Little is known about the connection between preoperative keratometry and postoperative results of myopic small-incision lenticule extraction (SMILE). To determine the influence of extreme (flat and steep) corneal keratometry on the safety and efficacy of SMILE, the databases of the Department of Ophthalmology, Ludwig-Maximilians-University Munich, Germany, and SMILE Eyes Linz, Austria, were screened for patients with steep and flat keratometry who had undergone SMILE. In this cross-sectional matched comparative cohort study, eyes with markedly flat  or steep (≥ 47.0D) preoperative corneal keratometry were matched to a cohort of eyes with regular keratometry (42.0–46.9D) by preoperative manifest refractive spherical equivalent and cylinder, age, corrected distance visual acuity and surgical SMILE parameters. The standardized graphs and terms for refractive surgery results were applied to compare the three groups. Changes in higher order aberrations (HOAs) were evaluated on Scheimpflug imaging. In total, 63 eyes (21 each) of 54 patients with a mean refractive spherical equivalent of  − 5.21 ± 1.59 D were followed up for a mean of 9.2 ± 6.1 (minimum ≥ 3) months. Mean baseline keratometry was 41.3 ± 0.7D (flat), 45.5 ± 1.0D (regular) and 47.7 ± 0.6D (steep) (p < 0.0001). Compared to the regular group, the flat and the steep cornea group resulted in a non-inferior percentage of eyes within ± 0.50 D of target refraction (p = 0.20), uncorrected distance visual acuity (p = 0.95) and corrected distance visual acuity (p = 0.20). Flat corneas however experienced a stronger induction of spherical aberration (SA) compared to the steep group (p = 0.0005). In conclusion, non-inferior outcomes of SMILE can also be expected in eyes with steep (≥ 47D) or flat (< 42D) preoperative keratometry, while SMILE however induces more SA in eyes with a flat keratometry. The former offers on-par results concerning safety and efficacy13. SMILE performed with the VisuMax platform  requires direct contact with a disposable interface of variable diameter, but uniform radius, which represents a “one-curvature-fits-all” approach. Therefore, in steep corneas, excessive applanation  might cause tissue irregularities within the treated stromal tissue. Moreover, in flat corneas, excess suction may be required to accommodate the corneal surface into the relatively steeper contact glass , preoperative keratometry can influence postoperative refractive and visual outcomes, especially in extremely flat or steep keratometry, and even more so in conjunction with high myopic ablations13, data on SMILE is still lacking. Hence, the objective for this study was to compare the outcomes of myopic SMILE in eyes with excessively steep and flat preoperative keratometry to regular keratometry regarding safety, efficacy and higher order aberrometric outcome.While there is plenty of literature on the relationship between preoperative keratometry and the outcomes of LASIK and photorefractive keratectomy (PRK)For the purpose of this cross-sectional study, the database of the SMILE Eyes Clinic  encompassing more than 1800 SMILE procedures was screened for eyes with preoperative flat (< 42D) or steep (≥ 47D) keratometry using the K1 (steeper meridian) value. Within these two groups, eyes were matched concerning preoperative manifest refraction spherical equivalent (MRSE), manifest sphere and cylinder, age, preoperative corrected and uncorrected distance visual acuity  and surgical SMILE parameters . Eyes that did not find a matching partner were removed from analysis. Afterwards, secondary matching to a third group with regular keratometry (42.0–46.9D) was performed based on the same parameters mentioned above. Minimum follow-up was 3 months. Informed consent from each patient as well as Institutional Review Board approval from the Department of Ophthalmology, Ludwig Maximilian’s University was obtained prior to the surgical procedure and data analysis. All study procedures adhered to the tenets of the Declaration of Helsinki and relevant local guidelines and regulations.At first presentation, a standardized preoperative protocol was followed as established for all patients undergoing corneal refractive surgery. This included slit lamp-biomicroscopy and fundoscopy, objective, subjective and cycloplegic manifest refraction and Pentacam corneal tomography  to rule out contraindications for keratorefractive treatments (e.g. keratoconus).16 with a standard cap thickness of 120 to 140 µm and an optical zone of 6.4 to 6.7 mm. In brief, surgery was performed under topical anesthesia with oxybuprocaine hydrochloride  using a 500 kHz femtosecond laser . After sterile draping, the eye of the patient was placed under the femtosecond laser’s operating microscope, centered and immobilized with a suction contact glass. The size “small” was used in all cases. All patients were operated by one of two experienced surgeons  using the same laser scanning settings (4.5 µm spot spacing for the cap and lenticule interface at 160 nJ laser energy).SMILE was performed as described previouslyOne side cut of 4.00 mm arcuate length was created at the 135° and 45° position in the right and left eyes, respectively. After manual dissection using a blunt spatula, the lenticule was removed through the incision as described previously and checked on the corneal surface for completeness. Then, the interface was irrigated with balanced salt solution. The post-operative treatment regimen consisted of tobramycin/dexamethasone combination eyedrops  six times daily for the first week, dexamethasone  eyedrops from the second until the fourth week four times daily, and lubricant eye drops tapered as needed.17. Statistical analysis was performed in SPSS Statistics 23 . Normality of data was assessed with the Kolmogorov–Smirnov test. Homogeneity of variance was checked using Levene’s test. Analysis of variance (ANOVA), the Kruskal–Wallis and the Chi-Square test were used to compare the groups as appropriate. The independent samples t-test and Mann–Whitney U test were performed as post-hoc tests with Bonferroni-Holm correction to compensate for multiple testing. Comparing all three groups, the level to indicate statistical significance was defined as p < 0.05; using Bonferroni’s correction, the level of significance was adjusted to p < 0.0167 for post-hoc analyses.All data were gathered and analyzed in Microsoft Excel spreadsheets  according to the established graphic reporting of outcomes of refractive surgeryA total of 63 eyes 54 patients were included in the study, with 21 eyes in each of the flat, regular and steep group. Mean preoperative MRSE was − 5.21 ± 1.59 D and surgical target refraction was plano in all eyes, aside from 2 eyes (9.5%) in both the flat and steep group and 4 eyes (19.0%) in the regular group that were targeted for mini-monovision of up to − 1.0D. The mean postoperative follow-up was 9.2 ± 6.1 months. Patients’ baseline parameters can be found in Table There were no intra-, peri- or postoperative complications in either group. There was no loss of two or more lines in any group Fig. . Loss ofThere was no difference in refractive accuracy or stability at the end of follow-up . The percentage of eyes achieving 20/20 and better UDVA was comparable between all groups . The efficacy index was similar across all groups .Mean K values decreased from 41.3 ± 0.7D (flat), 45.4 ± 1.0D (regular) and 47.7 ± 0.6D (steep) to 36.8 ± 1.3D, 40.6 ± 1.6D and 42.9 ± 1.0D, respectively . The ratio between the surgically induced change in MRSE and the surgically induced change in mean K values was 1.22 (flat) vs. 1.20 (regular) vs. 1.23 (steep) with omnibus p = 0.92. The corresponding Spearman correlation coefficients (Rho) were 0.53 (p < 0.001), 0.59 (p < 0.001) and 0.65 (p < 0.001), respectively.Preoperatively, the flat group showed significantly lower values of total HOAs than the two other groups , coma (p = 0.30) and trefoil (p = 0.10) were induced by SMILE to a comparable extent in all groups. The flat group, however, experienced a significantly higher induction of spherical aberration than the steep group , and a tendentially higher induction of spherical aberration than the regular group .The present study was designed to test the safety, efficacy and higher order aberrometric outcome obtained by myopic SMILE in eyes with extremely flat and steep keratometry readings. Analyzing the outcomes in three matched groups, we concluded that SMILE produced non-inferior results in flat (< 42 D) and steep (≥ 47 D) corneas as compared to the regular control group matched by age, follow-up, MRSE, sphere, astigmatism, CDVA, UDVA and SMILE parameters.forme fruste of keratoconus. Since values between 47.2 and 48.7 D might be regarded as “probable”, and values > 48.7 D as “probably definite” keratokonus18, refractive surgery in such eyes requires thorough bilateral ruling-out of