Arabidopsis thaliana), accumulate starch in the light and remobilize it to support maintenance and growth at night. Starch synthesis and degradation are usually viewed as temporally separate processes. Recently, we reported that starch is also degraded in the light. Degradation rates are generally low early in the day but rise with time. Here, we show that the rate of degradation in the light depends on time relative to dawn rather than dusk. We also show that degradation in the light is inhibited by trehalose 6-phosphate, a signal for sucrose availability. The observed responses of degradation in the light can be simulated by a skeletal model in which the rate of degradation is a function of starch content divided by time remaining until dawn. The fit is improved by extension to include feedback inhibition of starch degradation by trehalose 6-phosphate. We also investigate possible functions of simultaneous starch synthesis and degradation in the light, using empirically parameterized models and experimental approaches. The idea that this cycle buffers growth against falling rates of photosynthesis at twilight is supported by data showing that rates of protein and cell wall synthesis remain high during a simulated dusk twilight. Degradation of starch in the light may also counter over-accumulation of starch in long photoperiods and stabilize signaling around dusk. We conclude that starch degradation in the light is regulated by mechanisms similar to those that operate at night and is important for stabilizing carbon availability and signaling, thus optimizing growth in natural light conditions.Many plants, including Arabidopsis ( Starch degradation in the light is regulated by similar mechanisms to those operating at night, stabilizing carbon availability, and thereby optimizing growth in natural light conditions Arabidopsis thaliana) accumulate a substantial part of their fixed carbon (C) as starch in the light to support maintenance and growth at night . This model explains many observations, especially the robust timing of starch degradation to the coming dawn in the face of sudden perturbations (see above). It has also been proposed that starch turnover is directly or indirectly regulated by metabolic inputs that alter circadian clock gene expression and clock phase to maintain sugar homeostasis  before slowing and plateauing from about ZT14 onwards , the absTo examine further whether the onset of starch degradation is a function of time after dawn rather than anticipated time of dusk, we measured the impact of decreases in irradiance imposed at different times on the accumulation of starch. We showed previously for plants grown in 12-h days that the effect of a decrease in irradiance depended on when the decrease was imposed. A decrease at midday (ZT6) resulted in a reduced starch accumulation rate commensurate with the reduction in photosynthesis, whereas the same decrease after ZT9 resulted in a plateauing or even a decline in starch levels . These p−2 s−1 for short-day plants and from 80 to 50 µmol m−2 s−1 for long-day plants. These decreases, which were previously shown to cause a decline in the rate of photosynthesis of about 60% , but not relative to subjective dusk. After irradiance decreases at ZT4 in short-day plants  and at Z13CO2. The total amount of starch that accumulated during the pulse was compared with the amount of 13C that accumulated in starch. Similar increases in 13C-starch and total starch (i.e. 13C- plus 12C-starch) during the pulse would be consistent with the idea that preexisting starch is not being degraded. A larger increase in 13C-starch than in total starch (i.e. a decrease in the amount of 12C starch) would show that some of the preexisting starch is being degraded. This is plausible given the recent demonstration that Arabidopsis leaf starch granules grow anisotropically  divided by the remaining time to dawn (T). The finding that the onset of rapid starch degradation in the light depends on time relative to dawn, rather than dusk, prompted us to ask if a skeletal form of the arithmetic division equation can also be used to predict changes in starch content after transfer to continuous light. The following data analysis is independent of the molecular models developed by Rd = S/T that describes the observed responses at night can also be used to predict temporal changes in starch content in the light. With time in the light, starch content rises, while the time until the next dawn decreases. This should give rise to an increasing rate of starch degradation with time, if the model holds in these conditions. Assuming that the rate of starch synthesis is constant in the light (see below for data supporting this assumption), an increasing rate of degradation with time could result in a progressive shift from net starch accumulation to net starch mobilization. These expectations are in line with our measurements of starch and maltose, which point to a gradual rise in the starch degradation rate with time in the light (decreasing time until the next dawn) and are thus qualitatively consistent with the response predicted by the arithmetic division equation.The arithmetic division model  was form−2 s−1. We collated data because there is experiment-to-experiment variation in the response of starch content in continuous light; this may be due partly to biological noise and to differences in the metabolic status of the plants between experiments. One such experiment with Arabidopsis accessions Col-0 and Ws-2 is shown in the left-hand display of sd) of that in the subjective day, whereas in continuous low light the rate slowed to −15% ± 22% of that during the subjective day . The difference between the two responses was significant  .To provide a more rigorous test of the ability of the skeletal arithmetic division equation to predict the observed changes in starch content in the light, we collated data on starch content from many experiments in which plants were transferred at dawn to continuous light at either 160 or 90 µmol mRd = S/T)  predicted the observed gradual slowing down of net starch accumulation with time in the light  gene under the control of the ethanol-inducible ALCOHOL DEHYDROGENASE REGULATOR (AlcR) promoter to show that elevation of Tre6P during the night results in a reduced rate of starch degradation (TPS29.2) to test whether elevation of Tre6P also inhibits starch degradation in the light. As a control we used AlcR lines that express the AlcR ethanol-binding transcription factor but do not carry the bacterial tps gene  at ZT0, then sprayed with ethanol at ZT10 to induce AlcR protein and (in the case of TPS29.2) Tre6P production. Levels of Tre6P were significantly higher in TPS29.2 than AlcR by ZT12, and up to 2.5-fold higher between ZT15 and ZT24  or between ZT14 and ZT20 (after induction with ethanol). Between ZT2 and ZT8, total starch and 13C-starch accumulated in similar amounts in both lines  and UDP-glucose , as expected if the C in these metabolites were derived in part from degradation of preformed  starch  falls, that is (Rd* = (S/T)(1-A), We modified the skeletal arithmetic division equation (Model 1) to include a term that links the capacity for starch degradation to the rate of photosynthesis, such that starch degradation is inhibited during photosynthesis but is progressively activated as photosynthesis rate  the rate of accumulation of 13C in glucose in the cell wall fraction, representing synthesis of cellulose and the backbone of hemicellulose and (2) the rate of accumulation of 13C in protein. Most free amino acids show slow and incomplete labeling, which complicates or prevents their use for estimation of protein synthesis rates  to determine 13C enrichment in glucose in the cell wall polysaccharides, in free Ala and Ser, and in Ala and Ser in protein.Plants were grown in an 18-h photoperiod in a conventional step  light regime at 142 µmol m m−2 s−1 . This de13C enrichment in cell wall glucose  or, as a control, in a 6-h photoperiod at 160 µmol m−2 s−1  or transferred at dawn to 90 µmol m−2 s−1  suddenly increases. This apparent discrepancy was investigated in material from an experiment in −2 s−1 were transferred to continuous low light (90 µmol m−2 s−1) and sampled until ZT33 (9 h after the first subjective dawn). As reported in A and ADPGlc content remained high throughout the continuous light treatment, starch content plateaued by ZT15, showed a slight decline between ZT15–ZT24 followed by a slight rise between ZT24–ZT33 , and sucrose, glucose, and Glc6P levels rose in the subjective night and declined after ZT24 . We used this equation to simulate the kinetics of starch accumulation after transfer to continuous light (1-A) that included a term for feedback inhibition by Tre6P simulated the response of starch accumulation after transfer to continuous high or low irradiance  [L.] Heynh. Col-0 was grown on soil in controlled environment rooms as in −2 s−1, 75% relative humidity). A list of all experiments with the experimental design, which data displays was generated from them and the location of the accompanying data sets is provided in Arabidopsis (N-methyl-N-trimethylsilyltrifluoroacetamide followed by gas chromatography time-of-flight mass spectrometry (GC–TOF–MS) analysis using conditions and settings as in Rosettes were harvested under ambient light conditions and frozen immediately in liquid nitrogen. Sugars and starch were extracted and measured enzymatically . Tre6P a2 from the soil into the chamber air space.Gas exchange was measured using a LI‐6400XT Portable Photosynthesis System fitted with the 6400‐17 Whole Plant Arabidopsis Chamber  and testing that no starch remained, the pellet was chemically digested and the released glucose determined by GC–TOF–MS as described . The same procedure was followed for imposition of a decrease in light intensity at ZT6 and ZT10, from 105 to 65 μmol quanta m−2 s−1.Plants were grown in a 12-h photoperiod at 150-μmol quanta m−2 s−1 leads to a ∼50% inhibition of photosynthesis. The rate of starch synthesis, Rs, was assumed to be constant with time in the light , the rate of starch degradation, Rd*, =  (S/T)(1-A) where A is a nominal term for a rate-of-photosynthesis-dependent inhibition of starch degradation. Based on Rs – Rd (Model 1) or Rs − Rd* (Model 2). In panels D, H, and F where the response to a halving of light intensity was simulated, the rates of photosynthesis and starch synthesis were assumed to decrease in proportion with light intensity.The change in starch content with time in the light was predicted as in the legend of toperiod , or aftetoperiod . Rs was radiance . Starch radiance  in the sRd = S/T where S is the amount of starch and T is the time until anticipated dawn (Rd = S/T), Model 2: Rd* = (S/T)(1-A), where A is a nominal term for a rate of photosynthesis-dependent inhibition of starch degradation, set as proportional to light intensity, L. The calculations were applied iteratively over 1-h intervals. In Models 1 and 2, starch degradation in the light was allowed between ZT15 and ZT18 and between ZT0 and ZT3 . In Models 1 and 2, starch content in the light was corrected for starch lost by degradation. In all calculations, it was assumed that the clock is entrained to 3 h after first light, as is seen in plants growing in long days . Starch degradation is negligible in the first hours of the light period in short-day conditions . Maltose, starch, ADPGlc, sucrose, glucose, and Glc6P content and A, after transferring plants to continuous light  . Total starch content and 13C enrichment in starch in long day-grown plants maintained at growth irradiance throughout their growth and on the day of the experiment, and pulsed with 13CO2 for short periods in the first hours after dawn  grown in short-day conditions and pulsed with 13CO2 and harvested in the first hours after dawn . Simulation of the impact of a sudden decrease in irradiance on C availability for maintenance and growth during twilight in short-day-grown plants (background to kiac162_Supplementary_DataClick here for additional data file.


