMA changes during phase transformation of its material . The modThis electrical property is useful for sensing of thermal and mechanical properties like temperature, force and strain of SMA. Since then, many research papers have proved that resistance change is sufficiently linear to measure and control displacement and strain . These wThe polynomial model accurately enables estimation of the SMA actuator strain by applying an electrical potential across it. The experimental results have shown that the self-sensing model can achieves a small transient error and works effectively. The self-sensing helps to develop to miniaturized devices to perform effectively and efficiently  in whichFrom the results of this work, it has been realized that the use of neural networks to characterize the relation between the resistance of SMA wire and its strain is very effective. The great advantage of this concept is that the single SMA wire performs dual tasks as both an actuator and a sensor. This has got great importance when the prime objectives are to reduce the overall weight, size and the cost of the actuator system . ArtificIt exhibits robust performance with a small tolerance and can operate without being affected by ambient temperature. In the investigation, the authors have suggested an innovative way to calculate the resistance to determine the change in length of a SMA wire. By doing this, it is possible to measure both the voltage across the entire NiTi wire and that of the fixed-length segment. These two voltages provide direct change in length of the SMA wire. This kind of sensing is used in the feedback control in unknown ambient and loading conditions. This technique is called dual measurement for self-sensing of displacement by resistance change . A self-The sensing of displacement and stiffness without an external sensor is well described in . The worA new mathematical function/model of stiffness that shows the hysteresis characteristics between the stiffness and temperature has been developed and its hysteresis characteristics between the stiffness and temperature is verified by experimental data . It has As the SMA is a highly nonlinear element, its displacement/contraction changes nonlinearly with temperature. It is affected by many electrical and mechanical parameters. Hence, the modeling of the displacement in SMA spring actuator is important. The neural network is the best tool that can easily map one property with others. Therefore, the displacement of the SMA spring can be modeled by ANN and successfully verified by experimental data . In thisIt is known that the Support Vector Regression (SVR) technique is normally applied to forecast the tangential displacement of cement concrete dam. Thus, in general, it is tested and verified using Pearson correlation coefficient, mean absolute error (MAE) and mean squared error (MSE) with experimental data . The impIn research article , the oveAs evident from the literature survey, the self-sensing during actuation of SMA spring is very useful to understand the relationship between the stiffness change and force, displacement, and strain corresponding change in the electrical resistance during the phase transformation. However, so far, a comprehensive study considering several effects such as actuation current, excitation frequencies and duty cycles has not been reported yet. Consequently, the technical novelties of this study are summarized as follows: (a) achievement of the self-sensing behaviour of SMA spring during variable stiffness actuation by experimentation, (b) development of a data driven model of self-sensing variable stiffness actuation based on Support Vector Machine (SVM) regression and nonlinear regression methods, by availing the experimental data, (c) investigation the effect of different excitation/activation currents, switching/excitation frequencies and duty cycle, (d) evaluation of the cycles and pre-stresses on the stiffness-resistance characteristics during the heating cycle of SMA spring by employing SVM algorithm as a soft sensor. From the aspects of the technical novelties, several new characteristics which are significant for the development of self-sensing are found. Some of new findings are given as follows: (i) it is identified that the characteristics of the self-sensing actuation (SSA) are influenced by activation currents, switching/excitation frequencies, duty cycles and pre-stresses, (ii) both SVM regression and nonlinear regression models are acceptable to measure the self-sensing stiffness in SMC actuator during variable stiffness actuation which is experimentally validated with its significance, (iii) it is found that the electrical resistance for all factors is almost in linear relation with the stiffness of SMC during the heating cycle with a meagre hysteresis, (iv) the resistance of SMC is low under the austenite and high under the martensite phases. It is noted here that compact design of a self-sensing SMA spring/wire actuator device is feasible with the aid of the results achieved in this work. This paper is organized as follows. After clearly describing the research motivation, literature survey and the technical contributions of this study, an experimental facility and measurement method are presented in The study to self-sense the stiffness of the SMA spring during variable stiffness actuation from resistance measurement is highly significant with respect to the quality of device . This experimental study is used to validate Support Vector Machine Regression and Nonlinear Regression model and realized by MATLAB 2020b software. A set-up to run the tests is designed and fabricated and shown in Voltage signals from different sensors are