pre-existing ectatic disease, e.g. using the Belin-Ambrósio enhanced ectasia screening based on Scheimpflug imaging19. Although most case reports published describe ectasia after 6 to 12 months after SMILE, a time frame encompassed in our study, long-term follow-up remains warranted in such steep keratometry after keratorefractive treatment. Conversely, the majority of patients reported with ectasia after SMILE already had suspicious topography and keratometry preoperatively20, which was carefully ruled out in every eye included in this study.Concerning safety, all groups exhibited an excellent profile. No eye lost two or more lines of CDVA, and only 10% of eyes in the regular group lost one line. Moreover, no ectasia was observed during the mean follow-up of 9.2 months, bearing in mind that eyes with steep keratometry might predispose to, or display a sub-clinical Concerning efficacy, both groups with flat and steep corneas showed similar, comparable results in the final MRSE and refractive accuracy as compared to regular keratometry. However, the percentage of patients with postoperative manifest cylinder of < 0.50 D was lower, and the total value of astigmatism was significantly higher in the steep than the flat group. Nevertheless, these subtle, clinically probably irrelevant differences did not translate into differences in visual acuity. In our study, both the flat and steep groups achieved comparable, non-inferior percentages of eyes achieving UDVA of 20/20 or better as compared to the regular group. Although it is unclear whether a larger sample size would have been able to determine, even if subtle, inferiority in astigmatic target accuracy in extremely steep corneal curvature, the non-inferior visual results reported in our study might suggest that any detectable differences might be more of academic, and less of clinical value.In addition to the standard reporting of refractive outcomes, our study performed an in-depth analysis of HOAs and their relationship to preoperative corneal curvature. Theoretically, discrepancies between the uniform radius of the SMILE contact glass interface and extreme corneal keratometry might result in tissue compression or suction, thus distorting the stroma. This could entail the creation of irregular cap or lenticule interfaces with potential ramifications on iatrogenic HOA induction. Interestingly, neither the flat nor the steep group showed increased HOA values as compared to the regular group, corroborating this hypothesis. Of note, we found that eyes in the flat keratometry group responded to myopic SMILE with significantly more spherical aberration induction than the steep, and tendentially more than the regular group. As stated above, this might result from the forced accommodation of the flat corneal surface into the relatively steeper contact glass, causing irregular lenticule borders within the stroma after tissue relaxation. The clinical consequence of this finding may be negligible as the flat keratometry group exhibited the lowest preoperative level of spherical aberration of all three groups, that was still lowest of all groups postoperatively.Limitations of this pilot study mainly include its small sample size. Larger studies are warranted to exclude differences not detected in this study with only 21 eyes per group; and moreover, such studies should also be designed with a homogenous structured follow-up with twelve-month data, which our pilot study could not provide as yet .In conclusion, this matched comparative study suggests that myopic SMILE also produces safe and effective, non-inferior refractive and visual outcomes in eyes with extremely flat and steep keratometry. In eyes with excessively steep keratometry, utmost caution should be taken to exclude underlying ectatic disease.


In addition to deficits in pragmatics, children with autism spectrum disorders (ASD) have weaknesses in complex syntax and working memory (WM). These two deficits may be closely related. Previous work investigated the effects of WM training in developmental language disorders and showed significant improvement in both WM and syntax. The current study tests the impact of 12 h of WM training across 8 weeks in 30 children with ASD, aged 5 to 11. Results showed direct improvements on untrained WM tasks, as well as transfer effects to syntax and processing speed. Stronger WM led to better syntactic abilities. While they must be replicated, these exciting results provide impetus for further studies of WM interventions.The online version contains supplementary material available at 10.1007/s10803-021-05295-z. Autism spectrum disorder (ASD) is a neurodevelopmental condition characterized by persistent deficits in social communication and social interaction, as well as restricted and repetitive patterns of behavior, interests and activities  confirmed deficits in both verbal and visuospatial components of WM, where neither age nor IQ explained the observed WM differences. Another meta-analysis published by Wang et al. in  is also linked to syntax. For example, Adams and Gathercole  and WillDelage and Frauenfelder  conductele lave ‘he’s washing him/it’), Durrleman and Delage (me lave ‘he is washing me’), which do not require the morphological marking of gender and number,Numerous studies have examined the link between WM and syntax in children with atypical development. Stanford  reportedd Delage  found a Delage and Frauenfelder  evaluateMagic Memory  complex syntax abilities, WM training in this population was the logical continuation of this research area. Our previous work ; and (2) a significant improvement in complex syntax (transfer effects on syntax). This study did not include an active control group of participants receiving an alternative training regimen, since our previous studies with children with DLD or typical development . Finally, participants completed a long-term follow-up three months after training. We predicted that 4) potential effects on WM, syntax and attention would be maintained after three months (long-term effects).The current study also included attentional measures, in light of the observation that WM is closely linked to the attentional system  abilities were established via pre-tests 1 week prior to the first training session, comprising two 45-min sessions to assess memory, syntactic and attentional skills. Participants then completed the 8-week intensive training regimen of three 30-min sessions per week for a total of 12 h. Tests were completed again one week after training (T2) with exactly the same structures but different items, matched in frequency, length and complexity, to avoid practice effects. There were two different test versions, A and B, and administration order was counterbalanced such that half of the participants completed version A pre-tests and version B post-tests, and the other half completed the opposite order. Post-tests (T3) were administered three months after T2, to assess the long-term stability of effects. Participants completed the same version (A or B) at T3 that they had completed at T1, five to six months earlier. As indicated on Fig. Baseline T abilitieM = 8;8, SD = 1;8) from speech-language centers and specialized schools in the Geneva and Paris regions, which we had contacted directly to present the study and its clinical objectives. This age range was chosen since it corresponds to the ages of children for whom relationships between WM and complex syntax had previously been identified  with ASD ages 5 to 11 (Inclusion criteria were: (i) meeting DSM-5  = 4.52, p < 0.001. Appendix A provides these results in detail.Nonverbal skills were age-typical overall; 14 children had above-average Raven’s scores. In expressive syntax, all children obtained low scores, with 20 scoring more than 2 SD below the mean. All children obtained low complex-span WM scores (> 1 SD below the mean for their age); simple span scores were more variable, and the difference between simple and complex spans was significant, This study was approved by the Ethics Committee of the Faculty of Psychology and Educational Sciences of the University of Geneva as well as from the Geneva Cantonal Ethics Commission and was also declared at ‘La Commission Nationale de l’Informatique et des Libertés (CNIL)’ in France. Parents provided written informed consent for participation.Before and after training, children completed tests assessing WM with three simple span tasks and two complex span tasks, see Table Before and after training, children completed three measures assessing syntactic knowledge, shown in Table wh-fronted object (1 and 2) or wh-in situ questions (3), which are less grammatically complex but very frequent in spoken French.Qui pousses-tu?” (‘Who push you’).“Qui tu pousses?” (‘Who you push’).“qui?” (‘You push who’).