Aim: The aim of this paper is to perform a retrospective assessment of the clinical performance of the complete oral rehabilitation of patients with bruxism treated with implants and teeth-supported veneered and non-veneered monolithic zirconia restorations with increased occlusal vertical dimension. Methods: In this retrospective follow-up study, 16 bruxer patients, mean age 59.5 ± 14.9 years, were treated with 152 veneered and 229 non-veneered monolithic zirconia and followed for a mean of 58.8 ± 18.8 months (range 1–8 years). The patients were examined clinically and radiographically, annually. Clinical data were extracted from the medical records. In the recall appointments, modified California Dental Association (CDA) criteria were used to evaluate the restorations. Implant and restoration survival and success rates were recorded and analyzed. Results: The cumulative survival rates of implants and restorations were 97.7% and 97.6%, respectively. Nine restorations were replaced: three due to horizontal tooth fractures, two because of implant failure and four had secondary caries. A total of 43 biologic and technical complications were recorded. In the veneered group, the predominant complication was minor veneer chipping (16.4%), which required polishing only (grade 1). In the non-veneered group, the main complication was open proximal contacts between the implant restorations and adjacent teeth (14.5%). Conclusions: The survival rates of restorations and implants in patients with bruxism are excellent, even though veneered zirconia restoration exhibited a high rate of minor veneer chipping, which required polishing only. The biologic complication of fractured single-tooth abutment may occur. Bruxism, as defined by Lobbezoo et al. , is a reHigh-strength monolithic zirconia ceramic restorations have become the preferred treatment for patients with bruxism . An overIn a recent study, comparing monolithic zirconia restorations in bruxer versus non-bruxer patients, no significant differences were found between the two groups regarding overall complications and survival rates . HoweverIn a retrospective study on implant-supported, full-arch, fixed dental prostheses, with a mean follow-up of 10 years, Chrcanovic et al.  showed tSevere bruxism is often associated with extensive attrition and erosion that necessitates rehabilitation of complete dentition with an increased vertical dimension of occlusion (VDO). The question remains whether this change in VDO through extensive rehabilitation, especially in bruxer patients, causes increased rates of prosthetic complications.In a multicenter retrospective clinical study, Fabbri et al.  found thThe aim of this retrospective study was to make retrospective assessment of the clinical performance of the complete oral rehabilitation of patients with bruxism treated with implants and teeth-supported veneered and non-veneered monolithic zirconia restorations with increased VDO.Electronic records of patients with bruxism, who had been fully rehabilitated with monolithic zirconia restorations via an increased VDO, were screened from the records of the graduate clinic of prosthodontics in the Dental Medicine School and from the private practice of the program director (S.L.), who is an experienced prosthodontist.Patients were included only if they were diagnosed with bruxism according to the international consensus of Lobbezoo et al. , underweThe exclusion criteria were less than 1 year of follow-up, an uncontrolled medical condition, aggressive periodontal condition, incomplete records or unavailability for recall. The patients underwent clinical and radiographic examinations annually. These clinical data, as well as the preoperative data, were extracted from the medical records.For each patient, the diagnostic grading system of “possible,” “probable” or “definite” sleeping or waking bruxism was recorded at the time of arrival, according to the international consensus  and the The detailed study protocol was previously described . The samIf the amount of the increased VDO was within the interocclusal rest space (IORS) of the patient, the temporary acrylic resin restorations were delivered immediately after mock-ups were tested. If the amount of the increased VDO was beyond the range of the IORS of the patient, a removable appliance was used for two months in order to test the adaptability of the new VDO. The temporary restorations were delivered only if no functional complications were reported .After at least two months of normal function with the new VDO, final zirconia restorations were fabricated and delivered on teeth and implants, either as single abutments or short, fixed partial dentures with only one pontic. Implant-supported restorations were either screwed or cemented to the implant abutments.The design of the non-veneered, monolithic zirconia was created using CAD-CAM software according to the manufacturer’s specifications . For the veneered monolithic zirconia design, facial cutback was carried out according to a virtual modification, and a feldspathic ceramic veneer was added to the nonfunctioning facial surfaces.With the delivery of the final restorations, all patients received occlusal splints and were instructed to wear them during sleep.Two examiners (S.L. and H.H.) were trained and independently assessed all radiographic and clinical records of the zirconia restorations and the abutments, implants and teeth. In case there was a disagreement in the rating of a certain restoration, the case was discussed by both examiners until they reached a consensus. Periapical radiographs of each implant and tooth abutment were taken using the long cone technique and were compared with the radiographs taken with the definitive prosthesis. Survival, biological and technical complications of all teeth, implants and prostheses were recorded.For each implant, the following biological findings were recorded: periodontal probing depth, bleeding on probing, and peri-implant suppuration, if present. Implant survival was defined as a functioning implant with no clinical signs of infection, even if bone resorption was identified radiographically. Implant failure was defined as an implant that did not survive .The monolithic zirconia restorations were assessed technically according to the modified California Dental Association (CDA) quality evaluation system for assessing surface, color, shape and marginal integrity ,15,16. PDescriptive statistics of patient age, follow-up time, and VDO increase were carried out using IBM SPSS statistics, version 25.0 . Life table analysis was used to assess implant and zirconia restoration survival. Kaplan–Meier curves were used to estimate the zirconia restoration survival and complication-free rates.The overall pool examined included 18 patients  who met the inclusion criteria. Two did not attend the recall appointment; one did not respond to our invitations, and one was deceased. Therefore, only 16 patients  attended the recall appointment. The participants ranged from 32 to 78 years of age (mean: 59.5 ± 14.9 years) and were followed for a mean observation period of 58.8 ± 18.8 months (range 1–8 years). All patients were diagnosed with “probable bruxism” [No temporomandibular signs or symptoms were reported. Among the 88 implants restored, 19 (21.6%) were in the anterior region and 69 (78.4%) were in the posterior region of the jaws. Fifteen implants (17%) had cement-retained restorations, while the rest (83%) had screw-retained restorations.The patients received restorations of a non-veneered monolithic zirconia design in the mandible and in the posterior quadrants of the maxilla. In the upper anterior crowns , the buccal surfaces were veneered with a feldspathic ceramic material. Therefore, a total of 152 restorations were veneered monolithic zirconia, while the remaining 229 restorations were a non-veneered monolithic zirconia design. In the recall appointment, only seven patients reported using the occlusal splint during sleep .®, Zimmer Biomet, Palm Beach Gardens, FL, USA). Both failed implants were placed as part of the rehabilitation and had no complication or inflammation around them before the rehabilitation. Life table survival analysis concerning implant survival rate is shown in Bleeding on probing was found on one or more teeth/implants in all patients. The periodontal probing depth was between 3 and 5 mm for all teeth and implants, except for two implants that failed during the follow-up period due to peri-implantitis. Thus, the cumulative implant survival rate was 97.7%. One implant, with a cement-retained restoration, failed in a female patient after 5 years  and the other, with a screw-retained restoration, in another female patient, failed after 6 years  in two patients. Patient 2 had two vital fractured premolars  had secondary caries, and their zirconia restorations had to be replaced a–c.The last two failed restorations were of the failed implants. One was replaced with a new implant and a new zirconia restoration was delivered. The other implant was not replaced due to patient refusal.The remaining failures were classified as complications (with/without repair). Among the 43 complications, 27 occurred in the veneered group and 16 in the non-veneered zirconia group .Twenty-five feldspathic porcelain veneers in the anterior restorations (incisors and canines) chipped. All were on the incisal edge and required only polishing (grade 1) .Two minor zirconia fractures were observed in patient 16, which did not require any repair. Patient 7 experienced a retention loss of three cemented implant-supported restorations, and one loss of tooth vitality. The implant-supported restorations were recemented, and the non-vital tooth was treated with a root canal through an opening in the zirconia restoration. Twelve open contacts, affecting eight patients, were observed between implant restorations and the adjacent teeth: two were in the upper anterior region and the others in the posterior quadrants . Eleven Therefore, the implant restoration was removed, and feldspathic porcelain was added in order to achieve better contact with the adjacent tooth.The non-veneered and veneered monolithic zirconia restorations received separate CDA ratings.In the non-veneered monolithic zirconia restorations, the surfaces were excellent (100%) and the color was satisfactory (100%). The shape was excellent in 80% of the restorations, 0.9% were unsatisfactory-irreparable (two restorations were replaced due to implant failures) and 19.1% were satisfactory, including the 10 open contacts between implants and teeth without food impaction. The margins were excellent in 56% of the restorations; 1.7% were unsatisfactory and irreparable due to four secondary caries that needed to be replaced. The rest were rated as satisfactory with slight over-contouring (42.3%). Overall, 2.6% needed to be replaced, and the remaining 97.4% non-veneered monolithic zirconia restorations were evaluated as satisfactory and not in need of any repair or revision .In the veneered monolithic zirconia restorations, the surfaces were excellent in 83.6% and the rest were satisfactory (16.4%) due to the minor veneer chipping, which required only polishing (grade 1). The color was rated as excellent in 40%, whereas 60% were rated as satisfactory. The shape was excellent in 90% of the restorations, 2.0% were unsatisfactory-irreparable (three restorations were replaced due to fractured teeth), 0.6% were unsatisfactory-reparable (one open contact that needed repair) and 7.4% were satisfactory in 7.4%, including the other open contact which did not have impacted food. Half of the restorations had excellent margins, while the rest were slightly over-contoured and rated as satisfactory. Overall, 2% needed to be replaced, 0.6% needed repair and the rest of the veneered monolithic zirconia restorations; 97.4% were evaluated as satisfactory .The results of the Kaplan–Meier survival and complication-free rate analyses of all the zirconia restorations (veneered and non-veneered) are presented in The current study evaluated the clinical performance of the complete oral rehabilitation of patients with bruxism treated by implants and teeth-supported, veneered and non-veneered, monolithic zirconia restorations with an increased VDO after a follow-up of 1 to 8 years. The VDO was increased by 2 to 7 mm (mean 3.3 ± 1.6 mm). All patients adapted completely to the new VDO, without reporting any temporomandibular signs or symptoms throughout the follow-up time.A total of 43 relative failures (complications) were divided between the two groups . In the 0.5) but a major drawback of opacity [In the current study, we used 3Y-TZP, which has high flexural strength (900–1200 MPa) and fracture toughness  after a 1-year follow-up, but there was no statistical difference between the two groups on the distal surfaces. The open proximal contacts in the current study were found in eight patients, six of whom reported not wearing the night guard, while the other two were found in patients who reported using the splint . This is a common finding in other studies, regardless of bruxism. In a systematic review and meta-analysis, Bento et al. reported proximal contact loss rates of 41% with no significant difference between the posterior and anterior regions . This ree splint .The cumulative zirconia restoration survival rate was 97.6% due to nine absolute failures . An impoOther absolute failures were the secondary caries that occurred at the marginal integrity of four zirconia restorations and the two failed implants in the non-veneered group . A 1% inLimitations of the current study include the retrospective design, in which clinical procedures are not standardized as they could be in a prospective follow-up. The small sample size and lack of a control group of non-bruxer patients can also be considered limitations of the present study. Additionally, the diagnosis of bruxism was based only on clinical inspection and self-report, without polysomnography examination. Additional, long-term, randomized, controlled trials with larger sample sizes are needed to assess the effect of bruxism on the survival of complete rehabilitation with monolithic zirconia restorations and increased VDO.Within the limitations of this retrospective study, we conclude that complete oral rehabilitation using monolithic zirconia restorations with an increased VDO in patients with bruxism have high cumulative survival rates of implants and restorations . In the veneered group, the predominant complication was minor veneer chipping (16.4%), which required only polishing (grade 1). In the non-veneered group, the major complication was open contacts between the implant restorations and the adjacent teeth (14.5%). A biologic complication of three fractured teeth, which were replaced with implant-supported restorations occurred in two patients. Therefore, in bruxer patients, when there is minimal tooth structure, connecting two adjacent abutment teeth may be advised.


However, ankle joint stiffness was significantly lower only at maximum speed (p < 0.001). Hip stiffness was significantly higher after walking training than that before training (p < 0.001). In contrast, knee stiffness after training was significantly lower than pre-training stiffness in the same walking condition (p < 0.001). Ankle stiffness differed only at maximum speed, and it was significantly higher than pre-training stiffness (p < 0.001). Walking fatigue appeared to change the mechanical properties of the joint. Remarkably, at the maximum walking velocity in exhaustion, when the load on the hip joint was significantly increased, the knee joint’s stiffness decreased, possibly leading to joint instability that results in exercise injury.The aim of this study was to assess the stiffness of each lower limb joint in healthy persons walking at varying speeds when fatigued. The study included 24 subjects . A Vicon three-dimensional analysis system and a force plate were used to collect lower extremity kinematic and kinetic data from the participants before and after walking training under various walking situations. Least-squares linear regression equations were utilized to evaluate joint stiffness during single-leg support. Three velocities significantly affected the stiffness of the knee and hip joint ( Walking is one of the most popular sports and is performed by a wide range of individuals . It requStudies have demonstrated a dosage connection between physical activity and health benefits . Walkingjoint) is the mechanical force that resists displacement inside the corresponding joints of the lower limbs , which is determined by the physiological structure of the corresponding joints, the mode of motion, and the mechanical characteristics of the joints . Ve. Ve44]. The kinematic data were preprocessed with Vicon Nexus software to capture the whole walking phase, process any missing marker points, and eliminate any inaccurate or duplicated marker points. The initial kinematic and kinetic data were further processed using Visual 3D software , and fourth-order Butterworth low-pass filters with cutoff frequencies of 15 and 50 Hz, respectively, were employed for smoothing . Three-dhip), knee stiffness (kknee), and ankle stiffness (kankle) were calculated during the active work phase by graphing the slope of the linear regression between the bending/extension moment in the sagittal plane of each joint and the bending/extension angle during stance [In this study, we focused on the stiffness variations in the lower extremity joints during single-leg support (between foot flat and heel lift). The lower limb hip stiffness  of these joints is computed to evaluate the data’s linearity; if r2 is more extensive than 0.80, the phase is regarded to have a moment-angle relationship that is extremely close to linear behavior [After measuring the stiffness of each joint, the determinant  was used, followed by a post hoc analysis when a main or interaction effect was detected. The follow-up pairwise comparison alpha level was set to 0.05 divided by the number of comparisons. A Sidak post hoc test for repeated measures analysis of variance (ANOVA) was employed . Furthermore, Pearson product-moment correlation was conducted to estimate the association between walking speed and joint stiffness. Paired t-tests were performed to determine differences in individual joint stiffness at the same walking pace before and after training. SPSS was used for all statistical analyses .2 > 0.80).Participants walked at normal speed, 25% faster, and at maximum speed, most of the analyzed phases had a nearly linear moment–angle relationship . As shown in p < 0.001). Post hoc tests showed that increases in hip angle, moment, and stiffness were significantly different between normal speed and 25% faster speed and between normal speed and maximum speed, and between 25% faster speed, and maximum speed (p < 0.001). In addition, walking speed was significantly and positively correlated with hip stiffness . Using paired t-tests, hip stiffness before and after walking was compared in the three walking conditions . As shown in p < 0.001).In the case of hip joint stiffness, there was an interaction effect between fatigue and velocity (p < 0.001). As shown in p < 0.001). Post hoc tests showed that the increases in knee angle, moment, and stiffness were significantly different between normal speed and 25% faster speed, between normal speed and maximum speed, and between 25% faster speed and maximum speed (p < 0.001). In addition, walking speed was significantly and positively correlated with knee stiffness . Paired t-tests comparing knee stiffness before and after walking in the three walking conditions  showed that knee stiffness after walking training was significantly lower than that before training for all three walking conditions, as shown in p < 0.001).In the case of knee joint stiffness, there was an interaction effect between fatigue and velocity (p > 0.05). As shown in p < 0.001). Post hoc tests showed that increases in ankle angle, moment, and stiffness were significantly different between normal and maximal speed and between 25% faster speed and maximal speed (p < 0.001) but were not statistically significant between normal and 25% faster speed (p > 0.05). Ankle stiffness at maximal speed was significantly lower than at normal speed and at 25% faster speed. In addition, walking speed was significantly negatively correlated with knee stiffness . Paired t-tests comparing ankle stiffness before and after walking in the three walking conditions  showed that ankle stiffness before and after training was statistically significant only at maximum speed and was significantly greater after walking training than before walking training (p < 0.001), while there were no obvious differences in ankle stiffness between the remaining two walking conditions (p > 0.05).In the case of ankle joint stiffness, there was no interaction effect between fatigue and velocity . This occurrence might be linked to such walking features as the lower load on the hip joint while walking and the stronger fatigue resistance of the hip joint supported by vast muscle groups compared to the other two joints [p < 0.001). Prior studies have shown that the knee joint has a high mechanical energy absorption capacity during exercise [Under the same walking conditions, we found that hip stiffness was significantly higher in exhaustion than that before training , the ankle joint stiffness revealed significant variations only at maximum speed, at which it was much lower than it was at the other two speeds. When joint stiffness before and after fatigue was compared under the same walking conditions, it was shown that hip stiffness was much higher than that before training, and knee stiffness was significantly lower than that before training. Walking fatigue appears to modify the mechanical characteristics of the joints, particularly at the maximum walking speed following fatigue, which significantly increases the load on the hip joint, according to the findings of this study. The knee joint’s lower stiffness revealed unstable features. Ankle joint stiffness differed before and after exhaustion only at the quickest pace and was much greater than the pre-training stiffness.