converted into proper level (0 to 10 V) through signal conditioning, so that it is compatible with the data acquisition system and stored in the personal computer’s memory. The experimental modeling and analysis of self-sensing of the stiffness in the Shape Memory Coil during variable stiffness actuation is performed in the following ways. In the first set of experiments, activation currents  are varied by D.C. regulated power supply and by keeping voltage constant. Then different sensor voltages, the voltage across rheostat and voltage across the Shape Memory Coil are recorded in the memory of personal computer via DAQ card through repetitive cycle of switching of the power transistor. This procedure is repeated for aforementioned activation currents by keeping the switching frequency, duty cycle and pre-stress constant. The recorded information is used to predict stiffness by nonlinear regression modeling and validation. It is presented in detail in The recorded instantaneous value is used to determine the resistance and stiffness properties of SMC by use of Equations (6) and (7) for the aforementioned activation currents. In the second set of experiments, the switching frequency of the power transistor is changed e.g., 10 mHz, 20 mHz and 30 mHz by ensuring that all other parameters such as activation current, pre-stress on the SMC and duty cycle of the switching frequency are constant. The instantaneous values of the SMA spring’s properties in terms of voltages is continuously recorded in the memory of personal computer via DAQ card. These recorded voltages are converted into proper units of the properties of SMC such as force, displacement, temperature, resistance, and stiffness.Also, the experimental modeling and validations are explained in detail in i is varying from 0 to n and n is number of training data points. The goal is to find the function which map the relation between The regression is statistical tool to model and analyze the relation between one or more than one independent variable and a dependent variable to produce a particular outcome. In other words, the basic idea is to approximate the functional relation between set of independent variable and dependent variable by minimizing risk function which will use prediction error. Kernel: A lower dimensional data set is mapped into a higher dimensional data set using the Kernel function. In essence, a hyperplane is a line that will enable us to estimate a continuous value or aim. Boundary Line: In SVM, a margin is created by two lines other than the hyperplane. The Support Vectors may be on or outside the boundary lines. The experimental data recorded for resistance and stiffness of shape memory coil are denoted as w is weight vector, b is bias and φ(x) is high dimensional data space. The weight vector and bias can be determined from risk function and as,The one dimensional and multidimensional SVR problem is defined asC is regularization constant. The insensitive loss function is defined as,The ary line .(7)fXi,φ(x) is high dimensional data space.jth order polynomial model in one variable is given by,β0, β1,β2,..., iβ, and ……i = 1, 2, 3,…, j are the effect parameters and ε an error.Most self-sensing actuation model literature have related SMA in terms of the displacement/strain with self-sensed electrical resistance. To represent the stiffness of SMA in terms of its electrical resistance, there is a need to establish an appropriate and reliable stiffness-resistance model. To find the appropriate mathematical model that expresses the relationship between dependent variable (stiffness) and the independent variable (resistance), a data driven model is used; it is a parallel to mathematical model with self-sensing characteristics i.e., sensing under actuation. So, the data driven model is a function of the independent variable involving one or more coefficients. The nonlinear regression model with continuous one-to-one mapping is efficient to describe the relation between the dependent variable (stiffness) and independent variable (resistance). The Nonlinear Regression model is preferred as the modeling of the self-sensing actuation phenomenon  is not as complex as that of modeling the basic phenomenon of SMA, the thermo-mechanical phase transformation. Also, the Nonlinear Regression model can be used to approximate complex nonlinear phenomena and then the relationship is curvilinear. The The nonlinear regression of sufficiently high degree can always be found that provides a good fit for data. A good strategy should be used to choose the order of an approximate polynomial; keep the order increasing until t-test for the highest order term is non-significant. It is called the forward selection procedure to fit the model with experimental data. Also, goodness of fit statistics is used to find the best polynomial. MATLAB function “polyfit” is used to obtain coefficients of the polynomials . Data frSMAR is the resistance of the SMA spring (Ω), SMAV is the voltage of the SMA spring (V), R is the known resistance (Ω), SV is the bias voltage of the MOSFET (V),The influence of different factors such as activation current, excitation frequency, duty cycle, and pre-stress on self-sensing characteristics are observed and presented in this section. The influence of different factors such as activation current, excitation frequency, duty cycle, and pre-stress on self-sensing characteristics are observed and presented in this section. The SMA’s electrical resistance is sensitive to compositions, transformation path and heat treatment . During k is the instantaneous stiffness of the SMA spring (N/m), G is the instantaneous shear modulus of the SMC (N/m2), d is the wire diameter of