“Tu pousses In this task, adapted from Jakubowicz , the exprd person accusative clitic (e.g. what is the doctor doing with the boy? Expected response: “il le pèse” ‘he’s weighing him’). Each version included 12 trials. Responses typically contained (i) the target accusative clitic, (ii) a lexical unit (il pèse le garçon ‘he’s weighing the boy’); this response is grammatical but infelicitous as it unnecessarily repeats the full lexical unit; (iii) a clitic with a gender error (ex: *il la pèse ‘he’s weighing her’), or (iv) an omitted clitic (*il pèse ‘he’s weighing’).In this task adapted from Tuller et al.  and Dela(4)‘The teacher sees the boy who is reading a book about Christmas’La maîtresse voit le garçon [qui lit un livre sur Noël](5)‘It’s a cat that all of the children pet after school’C’est un chat [que caressent tous les enfants après l’école](6)‘I think that the girl prefers the dog that she colored’Je crois [que la fille préfère le chien [qu’elle a colorié]](7)‘He thinks that she says that the boy hates the girl who is crying’Il pense [qu’elle dit [que le garçon déteste la fille [qui pleure]]]To test the ability to repeat syntactically complex sentences, we used a sentence repetition task created by Delage and Frauenfelder  that reqAs the task progressed, the structures the children were asked to repeat became increasingly more complex. Scoring for the sentence repetition task considered (i) the number of syllables which were correctly repeated, disregarding possible mispronunciations; (ii) inclusion of the target structure (e.g. an object relative), and (iii) inclusion of the expected degree of embedding . Considerations (ii) and (iii) focused only on syntactic properties. Thus, if the response included the correct structure and/or the expected level of embedding, it was scored as correct even if the response included an incorrect word . Responses were digitally recorded. Trained research assistants transcribed and coded all responses, which were reviewed in full by the first and the third authors.Complementation skills were assessed using a task adapted from De Villiers and Pyers . ChildreVisual selective attention capacity was assessed via the Sky Search task  if it appeared one, two or three trials previously. In the dual-task processing activity, the child had to retain an ordered list of familiar auditory stimuli  while simultaneously performing a secondary task .The WM training was delivered via iPad using the same After verifying that variables met standard assumptions of normality and heterogeneity, we checked for correlations between WM scores and syntax, as well as correlations of WM/syntax to clinical variables . As a first step in the analyses, we created composite variables for WM, syntax, and attention measures . These composites were subjected to repeated-measures ANOVAS, to test for a main effect of the intervention  on WM (direct effects) and syntax and attention (transfer effects). Those composite measures that showed significant change over time were further subjected to repeated-measures ANOVAs to compare performance from T1 to T2 (intervention effects) and from T1 to T3 (long-term effects); we also conducted exploratory analyses of individual WM, syntax, and attention measures, to identify specific domains where the intervention had the greatest impact. Finally, additional analyses probed clinical and cognitive predictors of changes in WM and syntactic ability.Z-score) composite scores for simple and complex spans; each measure contributed equally to the composite. The simple span composite was the average of forward digit recall and serial order word span Z-scores; these individual measures were highly correlated, r(30) = 0.60, p < 0.001. Nonword repetition  was excluded from this composite, as it was uncorrelated with serial order word span, r(30) = 0.29, p = 0.11. The complex span composite was the average of backward digit recall and counting span z-scores, which were also highly correlated, r(30) = 0.47, p = 0.006. Results showed that the complex but not simple span composite correlated with all syntactic measures; see Table Z-scores of (1) Elicited production of root questions, (2) Elicited production of accusative clitics, (3) Sentence repetition, and (4) Comprehension of complement sentences. The attention composite was calculated as the average of the Z-scores of (1) Sky Search and (2) Opposite Worlds tasks.Before exploring the effects of WM training, we ascertained whether WM and syntax were correlated at T1-Baseline, as in previous studies  = 27.89, p < 0.001, Cohen’s f = 0.93, and complex WM composite scores, F = 185.89, p < 0.001, Cohen’s f = 2.48, showed significant main effects of training with large effect sizes. As such, we performed follow-up analyses to compare T1 with T2, for each of the WM measures; scores and statistics are presented in Table To test whether WM training led to significant improvements in WM performance, we performed an initial repeated-measures ANOVA on T1, T2 and T3 scores. Both the simple WM composite scores, F = 98.16, p < 0.001, Cohen’s f = 1.86. As such, we performed follow-up analyses to compare T1 with T2, for each of the syntax assessments; scores and statistics are presented in Table wh-in situ items, showed significant change, t(29) = − 2.2, p = 0.03, d = 0.4. There was no significant change in wh-fronted questions or object clitic productions. For complex sentence repetition, the number of correctly repeated syllables significantly improved from T1 to T2, with a medium-to-large effect. This result cannot only be explained by better memory, because repetition accuracy for simple sentences  showed no significant improvement, t(29) = − 1.2, p = 0.2. The percentage of sentences in which the degree of embedding was respected, independently of the lexical units, also improved. Finally, the children also showed a significant improvement in complement sentence comprehension between T1 and T2.Repeated-measures ANOVAs tested whether training effects transferred to syntax. An initial repeated-measures ANOVA on T1, T2 and T3 composite syntax scores revealed a significant effect of training with large effect size, F1, 2 = 98.16,F = 442.31, p < 0.001, Cohen’s f = 4.04. As such, we employed a series of repeated-measures ANOVAs, detailed in Table To test the hypothesis that WM training would boost selective attention, processing speed and attention shifting, we conducted a third repeated-measures ANOVA on the composite T1, T2 and T3 attention scores; this revealed a significant effect of training, with a large effect size, Of the original sample of 30, 26 children with ASD were retested at T3, three months after the posttest. Repeated measures ANOVAs with time  as a within-subjects factor revealed significant main effects of time were observed for each of the WM tasks; see Table The long-term stability of transfer effects was calculated for those measures of syntax and attention for which a significant T1-T2 increase was observed. As indicated in Table p’s > 0.10.The results revealed a significant impact of WM training on WM itself (direct effects) and on attention and syntax (transfer effects); improvements in the latter were in the moderate range. Given the heterogenous nature of ASD, it was important to test whether improvements were observed for all participants. We calculated gains for the primary measuresMagic Memory  and all measures of syntax in our participants with ASD, replicating previous results . Nevertheless, such direct effects have not been consistently observed  about pictured scenes. While the social demands are highly structured and relatively small, participants must listen and respond to the examiners; this could be a more difficult task for children with ASD , these studies did not assess attentional abilities and were therefore unable to test for the transfer effects on attentional tasks that we obtained in the current study. Future studies should also evaluate these transfer effects in children with DLD. Similar effects would be expected, consistent with studies linking attentional resources to the processing of complex syntax in children with DLD  were retested three months after training to assess long-term effects. Results showed that improvements were maintained in all WM tasks except backward digit recall, for which performance decreased from T2 to T3 . This task requires reversing the order of previously heard stimuli (digits). The training program was similar, but involving color names rather than digits. It is possible that the extensive overlap between tasks boosted the progress observed between T1 and T2, and that these gains were not maintained without practice . Findings of this study provide further evidence in support of the efficacy of WM training in multiple domains, and fall within an ‘evidence-based practice’ framework, which emphasizes the role of research findings in clinical decision-making Below is the link to the electronic supplementary material.