Escherichia coli (E. coli) and Gram-positive staphylococcus aureus (S. aureus), with 4.4 and 6.3 mm inhibition zones, respectively, being achieved by ZnPCP−10. The enhanced performance and easy degradation enables the biocomposite ZnPCP to be a prospect material in the packaging industry.Active packaging materials protect food from deterioration and extend its shelf life. In the quest to design intriguing packaging materials, biocomposite ZnO/plant polyphenols/cellulose/polyvinyl alcohol (ZnPCP) was prepared via simple hydrothermal and casting methods. The structure and morphology of the composite were fully analyzed using XRD, FTIR, SEM and XPS. The ZnO particles, plant polyphenols (PPL) and cellulose were found to be dispersed in PVA. All of these components share their unique functions with the composite’s properties. This study shows that PPL in the composite not only improves the ZnO dispersivity in PVA as a crosslinker, but also enhances the water barrier of PVA. The ZnO, PPL and cellulose work together, enabling the biocomposite to perform as a good food packaging material with only a 1% dosage of the three components in PVA. The light shielding investigation showed that ZnPCP−10 can block almost 100% of both UV and visible light. The antibacterial activities were evaluated by Gram-negative  The diversity and development of preserving and processing techniques have significantly improved food safety in recent years ,2,3,4. ABio-based materials have received increasing attention due to their environmental friendliness and structural diversity ,16,17,182− ions, which play a key role in killing bacteria and viruses [Numerous methods have been developed to improve the properties of PPL, for instance, by combining it with other functional materials such as chitosan ,30,31, a viruses . It has  viruses . However viruses . Therefo viruses . Polyvin viruses . Due to Using PPL and PVA to build the cross-linked structure along with the functional materials would endow the composite film with an active property. In our previous study, we found that ZnO prepared with the assistance of cellulose preferred growing along (001) crystal facet, and small particles were obtainable ,43. MeanThe crystalline structures of various ZnPCP samples were studied using an XRD analysis. All the samples showed the similar general patterns seen in −1 is assigned to the stretching vibration of hydroxyl group. Due to the diverse environment of hydroxyl groups appearing in PVA, cellulose and PPL, this peak was intense and broad. The C−H stretching vibration could be observed at 2910 cm−1, and the characteristic absorption peaks between 1720, 1580 and 1400 cm−1 are related to the vibrational modes of C=O and benzene skeleton. Compared to PPL, the vibrations of benzene skeleton of ZnPCP had a blue shift by 20 cm−1. This may have been caused by a hydrogen-bonding interaction between PVA and PPL, which weakens the vibrations of the benzene skeleton. The adsorption peaks around 1090 cm−1 are attributed to the vibrations of C−O in PVA, cellulose and PPL.The infrared spectra were used to characterize the functional groups, and details of the composite analysis are shown in PPL··OPVA distances were calculated to be 1.613, 1.611 and 1.758 Å for the three isomers of PPL/PVA, which have bond orders of 0.22~0.16; moreover, the related OPPL−HPPL··OPVA angles were close to 180°. The energy decomposition analysis (EDA) showed that PPL−a/PVA had a total bonding energy of −0.63 eV. The other two isomers had interfacial interaction strength in the same magnitude. In brief, PPL and PVA had interfacial chemical coupling in their composite, which was of the hydrogen-bond nature.To fully study the interaction between PPL and PVA, relativistic DFT calculations were carried out. Here, we chose three types of PPL basic units, labelled as a, b and c. Based on the finite cluster model, all possible isomeric structures were optimized. The most stable one among each type is illustrated in SEM images were taken to further explore the morphology of ZnPCP−10. From The XPS spectra of ZnPCP−10 are shown in The mechanical properties of the ZnPCP film were also tested in terms of tensile strength, elongation at break and Young’s modulus, and the results are shown in It is well known that light irradiation can affect the shelf life of food. Herein, the light shielding effect of ZnPCP films was studied using UV-visible light. Firstly, it can be seen from E. coli and S. aureus were chosen. It can be seen from E. coli and S. aureus. Among all the composite samples, the ZnPCP−10 film had the best antibacterial performance, with inhibition zones of 4.4 and 6.3 mm against E. coli and S. aureus, respectively. This good property is caused by the synergistic effect of cellulose, PPL and ZnO. First, PPL itself has many phenol groups, which can inhibit the growth of bacteria; second, ZnO can produce reactive oxygen species (ROS), which can kill bacteria [The presence of bacteria is another important factor that causes food spoilage. To test the antibacterial property of the composite films, the inhibition zone method was applied. Typical bacteria of bacteria ,53,54. SThe water content, water solubility and water vapor permeability of the composite film were also studied. The water content of the ZnPCP films was about 9.4%, which is close to that of PVA (9.86%). This indicates that introducing ZnO@PPL/cellulose into PVA has a slight effect on the water content of the film. −2 h−1 and for ZnPCP films to be between 10.8 and 7.5 g m−2 h−1. ZnPCP films exhibited a better water vapor barrier property than PVA. The low WVP would prevent water transfer between food and the surrounding environment [One of the important roles of food packaging is to reduce the water exchange between packaged food and the surrounding environment . To evalironment .Since free radicals in food are related to food deterioration and quality loss, the antioxidant property of packaging materials can also impact food shelf life . Thus, tIn order to study their fresh-keeping performance, the prepared films were used to pack fresh apple slices, as shown in 2·2H2O, NaOH, anhydrous ethanol and polyvinyl alcohol  were purchased from Tianjin Continental Chemical Reagent Factory. The cellulose was bought from Hangzhou Fuyang Beimu Pulp and Paper Co., Ltd. Agar and beef extract were purchased from Fujian Gaoxin Agar Food Co., Ltd. Peptone was purchased from Beijing Aoxin Biotechnology Co., Ltd. Larch bark was collected from Yakeshi, Inner Mongolia, China.Zn(Ac)−1.PPL was extracted from larch bark powder by an ethanol extraction method. In a typical experiment, 20 g of larch bark powder is added into 200 mL of 50% ethanol solution (1 g:10 mL) and shaken at 50 °C for 6 h. In our experiment, after filtration, brown PPL solution was obtained with a concentration of about 1.8 kg L2+ solution was obtained by dissolving 3.0 g of Zn(Ac)2.2H2O into 50 mL of distilled water. Then, 1.0 g of cellulose and 7 mL of PPL solution were added to this Zn2+ solution. The pH of the mixture was adjusted to 11 by adding 2 mol L−1 of NaOH. After stirring for 0.5 h, the mixture was transferred into a hydrothermal reactor and heated at 100 °C for 10 h. After reaction, the ZnO@PPL/cellulose was obtained by filtering, washing and drying.ZnZnPCP films were prepared by a solution casting method . First, The ZnO@PPL/cellulose dosage varied from 0.2 wt%, 0.5 wt%, 0.8 wt% to 1.0 wt%, and the resultant ZnPCP samples were named ZnPCP−2, ZnPCP−5, ZnPCP−8 and ZnPCP−10, respectively.An X-ray diffractometer  was applied to analyze the crystal structure of the samples. A Fourier transform infrared spectrometer  and scanning electron microscopy  were used to characterize the functional groups and morphologies of the samples. The chemical environments of the samples were recognized using X-ray photoelectron spectroscopy . The UV-visible transmittance of the films was measured by a TU−1950 UV-visible spectrophotometer.The ZnPCP (2 cm × 2 cm) was placed in a sealed desiccator with 50% relative humidity at 25 °C. After 48 h, sample was taken out and weighed  was dried at 105 °C for 24 h and weighed  and To test the water permeability, the samples were first treated at 23 °C and 50% humidity for 24 h to achieve a balance. After that, they were placed in a weighing bottle with 2.0 g of dried anhydrous calcium chloride. The weighing bottle was moved to a desiccator with saturated potassium sulfate solution, where the recorded temperature and relative humidity were 23 ± 1 °C and 90 ± 5%, respectively. The sample was weighed after every 2 h. WVP was determined by using the following equation:The antioxidant activity of the sample was studied using DPPH. The DPPH solution was prepared by dissolving 1.0 mg of DPPH in 20 mL of ethanol solution (the volume ratio of anhydrous ethanol to deionized water was 4:1). The 0.2 g sample was added to a 5 mL prepared DPPH solution and shaken in an oscillator for 24 h in a dark environment at room temperature . The absStaphylococcus aureus (S. aureus) and Escherichia coli (E. coli) were 1.0 × 10−4 CFU mL−1. The ZnPCP films were cut into discs with diameters of 15 mm and placed in the culture dish containing solid medium. The culture dish was then placed in an incubator at 37 °C for 24 h. Antibacterial radii were then calculated by using Equation (5):1 denotes the diameter of the outer ring of bacteriostatic sphere, and d2 is the diameter of the sample (the diameter of the inner ring of the inhibition zone).To prepare the medium, 3.0 g of beef extract, 10.0 g of peptone, 5.0 g of sodium chloride and 15.0 g of agar were added into 1000 mL of deionized water. The medium was then kept in an incubator at 37 °C for 24 h. Sterilization was performed in a portable pressure steam sterilizer at 121 °C and 0.1 kPa for 20 min. The strains of The structures of PPL and PVA and their composite isomers were fully optimized using the Priroda code . The scaE. coli and S. aureus, respectively. Due to the merits of its low cost, biodegradability and good properties, the newly synthesized ZnPCP could have tremendous potential as a packaging material.Starting from the raw materials of zinc acetate, PPL, cellulose and PVA, ZnPCP biocomposite film was successfully prepared using the hydrothermal synthesis and solution casting methods. The structural and morphological characterizations show that the growth of ZnO is tuned by cellulose, which leads to the exposure of the high-energy (001) plane. PPL covered the ZnO particles, allowing ZnO to disperse well on the PVA film. Although ZnO was coated by PPL and PVA, the coating layer was thin, as evidenced by the XPS analysis. The special structural feature could benefit ZnO, enabling it to function as an antibacterial and UV shielding agent. It was found that the best sample, ZnPCP−10, blocked almost 100% of UV and visible light. Meanwhile, it also had good antibacterial activity, with inhibition zones of 4.4 and 6.3 mm against


Researchers have investigated human altruism toward strangers for decades, using economic games such as the dictator game (DG) in their experiments. However, factors that cause the allocating behavior exhibited by those participants willing to be recipients in the DG have not been identified and the psychological mechanism of avoiding decision-making in economic games has not been widely addressed in previous studies. This study aimed to replicate previous findings regarding the number of people who are willing to be assigned the role of recipient and their allocation behavior and to explore why they share more than people who are willing to be dictators.We demonstrate that there are people willing to be assigned the role of the recipient, rather than the role of the dictator during the dictator game. In addition, we find evidence indicating that people who are willing to be recipients behave more altruistically in the dictator game than those who prefer to be dictators. Based on our results, we argue that willingness to be a recipient, in relation to the psychological unwillingness to assume responsibility and reputational concerns, is a strategic consideration.The online version contains supplementary material available at 10.1186/s13104-022-06148-3. Altruism is generally defined as behaviors that benefit another individual but are costly to the altruist ). This finding could mean that a sense of assuming responsibility increases the amount of money allocated in the DG and leads to a preference for the recipient rather than the dictator role to avoid decision-making or responsibility.Because of the small sample size, we examined all single item means and standard deviations  are willing to be recipients in the DG, and they allocate more to others than those willing to be dictators. Although the percentage of people willing to be recipients was below that in Yamagishi et al.’s  findingsPrevious studies have not comprehensively addressed the psychological mechanism of avoiding decision-making in economic games. More specifically, interpreting the reason for choosing the recipient role is not straightforward because individuals with a fairness preference would probably be willing to be assigned the dictator rather than the recipient role, as would other-regarding participants, who care not only about themselves but also others’ interests. Hence, it would be difficult to infer the existence of participants with a preference for the recipient role from the self-interested or other-regarding preferences discussed in previous studies.Exploratory analyses suggested that participants who were willing to be recipients care about the recipient’s feelings and think about their decision. These results indicate two possible strategic considerations that lead to the altruistic allocation exhibited by people willing to be recipients. One consideration is a reputational concern. Dana et al.  showed tThe other strategic consideration is about assuming responsibility. Cryder and Loewenstein  demonstrWhile the present study provides important insights that some people are altruistic and might be willing to avoid the responsibility, several limitations must be addressed. First, the sample size was small. Because of the consistency with Yamagishi et al.’s  findingsAdditional file 1. Distributions of the Amounts (JPY) Allocated to the Recipients by Participants’ Role Choice (three options)Additional file 2. Mean score and standard deviation of each item of the post-experiment questionnaire.