the SMC (m), and D is the coil diameter of the SMC (m). The instantaneous value of G is calculated from force and displacement measurements using transducers.d”. The force-displacement relationship and cooling performance associated to the actuation frequency are most sensitively influenced by the wire diameter “d”, which is a dominant design parameter of the SMA coil spring actuator. The SMA coil spring actuator has the smallest material mass and the quickest cooling time when the wire diameter is the smallest.The first step in the design process is to choose the smallest wire diameter, or “D”. When the shear strain reaches the predetermined value, which is adjusted to be slightly greater than the wire diameter “d”, the force is calculated. If “D” is tiny, the shear stain does not reach its maximum value before the force surpasses the intended value. If so, the calculation is redone with a larger “D” The iteration ends and the “D” value at the last step is the maximum coil diameter if the force reaches the required value at the maximum shear strain while “D” is growing. With the desired stroke and the single coil stroke at the necessary loading condition, the coil number “n” is calculated. The displacement gap between the martensite and austenite models is used to determine the single coil stroke. The desired actuation stroke value divided by the single coil stroke yields the coil number “n”.Iterative calculations are used to determine the coil diameter “So, the initial value of displacement of SMC is assumed to be zero and it is set to zero in the transducers and recorded in the personal computer. This modeling is aimed at the self-sensing phenomenon when the SMA is under variable stiffness actuation i.e., not particularly on the modeling of the basic actuation or shape memory effect or phase transformation of SMA. Moreover, the study is based on the SMA being activated by joule heating, whereby it is controlled by different electrical parameters such as current, frequency and duty cycle and, pre-stress. The change in resistance corresponding to the change in stiffness is determined during the heating phase, and the relation is extracted as a nonlinear regression model and validated by different metrics through experimentation. The model is valid for the joule heating current of 0.7 A to 1.2 A. In the first set of experiments, the SMA spring is electrically heated by a 1/100 Hz square wave signal with a constant duty cycle and constant pre-stress. The current of the heating signal is varied, and the data recorded. The current/electrical power affects both the stiffness and resistance of the SMA spring actuator. The effect of changing current is clearly seen in k is the instantaneous stiffness in N/m and “RSMA” is the resistance in ohm of the SMA spring actuator. Three cases are considered to present the data uniformly corresponding to each effect in parameter variations like current, frequency, duty cycle and pre-stress. The Nonlinear Regression model is used to represent self-sensing actuation in particular to relate stiffness with electrical resistance. The big and unusual coefficient of the nonlinear regression model can be reduced by normalizing stiffness and resistance data. The modeled and experimental self-sensing of stiffness of the Shape Memory Coil during variable stiffness actuation agree in terms of quality as The mathematical Nonlinear regression model between stiffness and resistance during the austenite phase is estimated from the experimental data.The values of metrics of the model comparison for In the second set of experiments, the Shape Memory Coil is electrically heated over a fixed current of 1.2 A of different frequencies  of square wave signal with fixed duty cycle and pre-stress (pre-tension). The self-sensing of stiffness is modeled during the heating cycle only. Both the curves almost agree with each other . The experimental results validated the self-sensing of stiffness of the SMC actuator by measurement of resistance during the heating cycle (austenite phase). The modeled and experimental self-sensing of stiffness of the SMC during variable stiffness actuation agree in terms of quality because ® program, the characteristics for different duty cycles , at a constant current of 1.5 A and constant frequency of 20 mHz showed that they almost agree with each other. The nonlinear regression model of stiffness at different duty cycles with resistance change as independent variable are as follows:An SVR and Nonlinear regression model is developed to understand the effect of duty cycle on stiffness-resistance characteristics. The comparison of experimental and curve-fitted model is simulated by the MATLABThe effect of duty cycle on stiffness-resistance characteristics are linear and useful in controlling stiffness effectively. The comparison between the experimental and modeled characteristics at different duty cycles are presented in The effect of pre-stress on stiffness-resistance characteristics is developed as mathematical models for different pre-stresses  at a constant current of 1.2 A and constant frequency of 10 mHz. The nonlinear regression model of stiffness at different duty cycles with resistance change as independent variable are as follows:The comparison between experimental and modeled characteristics at different pre-stresses found in With the help of four sets of experiments, the stiffness-resistance characteristics of the SMC actuator are analyzed to explore its self-sensing capability. Data recorded from the first set of experiments are used to plot and analyze. Resistance response to the heating cycle is plotted for 50 s; it found that as activation current increased, resistance decreased