At optimised conditions, the PDFe/Al showed ≥99% removal efficacy for CR dye and an exceptionally high Langmuir adsorption capacity of 411 mg·g−1. Furthermore, a diffusion-limited adsorption mechanism was observed, with two distinct surfaces involved in the adsorption of CR from an aqueous solution. It was determined that the adsorption of CR induced internal strain and deformation within the matrices and interlayers of the PDFe/Al which resulted in a marked increase in the adsorbent pore surface area and pore volume. The remarkably high adsorption capacity could be attributed to the high surface area. A regeneration study showed that the adsorbent could be reused more than four times for the adsorption of CR. The findings from this study demonstrated the feasibility of recovering valuable minerals from toxic and hazardous AMD and demonstrated their potential for the treatment of industrial wastewaters.This study presents the first known exploration of Congo red dye (CR) adsorption by a polycationic Fe/Al Di-metal nanostructured composite (PDFe/Al) synthesised using Fe(III) and Al(III) recovered from authentic acid mine drainage (AMD). The PDFe/Al successfully removed CR from the aqueous solution. The mineralogical, microstructural, and chemical properties of the synthesised PDFe/Al adsorbent (before and after adsorption) were studied using state-of-the-art analytical instruments. The optimum conditions were observed to be 100 mg·L Environmental contamination by coloured, acidic, and metalliferous effluents has been an issue of topical concern globally ,2. This Due to its stellar tenacity, chemical stability, and strong adhesive properties, Congo red dye (CR) has been widely used in printing, clothing, pulp, textile, chemical, and paint industries . The priTo counter toxicological effects, conform to regulatory requirements, and avert potential ecological degradations, a number of technologies have been developed for the removal of CR, including adsorption , precipi2Cr2O7), caustic soda (NaOH), sulphuric acid (98.5% H2SO4), and hydrochloric acid (37% HCl). CR dye stock solution was prepared using CR dye powder. Aqueous samples and solutions were prepared using ultrapure water (18.2 MΩ-cm) prepared using an Elga PURELAB® Flex . Experimental glassware was carefully and thoroughly cleaned before and after every use to avoid contamination.Raw AMD was collected from a coal mine in the Mpumalanga Province of South Africa. All chemicals were purchased from Sigma-Aldrich  and used as obtained without any further purification: potassium dichromate salt . The recovered material was then crushed into a fine powder in a vibrating ball mill  at 700 rpm before being calcined at 800 °C. The samples were then sieved through a 32 μm perforated sieve  to obtain the necessary particle size, and they were stored in a plastic “zip-lock” bag to preserve them from the environment until they were usedThe synthesis of PDFe/Al followed the same method as previously described . To pref−1. The stock solution was made in a 1000 mL volumetric flask that had been filled to the specified level. For each adsorption experiment, fresh aqueous solutions were made from the stock solutions.An amount of 1 g of CR dye was dissolved in 1 L ultrapure water (18.2 M-cm) to make 1000 mg·LA batch-experimental approach was adopted for the removal of CR dye from aqueous solutions using the synthesised PDFe/Al. Different parameters were studied for the adsorption of CR dye, such as initial concentration, adsorbent dosage, agitation time, temperature, and initial solution pH, where 500 mL of the CR dye solution was loaded in individual volumetric flasks. For quality control and quality assurance, all experiments were conducted in triplicate and the data were reported as means. The effects of the different parameters on the adsorption process were studied as summarised in −1) on the adsorption process was studied by preparing fresh aqueous solutions from concentrated 1000 mg·L−1 CR dye stock solutions. The effect of PDFe/Al dosage on CR dye adsorption was studied by varying different dosages of the adsorbent (0.1–5 g) and agitating them with the respective adsorbate solutions. The effect of agitation time was studied by conducting the batch adsorption experiments for CR dye adsorption for different time intervals (1–60 min). The effect of initial solution pH was studied by adjusting the pH of CR dye solutions with 0.1 M NaOH and/or 0.1 HNO3 to a desirable level (pH of 2–10). The effect of temperature was studied by conducting the batch adsorption experiments in a thermal shaker/incubator under different temperatures (25–65 °C). Concentrations of CR dye were analysed before and after the experiments.As depicted in To control for potential spectrophotometric interference resulting from the adsorbent matrix, the adsorbent was agitated in 500 mL distilled water overnight and, after the removal of the adsorbent, the eluent was analysed spectrophotometrically. No quantifiable change to the medium was measurable and, therefore, it was concluded that the adsorbent matrix would not cause spectrophotometric interference during the experiment.®, Columbus, OH, USA) FiveGo EC/TDS/Salt/Temperature portable multimeter was used to measure electrical conductivity (EC), total dissolved solids (TDS), and salinity. The absorbance of the CR dye was measured using a WPA Lightwave II UV/Visible spectrophotometer .The pH levels of the aqueous solutions were measured using a Thermo ScientificTM Orion 3 Star portable pH meter before and after the adsorption experiments . A Mettler Toledo . The synthesised PDFe/Al and subsequent residues’ Brunauer–Emmet–Teller (BET) surface area and BJH pore size were determined using BET equipment and a Micromeritics VacPrep 061 degassing system . The thermal stabilities of the synthesised PDFe/Al and residual products were determined using a Thermo Gravimetric Analyser (TGA)  in ambient air at a flow rate of 50 mL·min−1 and a heating rate of 10 C·min−1.X-ray diffraction (XRD) was used to determine the mineralogical composition and pattern of the Fe/Al and residual products, which was recorded using a Panalytical X’Pert PRO  equipped with a Cu-K radiation source. The Origin 2021b software program was used to deconvolute the XRD signal peak . High-resolution scanning electron micrographs (HR-SEM-EDX) were used to examine the surface morphology and composition of solid materials . A Perkin-Elmer Spectrum 100 Fourier Transform Infrared Spectrometer (FTIR) with a Perkin-Elmer Precisely Universal Attenuated Total Reflectance (ATR) sampler was used to examine the metal functional groups in solid samples (Perkin-ElmerThe Smiiklas method  was usedeQ (mg·g−1) is the adsorption capacity, C0 (mg·L−1) is the starting concentration of CR dye, eC (mg·L−1) is the equilibrium concentration of CR dye, V (L) is the volume of the CR dye solution, and m (g) is the dosage of PDFe/Al, respectively.Equation (1) was used to evaluate the adsorption capabilities of PDFe/Al for the removal of CR dye from aqueous solutions:eR denotes the PDFe/Al removal effectiveness, C0 denotes the initial CR dye concentration (mg·L−1), and eC denotes the equilibrium CR dye concentration (mg·L−1).Equation (2) was used to calculate the elimination effectiveness of CR dye by PDFe/Al:−1 CR dye solution in a 250 mL Erlenmeyer flask for 90 min in a batch experiment. After equilibration, the residue was centrifuged to separate it from the supernatant. To eliminate excess CR dye ions, the recovered residual material was washed five times with 250 mL of ultrapure water and dried. After that, 250 mL of 0.1 M HNO3 was added to the dried sample at room temperature. The HNO3 extract that resulted was collected and analysed for CR dye ions. Equation (3) was used to calculate the regeneration percentage:desC (mg·L−1) is the concentration of CR dye ions in the desorption eluent and C0 (mg·L−1) is the initial concentration of CR dye ions.An approach described by Kumari et al.  was usedA Fourier Transform Infrared Spectrometer (FTIR) was used to characterise the functional groups of the synthesised PDFe/Al before and after CR dye adsorption, as shown in −1, while at 1668 cm−1, HOH stretching is observed [−1, likely an indication of S=O- stretching, is observed. S=O- was also observed in the spectrum for the pure CR at 1278, 1177, and 1041 cm–1 [−1, which could be an indication of –OH stretching in α-FeOOH [−1 The wavebands at 3740 and 2966 cm−1 could also indicate the stretching of the –OH group. The AlOOH band is observed to have stretched at 630 cm−1 [As depicted in observed . Further041 cm–1 . A small α-FeOOH . Further630 cm−1 .X-ray diffraction (XRD) was used to characterise the mineralogical composition of the synthesised PDFe/Al before and after CR dye adsorption, as shown in As illustrated in From the relative positions of the fitted Gaussian peaks—shown in Scanning electron microscopy (SEM) was used to characterise the morphology of the synthesised PDFe/Al before and after CR adsorption, as shown in As illustrated in An energy dispersive X-ray (EDX) was used to characterise the mapping of the elemental composition of the synthesised PDFe/Al before and after CR adsorption, as shown in As illustrated in Thermo-gravimetric analysis (TGA) was used to determine the thermal stability of the synthesised PDFe/Al before and after CR adsorption, as shown in As illustrated in BET surface area analysis was used to determine the surface area and porosity properties of PDFe/Al before and after CR dye adsorption, as illustrated in 2.g−1 to 134 m2.g−1. Further analysis of the pore volume ; however, there is much uncertainty in the future observations relative to CR dye adsorption application, as seen in the very large 95% prediction interval due to the model’s dependence on initial adsorbate concentrations, thus limiting the application of the PFO kinetic model. In 2 = 0.997) for the adsorption of CR dye. The TPA model provides insight into the mechanism of adsorption as well as the relative dominance of the adsorption kinetics on the adsorbent surface. Similar to what was proposed by Wang et al. [e)(D [eD = 3.49 × 10−11 m2·s−1) and the molecular diffusivity of CR (2.16 × 10−9 m2·s−1 [2 = 0.999), and the findings were good, where eD1 = 4.30 × 10−12 m2·s−1 and eD2 = 6.47 × 10−15 m2·s−1, supporting the observations from the Crank model  . The res9 m2·s−1 ) showed 9 m2·s−1 . The datnk model d of signAdsorption isotherms for the adsorption of CR dye by PDFe/Al were studied to elucidate the different mechanisms governing the adsorption process. The isotherm models are summarised in 2R < 0.99 and a root-mean-square error RMSE >12.7 mg·g−1, indicating that no single isotherm model was able to conclusively predict the equilibrium data. However, sufficiently good fits were obtained to provide some insights into the mechanisms responsible for adsorption.max,LQ) of 411 mg·g−1 was predicted. This is exceptionally high when compared to other reported studies. Of further interest is the value LR = 0.775, indicating favourable adsorption conditions—0 < LR < 1 is considered favourable for adsorption, L >R 1 unfavourable for adsorption, LR = 1 linear adsorption, and L =R 0 irreversible adsorption [2 = 0.922.sorption . This maFn) gives an indication of the favourability of the adsorption—nF < 1 is considered unfavourable, Fn = 1 designates linear adsorption, and Fn > 1 shows favourable adsorption [Fn =1.99 indicates highly favourable adsorption.sorption ,64. In tmax,Q1 = 33.0 mg·g−1 and max,Q2 = 1270 mg·g−1 for two surfaces, giving a total maximum adsorption of 1303 mg·g−1—significantly greater than that previously reported for CR adsorption. However, since the total maximum adsorption capacity is a gross extrapolation of the experimental data, the more conservative Langmuir adsorption capacity  was adopted as the official maximum adsorption capacity for the study. What is of most interest is that the two-surface Langmuir model proposes that the adsorbent surface consists of two separate adsorption surfaces with different energies and sorption properties on which parallel adsorption takes place [max,Q1 = 33.0 mg·g−1, while the second adsorption surface exhibited reversible adsorption with a remarkably large max,Q2 = 1270 mg·g−1. This is supported by the observations that the L,R1 = 0.0145 indicates nearly irreversible adsorption, while L,R2 = 0.95 indicates highly favourable adsorption conditions [The results from this model were markedly better than that for the single surface Langmuir model with es place  . The adsorption energies for RD and AD isotherms were determined and found to be DRE = 15.0 kJ·mol−1 and DAE = 14.7 kJ·mol−1. The values for the adsorption energies indicate a diffusion-controlled adsorption mechanism—energies >16 kJ·mol−1 designate chemisorption, energies <8 kJ·mol−1 correspond to physisorption, and energies between 8 and 16 kJ·mol−1 indicate mass transfer limited diffusion [iffusion max,SQ = 1350 mg·g−1, SK = 204.15 and Sn = 1.93 for the Sips isotherm. According to Keren et al. [Sn. The Sips isotherm reduces to the Langmuir isotherm for nS = 1, while a value for nS > 1 results in a sorbate–sorption site interaction characterised by a Gaussian distribution (known as the Langmuir–Freundlich model). A value for nS < 1 indicates a cooperative reaction between sorption sites and n sorbate molecules; this type of adsorption is characterised by sigmoidal adsorption isotherm data. The Sips isotherm model further reduces to the Freundlich model for Sn > 1 and at low adsorbate concentrations (when Sn = 1.93 shows that favourable adsorption conditions are present (similar to the Freundlich