The rheological and pasting properties were related to the structural properties of starch, such as DI, Mw, and particle size, and were also closely related to the thermodynamic properties. The appropriate processing methods and purposes of the processed products of these yam starches can be selected according to their characteristics.In order to explore the processing and application potential of Chinese yam starch, nine kinds of Chinese yam starch  from South China were collected and characterized. The chemical composition, rheological properties, thermal properties, and in vitro starch digestion were compared, and the correlation between the structure and processing properties of these yam starches was analyzed using Pearson correlation. The results show that GY2 had the highest amylose content of 28.70%. All the yam starches were similarly elliptical, and all the yam starch gels showed pseudoplastic behavior. Yam starches showed similar pasting temperatures and resistant starch content, but SYPY showed the largest particle size (28.4 μm), SFY showed the highest setback (2712.33 cp), and LCY showed the highest peak viscosity (6145.67 cp) and breakdown (2672.33 cp). In addition, these yam starches also showed different crystal types , relative crystallinity (26.54–31.48%), the ratios of 1045/1022 cm Dioscorea opposita Thunb), the dry tuber of Dioscoreaceae plants, is an important staple food crop in many tropical and subtropical countries [Yam (ountries . In 2017Starch is the main nutrient component in yam (dry basis reaches 65.2–76.6%), and it can be used as a good source of carbohydrates in food , freeze–Yam is a traditional Chinese edible and medicinal crop. There are many high-quality local yam varieties in South China, such as Mapu yam , Lichuan yam , and Guihuai yams . In recent years, the local planting scale of these yam varieties has become larger and larger, but because there is almost no research on processing, the lack of corresponding deep-processed products has caused great waste. Furthermore, previous research to date on the function, structure, and processing application potential of yam starch has been far less in-depth than that on common commercial tuber starches such as cassava, sweet potato, and potato . This hiThe purpose of this study was to provide useful information on yam starch application in food processing by measuring and comparing the structural and physicochemical properties of nine yam starches  selected from South China. Pearson correlation was used to analyze the relationship between the structure and properties of yam starch and to further understand the influence mechanism of starch structure on its properties so as to promote the development of this agricultural industry.p < 0.05). The moisture contents of the nine kinds of yam starch were less than 11%. The highest was for MPY starch (up to 10.11%), while the lowest was for ASY starch (5.86%). As shown in the table, the amylose content of these starches ranged from 4.83% (LCY starch) to 28.70% (GY2 starch). For most of these yam starches, the content was similar to that reported for 43 kinds of yam starches (15.08–27.07%) [D. rotundata and D. alata starch (30.26% and 31.02%) [The chemical composition and molecular features of nine kinds of yam starch are shown in –27.07%)  and 10 k–27.07%)  but slig 31.02%) . This ma 31.02%) . The lip7–8.353 × 107 g/mol, 178.255–228.601 nm, and 1.381–4.959, respectively. These values are much smaller than those of the 10 yam starches reported by Rolland  [The molecular weight distribution of the nine kinds of yam starch is shown in 2–21.66) . General2–21.66) . Among tdumetorum) [The granular morphology of yam starches was observed with a scanning electron microscope. As presented in metorum) . The parmetorum) .−1 and 1022 cm−1 represent the ordered structure and amorphous structure of starch, respectively. The absorption peaks near 995 cm−1 were attributed to the bending vibration of C-OH, and the ratio of peak intensities of 1045/1022 cm−1 and 1022/995 cm−1 is an indicator of the ordered structure of starch, where the ratio of 1045/1022 cm−1 reflects the degree of order of starch. The larger the ratio, the higher the degree of order [−1 for LCY starch was 1.213, and the minimum ratio of 1045/1022 cm−1 for SYPY starch was 0.836. The maximum ratio of 1022/995 cm−1 for SFY starch was 0.712, and the minimum ratio of 1022/995 cm−1 for MPY starch was 0.568. The ratios of 1045/1022 cm−1 of the nine kinds of yam starches were larger than the ratios of 1022/995 cm−1, indicating that yam starch has a higher degree of order. This is different from the results reporting that the ratios of 1045/1022 cm−1 and 1022/995 cm−1 of seven kinds of purple sweet potato starches varied in the ranges of 0.689–0.887 and 0.850–0.974, respectively [−1 and 1022/995 cm−1 of three kinds of Chinese yam starches varied in the ranges of 0.82–0.88 and 1.10–1.14, respectively [FTIR spectroscopy has been a useful tool in monitoring structural changes in starches. of order ,10. The ectively . The ratB-type starch. LCY has strong diffraction peaks near 15°, 17°, and 23° (2θ) and weaker diffraction peaks near 5.6° (2θ), which suggests that LCY starches are C-type. At 20° (2θ), there is an amorphous peak of amylose and lipids. Yam starches  all have this peak, and their peak intensities are different. This reflects the difference between amylose content and lipid content in starches. Many studies have reported that different varieties of yam starches have pronounced diffraction peaks at 5.6° (2θ) [The crystal type of starch can be classified into A, B, and C types according to the position and number of diffraction peaks in X-ray diffraction (XRD) spectra. As shown in .6° (2θ) ,20,27. H−1 measured by FTIR. FTIR mainly measures the ordered structure in short-range molecules, while XRD reflects the ordered structure in long-range molecules [The relative crystallinity (RC) of each yam starch is shown in olecules ,29.p < 0.05). The pasting temperature of the 9 kinds of yam starch ranged from 75.37 °C to 84.37 °C, which is higher than that of the 63 kinds of yam starches reported by Muluneh (66.7 °C–78.7 °C) [w/v) [w/v) [The pasting characteristics of the nine kinds of yam starch are shown in 78.7 °C)  and the 78.7 °C) . The bre78.7 °C) . The SYP78.7 °C) . The peaC) [w/v)  and lessv) [w/v) ,34.Dynamic rheological properties can be used to determine the viscoelasticity of different samples, which is directly related to the actual processing characteristics and quality control of food. The elastic modulus (G′), viscous modulus G″, and loss tangent angle (tanδ) of the nine kinds of yam starch gels are shown in o), peak temperature (Tp), conclusion temperature (Tc), and enthalpy of gelatinization (ΔH) of the nine kinds of yam starch. The To, Tp, Tc, and ΔH of the nine kinds of yam starch were within 84.12–88.59 °C, 94.07–99.18 °C, 101.22–108.55 °C, and 23.08–31.23 J/g, respectively. In this study, the To, Tp, Tc, and ΔH values of yam starch were higher than those reported by Wang for different varieties of yam starch [p, Tc, Tc–To, and ΔH, which is related to its low amylose content. Low amylose content means that starch has higher amylopectin content, so it is more difficult to gelatinize [In the starch gelatinization process, the ordered crystalline structure is transformed into a disordered non-crystalline structure. This process is irreversible, and energy changes will occur. The differential scanning calorimetry (DSC) spectrum will show endothermic or exothermic peaks. m starch . This mam starch . LCY stalatinize . In addilatinize ,36.p < 0.05). The RS contents of the nine kinds of yam starch were significantly higher than SDS and RDS contents. The RS contents were above 90% among the nine kinds of starch; GXPY starch had the highest RS content of 97.43%, while the lowest was obtained for ASY starch (93.10%). The RS contents of 10 kinds of yam starch reported by Jiang were all above 70% [Starch digestibility is closely related to the diet and physiological function of the body, and it can have good prevention and control effects on the low starch digestibility associated with some chronic diseases. The digestibility results, including rapidly digested starch (RDS), slowly digested starch (SDS), and resistant starch (RS) of the nine kinds of yam starch, are shown in bove 70% . Among tbove 70% , RS contbove 70% . Generalbove 70% ,40,41. Tbove 70% ,43,44.p < 0.05), TV , and Tc , which indicates that the higher the amylose content, the lower the Mw, TV, and Tc of yam starch. Rg is an indirect measurement of the degree of starch branching [1  but positively correlated with PT , which indicates that the larger the Rg, the greater the limitation on the short-range order structure and the more difficult the starch gelatinization. IR1 was positively correlated with BV , Tp , and Tc-To , which indicates that the short-range order structure is an important factor affecting the gelatinization and enthalpy change of starch. DI was negatively correlated with RS , and PT was negatively correlated with RS , which indicates that the smaller the DI and PT, the greater the RS content of starch.Pearson correlation was used to analyze the relationship between the structural and physicochemical properties of the nine kinds of yam starches. As shown in ranching . Rg was p < 0.05). Mw was positively correlated with FV . Particle size was negatively correlated with FV  and G″ , which indicates that the rheological properties were related to the DI, Mw, and particle size of starch. The rheological properties and thermodynamic properties of starch were closely related. The results show that TV was positively correlated with FV , SB , and G″ . FV was positively correlated with SB  and G″ . SB was positively correlated with G″ . BV was positively correlated with Tp , Tc , and ΔH , which indicates that the G″ of starch was closely related to the viscosity change during the starch gelatinization process. The starch gelatinization process is also a process of starch enthalpy change. In this process, the starch granules expand and rupture, resulting in an increase in viscosity. Tp, Tc, and ΔH will affect the change in viscosity.The rheological properties reflect the industrial application potential of starch. The results show that DI was positively correlated with G″ , Guilin yam No. 5 (GY5), Guilin yam No. 2 (GY2), and Guangxi purple yam (GXPY) were obtained from Nanning City, Guangxi Province. Lichuang yam (LCY) was obtained from Lichuan City, Hubei Province. Shuangfeng (SFY) was obtained from Loudi City, Hunan Province. Mapu yam (MPY) was obtained from Zhangzhou City, Fujian Province. Shaoyang purple yam (SYPY) was obtained from Shaoyang City, Hunan Province. Anshun yam (ASY) was obtained from Anshun City, Guizhou Province. The species of GY11, GY2, LCY, SFY, and ASY belong to Starch was isolated using the wet-milling method previously described by Li . In detaAmylose contents, protein contents, and lipid contents of the yam starches were determined in triplicate using the method described in AOAC (1990). The moisture contents of the yam starches were determined by oven drying and heating at 105 °C until constant weight .v/v) dimethyl sulfoxide (DMSO) solution. Then, 1 mL of the suspension was removed and mixed with anhydrous methanol to precipitate the starch and then centrifuged at 4000 rpm/min for 20 min. The precipitate was re-dissolved in 5 mL of boiling water and then centrifuged at 4000 rpm/min for 30 min. The supernatant was injected into the chromatographic column , the elution rate was 0.5 mL/min, and the detectors were RI detector and small-angle light-scattering detector LALS . Based on this, the elution curve was drawn, and Origin 8.0 software was used to perform integral calculations.The molecular weight of starch was determined using a gel permeation chromatography system  . In detaThe dried starch sample was fixed on a conductive double-sided table and then sprayed with gold powder. The appearance of starch granules was observed and photographed with a scanning electron microscope . The acceleration voltage was 25 kV during micrography. The size of starch particles was scanned using a laser particle sizer  to determine the size distribution of the starch particles.−1, taking the wave number (cm−1) as the abscissa and the absorbance as the ordinate to obtain the infrared absorption spectrum.Fourier transform infrared spectroscopy of starch was performed using the KBr tablet method previously described by Shao . In detaThe crystal structure of the starch sample was measured with an X-ray diffractometer  following the method described by Jiang . In detaPasting properties of yam starches were determined using the method previously described by Jayakody . In detaDynamic rheological properties of yam starches were determined using the method previously described by Zhu . In detaw/w) suspension of starch in water was prepared, sealed in a DSC crucible, and then kept at 25 ± 2 °C for 4 h to ensure that the starch samples were fully mixed with water. The empty crucible was used as a control. The samples were scanned from 20 °C to 200 °C at a rate of 10 °C/min under a nitrogen flow rate of 150 mL/min. The gelatinization enthalpy (∆H) and the onset (To), peak (Tp), and conclusion (Tc) gelatinization temperatures of each sample were then determined from thermograms. All analyses were performed in duplicate, and mean values were calculated.Thermal properties of yam starches were determined using a differential Scanning Calorimeter  following the method previously described by Zhang . In deta20 and G120 were calculated. The in vitro digestibility of starch was evaluated by three indicators: rapidly digested starch (RDS), slowly digested starch (SDS), and resistant starch (RS) . The formulas are as follows:In vitro starch digestion of yam starches was determined using the DNS method with reference to Amruthmahal . In deta20 (mg) is the glucose released after 20 min of enzymatic hydrolysis. G120 (mg) is the glucose released after 120 min of enzymatic hydrolysis. FG (mg) is free glucose. TG (mg) is total glucose. TS (mg) is the dry basis weight of total starch, and 0.9 is the conversion factor.Remark: Gp < 0.05, and all data are expressed as mean ± standard deviation (SD).Excel 2010, Origin 8.0, and SPSS 20.0 were used for data processing and analysis, and the Duncan test method in the analysis of variance (ANOVA) was used for the significance analysis. The significance level of the test was In this study, the structural and processing properties of nine kinds of yam starch were compared, and the correlation between the structure and properties of yam starch was explored. The results show that GY2 had the highest amylose content of 28.70%. All yam starches showed similar morphology , and all the yam starch gels showed pseudoplastic behavior. However, these yam starches showed different crystal types . GY2 showed the highest relative crystallinity (31.48%), SFY showed the highest setback (2712.33 cp), and LCY showed the highest peak viscosity (6145.67 cp) and breakdown (2672.33 cp), but all yam starches showed similar pasting temperature and resistant starch content. The particle size and molecular structure of different varieties of yam starch were also different. From the results of Pearson correlation analysis, the higher the amylose content, the lower the Mw, TV, and Tc of yam starch. The larger the Rg, the greater the limitation on the short-range order structure and the greater the impact on starch gelatinization and enthalpy change. The rheological properties were related to the structural properties of starch, such as DI, Mw, and particle size, and were also closely related to the thermodynamic properties. Appropriate processing methods and products can be selected according to the different characteristics of different varieties of yam starch. This study provides useful information for the application of these yam starches in food.


We investigated whether (1) imported pre-cut tissue is feasible for Descemet membrane endothelial keratoplasty (DMEK) in eyes of Asian patients, (2) the clinical outcome is comparable between the endothelium‐in and endothelium‐out methods, and (3) the corneal edema-induced anterior curvature changes may have an effect on the refractive error.The medical records of 32 DMEK patients who underwent either the endothelium-out or endothelium-in method using imported pre-cut grafts with a 3-day pre-cut-to-use time were retrospectively analyzed. Fuchs’ endothelial dystrophy (37.5%) and bullous keratopathy (62.5%) cases were included. The main clinical outcome measures were graft survival, best corrected visual acuity (BCVA), endothelial cell density (ECD), corneal thickness (CT), and complications. Correlation of the anterior curvature changes with refractive error was analyzed in the DMEK with cataract surgery group.2, respectively. The mean survival time, ECD, BCVA, CT, and complication rates were not different between the endothelium-in and endothelium-out methods. A hyperopic shift by +0.42 D was not related to the anterior curvature changes.The overall survival rate was 71.9%. Final graft failures were caused by rejection, glaucoma, and infection. Visual acuities improved by 89.3%. BCVA better than 20/40 and 20/20 was found in 75% and 28.6% of patients, respectively. The ECDs at 3 months and 1 year were 1400 and 1083 cells/mmImported pre-cut tissues with a ≤ 3-day pre-cut-to-use time are feasible for DMEK in the treatment of corneal endothelial edema in eyes of Asian patients, and both endothelium-in and endothelium-out methods appear to be comparatively effective. Edema-induced anterior curvature change may not affect the refractive shift. Since Gerrit Melles first proposed the concept of posterior lamellar keratoplasty in 1998 , 3. AfteSince various surgical techniques are employed in DMEK to reduce the endothelial cell loss , 14–19, Additionally, the refractive outcome and factors causing shift in the refractive errors have been currently debated in patients with FED following DMEK combined with cataract surgery –23. VariOverall, three issues need to be addressed: (1) whether imported pre-cut tissues are feasible for use in DMEK in Asia, (2) whether the clinical efficacy of the endothelium‐in delivery method is comparable to that of the endothelium‐out method in DMEK, and (3) the extent of shift in the refractive error following DMEK and whether it is related to the anterior curvature changes. Therefore, this study investigated 1) the clinical efficacy of DMEK in eyes of Asian patients using imported pre-cut tissues with a ≤ 3-day pre-cut-to-use time, 2) the correlation of the anterior curvature changes with refractive shift in DMEK combined with cataract surgery, and 3) whether the clinical efficacy of the endothelium‐in delivery is comparable to that of the endothelium‐out method in DMEK with imported pre-cut tissues. These results will benefit the standardization of DMEK procedures in eyes of Asian patients using imported pre-cut tissues.th in 2022 were retrospectively reviewed.The study adhered to the tenets of the Declaration of Helsinki. Informed consent was waived by the Institutional Review Board because the study was based on the retrospective review of old charts. Under the approval by the Institutional Review Board of Seoul National University Hospital (IRB No. 2108-026-1241), the medical records of DMEK cases performed by one surgeon (M.K.K) at our tertiary referral center between October in 2016 and May 6Patients who had been diagnosed with corneal endothelial edema due to either FED or BK and had undergone DMEK with ≥6 month-follow up were included. The surgical indication was persistent corneal stromal edema with decreased visual acuity. Among the 35 DMEK cases in the above-mentioned period, three cases were excluded from the analysis. Two were excluded owing to loss in follow-up (< 6 months), and the remaining was excluded because the patient had previously undergone whole corneal transplantation prior to DMEK with a high risk of rejection. Of the total 32 cases, one-third had grade 4 FED with corneal stromal edema , and two-third had BK  , 25. EyeThe following data were collected from the medical charts: demographic information that included the recipient’s age, sex, general medical history, ocular medical and surgical history; and ophthalmic findings that included visual acuity, intraocular pressure (IOP), endothelial cell density (ECD), central corneal thickness (CCT), ocular biometric parameters, and the follow-up period . Concomiin situ on top of the donor corneal stroma in Optisol . The mean pre-cut-to-use period for overseas transport was 59±15 hours. Donor tissue information, including age, graft size, donor ECD, and death-to-use period is shown in A total of 32 transplant grafts were supplied by the Eversight Eye Bank  using the standard procedure , 27. OnlPeripheral laser iridotomy or surgical iridectomy during cataract surgery was performed before DMEK. For patients with a cataract or a pupil