and that the change of resistance decreased over a period of time as shown in Data recorded from the second set of experiments are used to plot and analyze. Self-sensing actuation characteristics of the SMA spring actuator is obtained for varied frequencies from 10 mHz to 30 mHz keeping activation current constant at 1.2 A; Stiffness is determined and plotted as shown in At a constant current (1.2 A) passing through the SMA spring and constant frequency (10 mHz) of excitation current, stiffness is determined for pre-stress which is varied from 100 g to 200 g. An increase in pre-stress beyond 200 g would not allow complete contraction (bias force is higher) and would not completely deform below 100 g for the requirement of restraining/pulling force . It is lSimilarly, for different duty cycles  at 1.5 A and 20 mHz, the resistance response, stiffness response and stiffness—Resistance characteristics of the SMA spring actuator are obtained and presented in Experimental data is collected for repeated cycles (3 consecutive cycles), and the responses are similar/not with much deviation. One set of data is used for the plotting and analysis of the stiffness-resistance characteristics, the usable form of the data is through the use of an average function and normalization; the other set of data are used for the validation. The highest change in resistance corresponding to the change of stiffness with regard to the effect of the influencing factors like activation current, excitation frequency, pre-stress and duty cycle are presented in 2). The developed SVM model showed an excellent result of prediction in comparison with the nonlinear regression model and experimental data. The experimental analysis has proved that the stiffness of the SMA is sensed from its resistance change. While the stiffness is changed due to different activation currents/joule heating, excitation frequencies, pre-stresses and duty cycles, the stiffness of the SMC is successfully determined as the variable stiffness actuator. Among many new findings from this work, the most interesting result is that the stiffness of the SMA spring can be measured without knowing the activation current and initial geometry or configurations of the SMC. This is possible from the realization of both SVR and nonlinear regression models of the stiffness using the electrical resistance of the SMC during austenite phase transformation. The responses achieved from two models are compared to the experimental response showing both models would harness the self-sensing capability of the SMC actuator. In addition, it has found from this work that the effect of frequency and duty cycle is more linear when compared to the other two parameters of current and pre-stresses. It has been concluded from this work that from the practical view of point, the self-sensing of stiffness of SMC can reduce the number of sensors for making application systems associated with shape memory alloys. For example, one stiffness sensor can use for two sensors of force and displacement. Therefore, it is self-explanatory justifying that the self-sensing technique gives birth to sensor-less control systems which are relatively cost effective, and hence the overall system becomes compact in comparison with traditional control systems having dedicated many sensors. Therefore, it is expected that the proposed self-sensing variable stiffness actuation method can be applicable to many control systems including the grasping force of robot grippers, surgical SMA wire of biomedical sciences, vibration and dynamic motion control of flexible structures in aeronautical fields such as morphing control and health monitoring control system using SMA wires Associated magnetic coils. It is finally remarked that some benefits achieved from this work will be demonstrated by applying to robot gripper systems in near future. In this work, an experimental facility is developed to determine the electrical resistance of the Shape Memory Coil (SMC) actuator that is biased by a tensile steel spring under self-sensing variable stiffness actuation. The SVM regression model is constructed based on experimental data (Expert Knowledge) and provided excellent performances. The performance of the SVM regressor model is verified by a correlation coefficient, mean square error (MSE), root mean square error (RMSE) and goodness of fit (R


Pythia8 event generator. We show how colour reconnection models and hadronization mechanisms influence such angular correlations and in particular address the effect of gluons on the baryon production mechanism in the Lund string fragmentation model. We conclude by discussing the new theoretical ideas in comparison with the ALICE We present the results from our investigation of angular correlations between baryons pairs in the  One of the research interests in particle physics is understanding the production mechanism and the spatial distribution of particles produced in high-energy particle collisions. This can be studied in various ways in colliders, e.g., by measuring single particle distributions as a function of one or more variables or looking at correlations between particles, for different species of particles. Using phenomenological models we can then use these measurements to gain theoretical insights into the underlying particle production mechanisms.In this work, we address a long-standing open question about the angular correlations of pairs of the produced hadrons. A two-particle correlation function provides information regarding the production of another particle near the first particle. It is often studied as a function of relative pseudorapidity  [Pythia. All these steps can influence the production