isotherm). The Sips isotherm model links strongly to the two-surface Langmuir model which predicts a heterogeneous surface with two main surface sites showing differing adsorption characteristics.n et al. , the SipmaxQ for both the two-surface Langmuir and Sips models are >1300 mg·g−1, which is remarkably high for CR adsorption. However, as indicated before due to the uncertainty inherent in such significant extrapolation, the max,LQ was adopted as the official maximum adsorption capacity of the study.The values for the max,LQ) and the favourability of adsorption (Fn > 1). This is especially pertinent considering that the only materials to outcompete the current materials in terms of max,LQ were synthesised using laboratory-grade chemicals, whereas the PDFe/Al was synthesised using bona fide AMD. In addition, the materials showing higher adsorption favourability (higher Fn) displayed significantly lower maximum adsorption capacities, indicating that PDFe/Al represented a highly desirable combination of high adsorption capacity and favourability towards CR adsorption.As illustrated in The material showed the potential to be reused more than four times for CR dye adsorption. It should be noted that even during the sixth regeneration cycle the % desorption ≈ 95%, indicating a significantly stable and regenerable adsorbent for industrial application.The proposed adsorption mechanism for CR adsorption is summarised in −11 m2·s−1 being two orders of magnitude smaller than the molecular diffusivity of CR of 2.16 × 10−9 m2·s−1 [DRE = 15.0 kJ·mol−1 and DAE = 14.7 kJ·mol−1, both indicating a diffusion-controlled adsorption mechanism [−1 and one reversible with and adsorption capacity of 1270 mg·g−1. This multi-surface characteristic is supported by two-phase adsorption kinetics, showing a fast and slow adsorption process taking place in parallel. Finally, it was observed that the adsorption of CR on the adsorbent surface induced significant internal strain within the adsorbent, which resulted in deformation, cracking, and subsequent increased adsorption area and volume within the adsorbent. This increased area, coupled with electrostatic interactions and additional covalent/donor–acceptor interactions between the surface of the adsorbent and CR dye ions, could explain the high adsorption of CR dye ions by PDFe/Al [The adsorption of CR dye by PDFe/Al appears to be predominantly diffusion-controlled, with the greatest predicted effective diffusivity of 3.49 × 109 m2·s−1 . This isechanism . The dif PDFe/Al .This study has demonstrated multiple technical feasibilities in the use of the synthesised nanocomposite for the removal of CR dye. Henceforth, upscaling this technology to a viable industrial scale could be possible, specifically considering the amount of mine water being generated in South Africa. Furthermore, findings from our laboratory assays have confirmed the efficacy of the synthesised PDFe/Al; the next stage should be to pilot the application for the removal of CR using real industrial effluents.Column tests will be carried out to assess the developed materials’ ability to remove CR continuously.A techno-economic analysis of the suggested material should be performed to determine the technology’s economic viability.The doping of the material should be considered to boost its adsorption capacity, hence increasing the proposed materials’ adsorption efficiency.A life-cycle analysis of the proposed system will be used to determine the environmental sustainability of the technology.Future work and Recommendations:bona fide AMD. The study successfully demonstrated the highly effective removal of CR from aqueous solution by PDFe/Al. The optimised conditions for CR dye adsorption were found to be: 100 mg·L−1 initial CR dye concentration, 1 g of PDFe/Al in 500 mL adsorbate solution, pH 3–8, 20 min agitation, and a temperature of 35 °C. The synthesised PDFe/Al had a Langmuir adsorption capacity of 411 mg·g−1 CR dye with >99% removal of CR—one of the highest for CR adsorption. The adsorption mechanism was found to be significantly diffusion-limited, with a dual-site adsorption surface responsible for adsorption. It was further found that CR adsorption induced strain and deformation within the PDFe/Al, resulting in internal crack formation and a consequent increase in pore surface area (37.58 m2.g−1 before adsorption vs 134.46 m2.g−1 after adsorption) and pore volume (0.0621 cm3. g−1 before adsorption vs 0.1137 cm3. g−1 after adsorption). This remarkable increase in the surface area could provide an explanation of the notable adsorption capacity measured. Finally, it was determined that the PDFe/Al could be regenerated and reused more than four times with minimal loss in adsorption efficiency. The current study successfully valorised and beneficiated AMD and provides an avenue to foster and grow the circular economy in wastewater treatment.This double-edged study presents the first known investigation into the adsorption of CR by PDFe/Al synthesised from


In recent years, diabetes has become a global health problem that creates a tremendous economic burden for many countries. Clinical trials evaluating the hypoglycemic effects of sesame consumption have produced conflicting results. This systematic review and meta-analysis was conducted to evaluate the effectiveness of sesame as a popular natural herb on glycemic indices in adults.  The search for related articles in PubMed, Scopus, Google Scholar, and Cochrane library was conducted through May 2021. Results were reported as weighted mean differences (WMD) with 95% confidence intervals (CI) using a random-effects model. I2 = 97.6%; 95% CI ), and hemoglobin A1c (HbA1c): , I2 = 0%; 95% CI ) as compared to the control group.  A total of 605 studies were identified through online searching, and a total of eight RCTs representing 382 participants were included in this study. The meta-analyses revealed that sesame consumption significantly decreases serum fasting blood sugar (FBS): ,  This study provides evidence of the hypoglycemic effects of sesame consumption, particularly in diabetic patients. Additional RCTs on sesame and its preparations should be conducted in different populations to increase generalizability. Diabetes mellitus is one of the most common metabolic disorders, affecting over 420 million individuals worldwide . This coSesamum indicum L.) has been used as a traditional plant-based therapy worldwide, most commonly in Asian regions [α-tocopherol, polyunsaturated fatty acids (PUFAs), monounsaturated fatty acids (MUFAs), and fiber [Sesame  guidelines for this study  Supplem.PubMed/Medline, Cochrane Library, Google Scholar, and Scopus were searched to locate relevant RCTs published through May 2021. The search strategy was prepared using medical subject headings (MeSH) and dependent keywords as follows: ((“Sesame” OR “Sesame Oil” OR “Sesame Seed” OR “Sesamum”) AND (“Blood Glucose” OR “Insulin” OR “Glycated Hemoglobin A” OR “Insulin Resistance” OR “Diabetes Mellitus” OR “Glucose” OR “Blood Glucose” OR “Insulin Resistance” OR “diabetes” OR T2DM OR “hyperglycemia” OR “hyperglycemia” OR “HbA1c” OR “insulin sensitivity” OR “HOMA” OR “glucose homeostasis “OR “insulin secretion” OR “beta-cell function” OR “glycemic control” OR “glucose tolerance” OR “glucose metabolism”)). No restrictions were used when searching the databases. To ensure all related articles were identified, we checked the reference list of all qualified studies and related reviews.Two investigators independently browsed the title and abstract of every article and included them if they met the inclusion criteria. To be included, the studies had to be RCTs  examining the effect of sesame seed and its preparations (oil/flour/supplement) on glycemic indices such as serum fasting blood sugar (FBS), hemoglobin A1C (HbA1C), insulin concentrations, insulin resistance (HOMA-IR: homeostasis model assessment of insulin resistance), and insulin sensitivity (QUICKI: quantitative insulin sensitivity check index).Studies that used other compounds in addition to sesame only in the intervention group, studies without a control group, and studies with the same populations were excluded.Two researchers independently extracted the following data from every eligible article: author's name, publication year, country, sesame source and quantity, duration of intervention, participant's information , study type, and glycemic indices values (mean and standard deviation) before and after the intervention. Discrepancies were solved by consulting with the principal investigator.Risk of bias was evaluated by two researchers independently using the Jadad checklist . This scQuality of the evidence was evaluated using the Grades of Recommendation, Assessment, Development, and Evaluation (GRADE) tool . This toR value when calculating SD change [I2 index and its confidence interval [I2 values of 25–50%, 50–75%, and >75%, respectively [P < 0.05.Effect sizes were reported as weighted mean differences (WMD) and 95% confidence intervals (CI) and were obtained by combining effect sizes of mean (SD) changes of glycemic control biomarkers using random-effects models. Due to the small number of the studies in these analyses, the Hartung–Knapp adjustment was also used . We calcD change . Betweeninterval . Low, moectively . PredeteA total of 605 articles were entered into Endnote for screening, as shown in General characteristics of the included studies are reported in I2 = 97.6%; 95% CI ) ) . The res sources .I2 = 0%; 95% CI   , and HarI2 = 98.3%%; 95% CI , Hartung–Knapp adjustment analysis 7.52 IU/ml; 95% CI , I2 = 98.3%).Meta-analysis of three studies , 15, 28 The sensitivity analysis found that none of the studies dramatically influenced the overall findings of FBS, HbA1C, and insulin level , HbA1c (P=0.22), and insulin (P=0.98)  in the A1C percentage of 0.5% is considered clinically significant . This isWe also found that serum FBS and HbA1C declined among people who consumed sesame. This is consistent with findings from an in vitro study on diabetic rats, which produced evidence that sesamin leads to a significant improvement in the serum FBS and HbA1c in a dose-dependent manner by promoting glycogen synthesis in the liver . SimilarThese findings differ from a randomized, triple-blind, three-way crossover trial by Raeisi-Dehkordi et al. with 95 subjects who have type 2 diabetes (18–60 years). These patients substituted sesame oil, canola oil, or sesame-canola oil for their regular dietary oil for 9 weeks with a 4-week washout period. When sesame oil or sesame-canola oil was compared to canola oil, no significant effect on FBS, insulin, and HOMA-IR was observed . Howeverα-tocopherol (40 mg/100 g), PUFA (43%), and MUFA (40%) [These discrepancies might also be explained by the difference in bioavailability among lignan compounds in sesame products . Most of the studies in this analysis used sesame oil. Sesame oil contains greater quantities of lignan (11.51 mg/g)  than sesFA (40%) . EpidemiFA (40%) . High amFA (40%) .It is also important to note the lack of consistency regarding placebo substances used for comparison to sesame. These oils  contain different levels of PUFA, MUFA, and other biological compounds which can positively influence glycemic markers. It is challenging to quantify the actual effect of sesame on glycemic indices above that of a placebo due to the known therapeutic effect of these oils.β-cell destruction and elevating insulin sensitivity [The hypoglycemic effects of sesame may be explained by multiple potential mechanisms. Sesamin can inhibit the augment of blood glucose by playing a role in increasing glycogen production . It has sitivity , 39. Thesitivity . Sesame sitivity .This study has several limitations. The small number of trials identified in the literature suggests that the present systematic review and meta-analysis results may be biased by the sample size. Similarly, eligible studies were found to be heterogeneous regarding sesame preparation, placebo content, duration of intervention, and sample size.The overall result may have been influenced by the differences in dosing and sourcing of sesame. Subgroup analysis to identify dose-dependence or contrast groups by dosing was not feasible due to variations in the sesame preparations and the difficulty of obtaining accurate dose conversions.Additionally, several of the studies in this analysis was found to be of poor quality, limiting the generalizability of these findings. Several trials failed to sufficiently describe the process of randomization, allocation concealment, and blinding. There is also a geographic limitation with most of the eligible studies having been conducted in Asian countries. Finally, the majority of trials did not control for the dietary changes of participants during the intervention, which may have influenced the outcome.The protocol of this study has not been registered in PROSPERO. However, PROSPERO and all relevant databases were comprehensively searched to confirm that no systematic review and meta-analysis has yet been conducted.This systematic review produces evidence that sesame consumption could decrease glycemic indices, especially in diabetic patients. This study finds that sesame is a safe and effective complementary approach to improve hyperglycemia in clinical practice. However, the small numbers of related RCTs and high amounts of heterogeneity among these studies limit generalizability. We recommend that additional high-quality RCTs be conducted on participants with different health conditions, varying duration of intervention, and multiple kinds and dosages of sesame consumption to confirm our findings.