anomaly, phacoemulsification with an intraocular lens (IOL) insertion or pupilloplasty was performed concomitantly or sequentially prior to DMEK without any adverse event. Endothelium-out grafts were delivered into the anterior chamber during DMEK using glass Jones tubes  in 17 eyes from October 2016 to March 2019. After August 2019, a Coronet EndoGlide insertion device  became available in Korea; after August 2019, 15 eyes received endothelium-in grafts. The size of the recipient’s Descemet membrane stripping was usually 0.5–0.75 mm larger than that of the donor graft (8.0–9.0 mm).In the endothelium-out technique, the graft was freed from the hinge and spontaneously rolled with the endothelium outward. Using a Jones tube, the graft was aspirated inside the tube with a balanced salt solution  and injected into the anterior chamber through a 3.5-mm corneal or scleral tunnel. The graft was unfolded using tapping procedures and was attached using an air tamponade (80–95%) after confirming that the endothelium was positioned downward by the S mark  28]..28].In the endothelium-in technique, the graft was harvested and tri-folded inward with its endothelium. The graft was pulled into the coronary EndoGlide, which had been filled with the balanced salt solution. The graft was delivered into the anterior chamber in the correct orientation with the endothelium facing downward through a 2.75-mm corneal or scleral tunnel. The graft was unfolded with the assistance of tapping and was attached using an air tamponade (80–95%) with a confirmation of the S mark  14]. Th. Th14]. All the patients received systemic steroid treatment for more than 3 weeks. Topical 1% prednisolone acetate  was administered 4 times a day and the dose was tapered down to 1/day after 12 months. Topical 0.5% moxifloxacin eye drops  were applied 4 times a day.postoperatively measured) [pre-post (CCTpreoperative−CCTpostoperative) or ΔKpre-post (Kpreoperative−Kpostoperative) with MNE was analyzed. In most patients, the preoperative posterior curvature (Kpost) data were not available. Therefore, postoperative Kpost (n = 15) was compared with the Kpost in eyes of normal controls (n = 50) in whom the parameters for cataract surgery were measured using an IOL Master 700 .The following data were collected as the main outcomes and analyzed in each time point in a whole group and were also compared in between the endothelium‐in and endothelium‐out deliveries; graft survival, postoperative best corrected visual acuity (BCVA), ECD, CCT, re-bubbling rate and annual EC loss (ECL) rate. Early and delayed complications were additionally compared between those two groups. Visual acuities measured initially by Snellen charts were converted to logarithm of the minimum angle of resolution (logMAR) values for analysis. The proportions of BCVA ≥20/40, 20/25, or 20/20 were evaluated. Patients with amblyopia, advanced glaucoma, or macular disease that could affect visual acuity were excluded from visual acuity analysis. Increased IOP was defined as IOP > 21 mmHg. ECD was checked by specular microscopy . CCT was measured with anterior segment optical coherence tomography. The postoperative immediate and delayed complications were examined by slit-lamp microscopy . To analyze the changes of corneal astigmatism, axial length (AXL), and mean numerical refractive errors (MNEs) in DMEK combined with cataract surgery (n = 18), an auto-kerato-refractometer , topography , and IOL master  were used. MNEs were calculated by subtracting the value of postoperatively measured refractive error from the value of the predicted refractive error calculated by IOL master .Data are presented as mean ± standard deviation where applicable. Statistical analysis was performed using SPSS Statistics 20.0 . The chi-squared test or Fisher’s exact test was used to compare the two categorical variables, and Student’s t-test was applied to compare the means of continuous variables between two groups. The Kaplan-Meier curves were used to analyze the corneal graft survival after a successful DMEK and a statistical difference in Kaplan-Meier curves was assessed by Log-rank test. Pearson’s bivariate co-relation was used between those ocular biometric parameters. A P value of < 0.05 was considered statistically significant.The baseline characteristics of the donors and recipients are shown in Among the donors’ clinical features, the graft size was significantly larger in the endothelium-in group (8.0±0.4 mm) than in the endothelium-out group . The mean donor death-to-use period in the endothelium-in group (135±8 hours) was similar to that in the endothelium-out group . The mean pre-cut-to-use period did not differ between the two groups . There was no difference in the ECD between the two groups.2 (n = 8). After DMEK, the mean ECD was 1400±480 cells/mm2 at 3 months, 1291±497 cells/mm2 at 6 months, 1086±384 cells/mm2 at 1 year, and 951±252 cells/mm2 at 2 years , and +0.40±1.5 D and +0.45±1.4 D in the endothelium-out and endothelium-in groups, respectively . Postope p>0.05; . MNE als p>0.05; . Mean vaThis study demonstrates that 1) the clinical efficacy and safety of DMEK in eyes of Asian patients using imported pre-cut tissues with a < 3-day pre-cut-to-use time are comparable to those in penetrating keratoplasty for BK, 2) the efficacy of the DMEK endothelium‐in method is comparable to that of the endothelium‐out method, and 3) a hyperopic shift by +0.42 D and a significant reduction of corneal astigmatism were observed. Corneal edema-induced anterior curvature changes may not affect the refractive error. However, the ECL within a month and the early annual rates of ECL were greater than those previously reported , 29–31.Since endothelial keratoplasty, including DMEK, DSAEK, and ultrathin-DSAEK, results in better visual acuity and lower graft rejection than those by penetrating keratoplasty , 3, 7, eInitial ECL at 6 months varies from 21 to 56% in reports, based on the proportion of the included FEDs and the various techniques of graft delivery to the anterior chamber , 36, 37.st year; 16.5±11% vs 17.8±12% at 2nd year, p > 0.05). A high proportion of BK inclusions (62.5%) also contributed to the high ECL at the early stage. However, similar to a DSAEK study using imported corneas [Advanced FED with stromal edema was included in this study, whereas FED eyes without significant corneal edema were included in reports from Western countries , 25, 34. corneas , this st corneas , the surin vitro studies discovered that ECL with the tri-folded endothelium-in method was comparable to that with the scroll-based endothelium-out method [Currently, graft delivery and unfolding procedures need to be standardized to improve the surgical outcomes. Critical steps are still performed in variable ways among different surgeons. To minimize either delivery-associated or tapping-induced ECL, we compared the surgical outcomes between the endothelium-in and endothelium-out deliveries. The visual acuity, ECL, and CCT did not differ between the two methods. Recently, t method –42. Moret method . Since tt method , 43, 44.t method Since the dark pigmented iris blocks visibility to the graft, graft positioning with an appropriate orientation is difficult in eyes of Asian patients. Therefore, the tri-folded endothelium-in method with the assistance of graft-grabbing forceps may be an easier procedure with fewer upside-down placements than the endotheium-out method. In most cases, the scleral tunnel is preferred because of the small dimensions of the anterior chamber. Since the Geuder glass cannula is not currently available in Korea, a modified Jones tube with a 3.5 mm incision or an EndoGlide with a 2.75 mm incision are possible options. Using EndoGlide could be beneficial in maintaining anterior chamber stability during insertion compared to the Jones tube.There were no differences in postoperative complications between the endothelium-in and endothelium-out delivery. One patient developed a pupillary block following re-bubble owing to discharge and early ambulation of the patient. Inferior iridectomy is usually recommended for preventing air- or gas-induced pupillary blocks –47. Howepreoperative and ΔKpre-post indicates anterior curvature changes between before and after DMEK, changes of neither CCT nor anterior curvature correlated with the refractive shift, which supports the study by Augustine et al [post) between the post-DMEK and normal control eyes, suggesting that the graft does not induce a major change. However, we could not evaluate directly whether the individual changes of the posterior curvature could affect the refractive shift. Considering that previous studies show that the posterior cornea curvature correlates with the hyperopic shift of post-DMEK eyes [This study demonstrated a hyperopic shift (+0.42 D) which corresponded with the refractive outcomes (+0.55 ~ +1.12 Diopter) of previous studies following triple DMEK , 52, 53.ne et al . AnotherMEK eyes , 23, 54,post was not measured since the preoperative data were not available. Therefore, further studies to assess the correlation of the Δ Kpost with MNE are warranted. Lastly, DMEK was performed by a professor (MK Kim) who had previous experience of performing approximately 1000 penetrating keratoplasties and 30 deep anterior lamellar keratoplasties. Before starting DMEK, the professor had undergone hands-on training programs by a DMEK wet lab course conducted by the Gerrit Melles of Netherlands Institute for Innovative Ocular Surgery (NIIOS) Academy in Rotterdam. This study reported an incidence of learning curve related-early graft failure of 6.3%, which was comparable to the previous incidences of learning curve related-early graft failure of 7.69 to 4% [This study has several limitations. Owing to the limitations of its retrospective design, some of the data were missing. The small sample size owing to the low popularity of DMEK surgery could have affected the statistical power. However, considering that other studies comparing two groups with similar sample sizes , 42, 55,69 to 4% –62. AlthIn conclusion, performing DMEK using imported pre-cut tissue with a ≤ 3-day pre-cut-to-use period may be a feasible option for either BK or FED in eyes of Asian patients. Additionally, the tri-folded endothelium-in method seems to be comparatively effective to the endothelium-out method for DMEK with imported pre-cut tissue. A hyperopic shift may not be related to the anterior curvature changes.S1 FigData are expressed as mean ± standard error of the mean.(TIF)Click here for additional data file.