rate of different hadrons, and correlations of the hadrons in the simulation results.Phenomenological models like s (MPIs) , and theion (CR) , 9 modelion (CR)  in PythiPythia8 is able to reproduce the angular correlations for the same- and opposite-sign meson pairs fairly well, we do not discuss the mesons’ angular correlations in the rest of the paper. Instead, our investigation is focused on the angular correlations of the same- and opposite-sign baryon pairs. We also keep our results limited to protons while discussing various theoretical aspects.For simplicity, we keep our investigation limited to Pythia8 implementation of the Lund string fragmentation model. Special attention is given to the role of gluons and how they may affect the production of baryons. This is followed by Sect. Pythia using the QCD-inspired colour reconnection model. In Sect. In the following, we will start in Sect. Pythia, from multiple scatterings, initial- and final-state showers, the tracing of colour connections between partons is done using a leading-colour  jets contribute to the baryon angular correlations, it is relevant to scrutinise the baryon production in a Lund string with gluon “kinks” a bit closer.Pythia is that the creation of a virtual The general idea behind the popcorn model used in entclass1pt{minimaq must be longitudinal towards the quark end of the string and vice versa for the q and For the mentclass2pt{minimq and This picture works nicely for a straight string. But if there is a gluon along a string, the picture changes. In Fig. Pythia8, there is no special treatment of baryon production close to such gluon kinks. From the description of the popcorn model above, however, it is clear that for a non-breaking virtual In the current implementation of string fragmentation in Since we have here shown that gluons are important for the azimuthal correlations between baryons we will in Sect. Pythia. In some cases non-trivial colour topologies may arise prior to the string fragmentation stage, e.g., from the treatment of remnants in hadronic collisions, or when looking at baryon number violating BSM processes. In the MPI machinery, it is not uncommon that two  quarks are taken from a proton, leaving a remnant in a colour-triplet state. Similarly, baryon number violating processes may decay a colour-triplet particle into two anti-triplet particles. In both cases, we may obtain colour-singlet string systems connecting three quarks (or three anti-quarks) in a so-called string junction topology [Pythia8 is able to hadronise such systems, in a process that always will produce a net baryon number.The popcorn and di-quark models are not the only way of obtaining baryons in topology . Pythia8Pythia8, using the so-called QCD colour reconnection model [We will not be concerned with BSM here, and the junctions formed in the MPI remnant treatment mainly affects baryons in the far forward or backward regions of rapidity. There is, however, another way of creating junctions available in on model .Pythia has a default CR model, which is based on MPIs [CR models re-arrange the colour connections of the colour dipoles produced after MPIs and parton showers. The primary objective of CR is to reduce the net string length so that the model can reproduce the charged particles’ multiplicity and the observed enhancement in  on MPIs , where tPythia8 has an alternative model, the QCDCR model [swing. In (b) we instead have the situation where the two (anti-) colour ends together are in an anti-triplet (triplet) state and can reconnect to two junction systems connected by a dipole. Finally, in (c) the (anti-) colour ends of three dipoles together form a colour singlet and can reconnect into a separate (anti-) junction system. In each case only reconnections that reduce the overall string lengthsCR model , which fThe number of junctions in It should be noted that the two connected junctions can be separated by a long dipole or a chain of multiple dipoles that are reconnected and can separate the junctions by a large rapidity span. For example, the junction baryon and anti-baryon produced in Fig. Pythia8 at small angles. Fermi–Dirac effects would decrease the correlation at small angles for identical baryons, but again the effect is expected to be small.There are many potential final-state effects that may affect correlations between hadrons produced in the string fragmentation. The Lund group has studied several such models, e.g., a model for Fermi–Dirac correlations , the so-entclass1pt{minimaInstead, we will focus on the model for hadronic rescattering , 20. By Pythia, and in this section, we will confront these ideas with data. It can be noted that we have also tested varying standard string fragmentation parameters, such as flavour ratios, di-quark production rate, spin ratios of the di-quarks, and Pythia.So far we have presented a set of ideas that may affect baryon correlations in Pythia8 to produce at most one di-quark breakup per string, it was possible to understand the correlations found in data. Such an artificial change in the behaviour of the string fragmentation is of course not a satisfactory solution, but it gives us hints as to what is needed. We will therefore concentrate on reducing the number of di-quark breakups in strings with (semi-) hard gluons, but also to introduce alternative baryon production mechanisms that do not exhibit the correlations found in string fragmentation. In addition, we will also consider final state effects from hadronic rescattering.In Ref. , the maiPythia version 8.306 to generate In all simulations, we have used the he Rivet  routine Pythia8.We begin with the QCDCR model, which introduces a completely new way of producing