Liver transplantation (LT) has become a victim of its own success. Advances in the field of LT have led to superior outcomes, and patients with diseases such as hilar cholangiocarcinoma and colorectal liver metastases, which were once considered as contraindications for surgery, are now eligible for transplantation. This expanded eligibility has forced LT practitioners to increase the size of the donor pool by expanding the acceptance criteria  and investigating alternative solutions to rescue organs that would otherwise be discarded because they carry a high risk of post-transplant failure.Attempts to widen the pool of donors in order to meet demand has yielded tissue from older, fatty livers and comorbid donors and led to donation after circulatory death (DCD), in addition to tissue retrieved from traditional dead-brainstem donors (DBD). Grafts from these extended criteria donors (ECD) have higher rates of primary nonfunction, early graft dysfunction, and the formation of non-anastomotic biliary strictures when transplanted. These postsurgical side effects worsen patient outcomes and increase healthcare costs. Additionally, transplant surgeons are less likely to use ECD grafts after inspecting the quality of the tissue.Given the current trends in aging, obesity and diabetes, it is expected that the annual utilisation rate of liver grafts in the US will fall from a peak of 78% in 2010 to 44% by 2030, thus decreasing the number of LTs performed [The concept of MP is not new. Thomas Starzl and his team used the technique upon the inception of LT procedures. Brettschnider first described the outcomes of experiments using MP for canine livers. The researchers performed dual perfusion of the portal vein and hepatic artery with heating and hyperbaric oxygenation and compared these tissues to cold perfused livers. While the cold perfused livers required immediate transplantation, they found that the livers treated with MP could be sustained for 8 to 24 h before transplantation [While SCS reduces the graft metabolism by 10–12 times, the anaerobic activity continues, leading to the generation of oxygen free radicals and, ultimately, ischaemia-reperfusion injury when the clamps are removed. The basic principle of MP is that cellular respiration can replace cold ischaemia. In practice, there are several methods used to achieve this. Two common methods are hypothermic MP (HMP) and normothermic MP (NMP).In HMP, the cellular metabolism and ATP demand are low. The HMP grafts are connected to a pump that supplies the portal vein with preservation fluid, similar to the Belzer solution—a commonly used cold perfusion fluid. The mechanism of action is considered to work by removing metabolic waste products and protecting the endothelium. Building on this technique, oxygen can be added to the portal perfusion circuit in order to perform hypothermic oxygenated machine perfusion (HOPE). Additionally, oxygen may be added to the hepatic arterial circulation in order to provide a dual oxygenated circuit (D-HOPE), in which the portal vein and hepatic artery are perfused with oxygenated fluid. Experimental results have shown that using oxygenated preservation fluid reduces the accumulation of succinate in the mitochondria, which reduces the production of oxygen free radicals and the likelihood of ischaemia-reperfusion injury.A randomised, multicentre trial investigating D-HOPE vs. SCS after DCD demonstrated a decreased incidence of intraoperative postreperfusion syndrome , graft dysfunction, and biliary complications in patients who received grafts perfused using the D-HOPE protocol. Furthermore, four times fewer post-transplant biliary interventions were reported in the D-HOPE group compared to the SCS group .While the previously mentioned trial only assessed DCD livers, HMP can also be used for ECD-DBD livers. A multicentre, randomised trial investigating HOPE vs. SCS in the latter group of donors reported a reduction in the postoperative peak ALT levels, complications at the 90-day postoperative mark, and the length of hospital stay for patients who received livers treated with HOPE. In contrast to the trial examining the effects after DCD, the ECD-DBD recipients did not have significantly lower rates of graft dysfunction or biliary complications . In summThere are a few differences between HMP and NMP. As the hepatocyte metabolism in the graft is suppressed during MP, there are two consequences. First, HMP has a smaller theoretical risk of graft loss than NMP. Additionally, if there is a failure in MP and the perfusion ceases, a graft connected to HMP will not be subjected to warm ischaemia. Furthermore, keeping the graft in hypothermic conditions allows for conventional retrieval, initial flush, and transport in SCS and does not require the transportation of specialised equipment by the retrieval team.In contrast, NMP preserves the liver in near-physiological conditions. The system uses a blood-based perfusate that maintains the cellular metabolism and experimentally inhibits inflammation and prevents cell death. Previous experiments assessing gene expression in NMP grafts identified significant activity in terms of growth, metabolism, and tissue repair . The perIn one retrospective study, 31 ‘orphan’ ECD grafts, which were refused by all LT centres in a UNOS region, were accepted and placed on HMP after SCS. Viability testing was conducted by sampling the HMP effluent prior to transplantation. Comparing the patient outcomes with a historical matched cohort led to the identification of improved outcomes for HMP patients in terms of early graft dysfunction, the hospital length of stay, and biliary complications at 1-year post-transplantation. This trial demonstrated that ECD grafts can be safely tested and transplanted, that HMP can overcome the logistical problems of transferring organs between regions, and that the grafts can be tested for viability .The situation of NMP is more complicated than that of HMP because there are inconsistencies in the application of NMP. Some protocols minimise the cold ischaemia time by initiating perfusion with a transportable MP at the retrieval hospital, and others initiate MP at the transplant centre after the SCS and transport. Both methods have been associated with the increased use of ECD grafts and superior outcomes compared to SCS ,8.One multicentre, randomised trial compared NMP to SCS in adult DBD and DCD patients. Despite identifying longer functional warm ischaemia and overall storage times in the NMP DCD grafts, the authors reported better outcomes with the use of NMP. Indeed, NMP DCD grafts had better rates of early allograft dysfunction and peak AST levels compared to the SCS DBD grafts. However, the rates of biliary complications in this study were not significantly different, as both DBD and DCD grafts were included .While these studies demonstrated the utility of MP, they raise questions as to which type of MP is most effective, given the circumstances of the donor, the recipient, and available logistics. Another conclusion that can be drawn is that, compared to SCS, MP has shown no benefits for patient or graft survival in any of the trials mentioned above. This might be because primary nonfunction and mortality are relatively uncommon events compared to early graft dysfunction.A less widely published technique of MP is to perfuse the organs before they are removed from the donor. Normothermic regional perfusion (NRP) involves placing cannulae in situ, before or during tissue retrieval. Normothermic perfusion seems to be the ideal technique for collecting tissue from DCD. The withdrawing of life support in this group is associated with physiological changes in peripheral and mesenteric vasoconstriction caused by catecholamine release upon brainstem injury. This occurs before the declaration of death and is compounded by warm ischaemia prior to cross-clamping. Thus, even before circulatory arrest and the declaration of death, grafts can become ischaemic, which cannot be stopped by cold perfusion. In NRP, oxygenated blood perfuses the abdominal organs via an extracorporeal circuit consisting of an oxygenator, heater, and pump. The circuit is completed soon after the declaration of death and continues running for 2 h, after which a cold perfusion fluid is run through the circuit. A randomised trial found significantly lower rates of early graft dysfunction (12% vs. 32%), no instances of cholangiopathy (0% vs. 27%), and a doubling of the organ utilisation rate (61% vs. 27–36%) for NRP compared to standard DCD, respectively. In France, Italy, and Norway, abdominal NRP is the standard procurement procedure for DCD . It is npost-mortem cannulae placement [However, using NRP raises the ethical issue of when the cannulae should be inserted into the donors. While the premortem placement of endovascular occlusive balloons and perfusion cannulae is beneficial for the grafts, initiating the retrieval process before death might be unacceptable for some patient groups. However, in the study by Watson et al., the reported benefits were observed following the lacement .A common variable in all these studies is the effect that MP has on logistics. In most cases, the effect is beneficial, as time on MP often means that LT can be delayed until daytime, which is generally considered a safer time to operate. In other ways, MP is cumbersome, as perfusion devices and technicians are required to monitor the equipment and may be required at the retrieval hospital. This requirement increases upfront costs; however, this should be mitigated by downstream cost savings from reduced morbidity. Finally, MP has been shown to increase the utilisation of grafts in ECD. Thus, the appeal of MP has come full circle, to the point when Brettscheider first demonstrated its efficacy.In conclusion, several systems exist for performing MP using LT grafts. Although the techniques vary in terms of their mechanisms and applications, thus far, all techniques have shown benefits for LTs from ECD-DBD and DCD donors. Determining which method is appropriate in specific circumstances is the next challenge for researchers. Each MP system has benefits, and it might be the case that HMP and NMP are complementary techniques. The results of upcoming multicentre trials, expected in the next 2 years, should elucidate these differences.