baryons. We have used the so-called “mode-0” tune presented in , with noThe choice of the CR model does not, however, improve the angular (anti-) correlations for the same-sign baryon pairs. The effect is rather a general reduction in correlations, which is expected since the model will produce additional baryons with fewer correlations.The reduction is more clearly seen for the opposite-sign baryon pairs, where QCDCR model reduces the amplitude of the baryon-antibaryon pair correlations near It should be noted that we have also studied the effects in meson correlations (which are not shown here) but found no significant effect of the choice of CR model there.Since the QCDCR shows significant improvement in the angular correlations of the opposite-sign baryon pairs, we will in the following use the QCDCR as our base-line set-up when adding other modifications.Pythia, implementing The produced hadrons can interact with nearby hadrons via elastic or inelastic scattering. A model for hadronic rescattering  was receThe reason for this is somewhat non-trivial, and is related to the annihilation of baryons–anti-baryon pairs in the rescattering. As explained in Sect. mentclass2pt{minimThere is currently no proper implementation for the possible suppression of baryon production in string fragmentation close to gluon kinks in the popcorn mechanism discussed in Sect. Pythia8 [We have decided to constrain the baryon production using the  Pythia8 , which aC and the next breakup is a diquark–anti-diquark breakup in region B, we veto the baryon to be produced, and tell Pythia to try another breakup instead. It should be noted that the Lund string fragmentation model is left–right symmetric, and if we go from the other (B the following C producing the same baryon from a kinky string piece, is not vetoed. The reason for implementing it in this way is technical, but effectively it will result in a suppression of baryons produced around a string corner with a factor of 0.5.In our crude implementation, we veto any baryon produced in a diquark breakup if the previous breakup was in a different string region. As an example consider the case in Fig. Pythia8 including QCDCR. As expected the jet peak for baryon pairs is reduced, and for In Fig. It should be noted that our toy model will reduce the overall number of baryons produced in general, even in Pythia and the accumulated changes of all three models investigated her: QCDCR, hadronic rescattering and the vetoing of baryons close to gluon kinks. We see that the reproduction of the ALICE date is far from perfect if the models are added, but there is a clear improvement over the default Pythia8. The jet peak is reduced both for As a final result, we show in Fig. Pythia in reproducing these experimental results. We found that two already existing models in Pythia8, the QCDCR model and the hadronic rescattering model, have a significant effect on the correlations, and adding these to the default Pythia8 improves the description of data significantly.We have shown that the observed anti-correlation for the same-sign baryon pairs in the ALICE experiment for Pythia is able to reproduce the angular correlation distribution for the opposite-sign baryon pairs.The QCDCR model produces additional baryons due to junction systems forming as a part of the reconnections of the colour dipoles. Such junction baryons are much less correlated than those produced in string fragmentation. The string connecting two junctions can produce multiple hadrons between the two junction baryons, unlike the popcorn baryons, which are separated by only one meson in between. We show that it visibly reduces the correlations between the opposite-sign baryon pairs in the jet peak near Pythia results, it is not sufficient. Adding the hadronic scattering model, we found that the effect of annihilation of baryon–anti-baryon in jets with more than one baryon–anti-baryon pair is quite significant, while if there is only one pair, there is typically no annihilation. This gives a further reduction of the jet peak for same-sign baryons, while the effects on unlike-sign correlations are small. Still, the jet peak for same-sign baryons in Pythia needs to be further reduced in order to reproduce data.The anti-correlations in the same-sign baryon pairs are rather complex results. Although the QCDCR model improves the Pythia reproduce data, by forcibly forbidding more than one baryon–anti-baryon pair to be produced in a string. This effectively removed the jet peak in the same-sign baryon correlations, leaving only the anti-correlation in the away-side ridge. Here we instead propose a more physical mechanism, where baryon production close to gluon kinks in a string is suppressed. The motivation for this comes from the popcorn model of baryon production in a string. Here an extra non-breaking virtual The authors in  managed Pythia8, but it is still likely that the jet peak for same-sign baryon correlations would be reduced. Whether it will be reduced enough to reproduce data remains to be seen.Since the jet peak around We believe that the baryon suppression near gluon kinks will affect the baryon-to-meson ratios in Finally, we note that there are other independent measurements that could verify our hypothesis of suppressed baryon production close to gluons. One obvious example is to compare the baryon-to-meson ratio inside a gluon jet to that of a quark jet, which could be done by comparing inclusive jets to jets produced together with hard photons. We are not aware of any study where the jet substructure has been studied for identified hadrons, but we would certainly like encourage our experimental colleagues to pursue such measurements at the LHC.