It was cultivated from the bud-mutation of “Yinghong 9” and has a very low level of chlorophyll during young shoot development. Due to chlorophyll being involved in carbon fixation and assimilation, the changes in photosynthesis might potentially affect the accumulation of flavor metabolites, as well as the quality of “Huangyu” tea. Although “Huangyu” has a golden yellow color and high amino acid content, the mechanism underlying the formation of leaf color and drinking value remains unclear. The widely targeted metabolomics and GC-MS analysis were performed to reveal the differences of key metabolites in fresh and fermented leaves between “Yinghong 9” and “Huangyu.” The results showed that tea polyphenols, total chlorophyll, and carotenoids were more abundant in “Yinghong 9.” Targeted metabolomics analysis indicated that kaempferol-3-glycoside was more abundant in “Yinghong 9,” while “Huangyu” had a higher ratio of kaempferol-3-glucoside to kaempferol-3-galactoside. Compared with “Yinghong 9” fresh leaves, the contents of zeaxanthin and zeaxanthin palmitate were significantly higher in “Huangyu.” The contents of α-farnesene, β-cyclocitral, nerolidol, and  Camellia sinensis) is an important economic crop in south China. The sensory quality of tea products depends on their chemical compounds, although it is still challenging to illustrate. Color and flavor are two important sensory quality indicators for tea products. The leaf colors of tea cultivars often have severely effects on appearance and flavor which involved in manufacturing suitability of tea leaf  is a large leaf tea cultivar selected from Yunnandaye tea, it is a suitable cultivar for the manufacture of black tea because of its higher tea polyphenol content and flowery-aroma flavor. “Huangyu” is a yellow leaf mutant that bred from bud mutation of “Yinghong 9,” and it has a relatively low level of chlorophyll than the parent plant  in August 2020. The mutant materials (“Huangyu”) were collected from the mutant plant, which was grafted on the parent plant (“Yinghong 9”). The tea leaves were equally divided into three portions. The samples were prepared by black tea processing described as follows: the fresh tea leaves were subjected to indoor natural withering at 25–27°C by spreading on bamboo sieves for 15 h. After withering, the leaves were rolled by a mini rolling machine for 60 min. Then the leaves were put into a bamboo-basket to ferment for 6 h at 25–27°C with a relative humidity of about 85%. During the black tea processing, fresh and fermented leaves were collected and immediately fixed with liquid nitrogen. And then the samples were ground to powder and stored in a freezer at −80°C before the following analysis. The water content of these leaves was measured at different stages during the black tea processing.One bud and two leaves of The tea polyphenols content was determined by spectrophotometer method with minor modifications . The absThe determination of total chlorophyll and carotenoids contents were extracted and determined as previously described with some modifications ; The growhere A663, A646, and A470 represent the absorbance at 663, 646, and 470 nm, respectively; where V represents the total volume of extracting solution (ml), and W represents the fresh weight (FW) of the samples (g); The contents of chlorophyll and carotenoids were estimated in mg/g (FW).g for 15 min, the supernatant was filtered with SCAA-104  and analyzed by UPLC-MS/MS.All above frozen samples were freeze-dried by LGJ-10 experimental vacuum freeze dryer . The freeze-dried samples were ground using a MM 400 grinder  with a zirconia bead for 90 s at 30 Hz. The lyophilized sample (100 mg) was extracted with 1.2 mL 70% methanol solution for 24 h at 4°C. During this period, the samples were vortexed  30 s once every 30 minutes for a total of six times to facilitate the extraction. Following centrifugation at 12,000 1) and proprietary database  following the previously operating procedures  ≥ 1 and fold change ≥ 2 or ≤ 0.5. VIP values were extracted from orthogonal partial least squares discriminant analysis (OPLS-DA) results, which were conducted using R package  according to the procedure described by ocedures . Metabol package .The freeze-dried samples were ground  using a MM 400 grinder . Fifty milligrams of ground samples were mixed with an appropriate volume of the extraction solution  with 0.01% butylated hydroxytoluene (g/mL) and internal standard. The mixture was adequately vortexed  at room temperature for 20 min. The mixture solution was then centrifuged , and the supernatant was collected. The samples were repeatedly extracted two times using the same method, and the supernatants were combined and concentrated. The concentration of extracts redissolved in mixture solution . The solutions were filtrated by SCAA-104 filter  before UPLC-MS/MS analysis. Carotenoid identification and quantification analysis were performed by UPLC-MS/MS system at Metware Biotechnology Company . UHPLC Chromatographic and APCI-Q Trap-MS/MS conditions were conducted following the previously stated procedure . CarotenVolatile constituents were extracted according to the previous study  with ming for 20 min at 4°C, and then the supernatant was immediately used for enzyme assays. The enzyme activities of samples were determined according to the protocol of the assay kits, respectively .The carotenoid cleavage crude enzyme extracts were carried out as a method described before . The polpost hoc test. P value less than 0.05 was considered statistically significant. Excel 2010  was applied to drawbar graphs of the experimental data. Flavonol metabolites heat map was prepared using pheatmap in R package (??). The carotenoids heat map was prepared on major bio cloud platform .Statistical analysis was carried out with SPSS software . Significant differences between different groups were determined using Tukey’s p < 0.05) than those in fresh leaves  ≥ 2 or ≤ 0.5) . InteresCompared with “Yinghong 9” fresh leaves, there were 408 differential metabolites in fermented leaves (225 up-regulated and 183 down-regulated), while 439 differential metabolites in “Huangyu” fresh leaves were different from fermented leaves (255 up-regulated and 184 down-regulated) , 3. The O-xyloside was the most abundant detected compound in “Yinghong 9” and “Huangyu” fermented leaves -phytoene, apocarotenal, zeaxanthin, lycopene, violaxanthin and neoxanthin . The typMost of the carotenoids significantly decreased in “Yinghong 9” fermented and “Hongyun” fermented leaves, e.g., β-carotene, lutein, zeaxanthin. On the contrary, some carotenoids significantly increased in fermented leaves, such as antheraxanthin, violaxanthin, and neoxanthin in “Yinghong 9” fermented leaves; lutein caprate and violaxanthin in “Huangyu” fermented leaves. Interestingly, lutein caprate even was not detected in “Yinghong 9” fresh leaves, which markedly increased in “Huangyu” fermented leaves. The reason may be that the lutein caprate is the product of β-carotene degradation in fermented leaves.In order to study the differences of volatile compounds, volatile compounds analyses were carried out in “Yinghong 9” and “Huangyu” fresh leaves and fermented leaves . Twenty-cis-allo-ocimene and 3-Hexen-1-ol, acetate significantly decreased during the fermentation of “Yinghong 9” as well as “Hongyun” cultivars. Some new volatile compounds, e.g., benzaldehyde, benzeneacetaldehyde, linalool oxideI, linalool oxide II,3-hexenyl-α-methylbutyrate, 2-hexenyl isovalerate, geraniol, cis-3-hexenyl hexanoate, cis-hexenyl hexanoate, cis-2-hexenyl hexanoate, α-bergamotene, 6,10-dimethyl-5,9-undecadien-2-one (trans-geranylacetone) and nerolidol were detected in “Yinghong 9” and “Huangyu” fermented leaves. At the same time, the contents of α-farnesene, β-ionone, β-cyclocitral and nerolidol in fermented leaves were significantly higher than those of fresh leaves.As confirmed by previous research, new volatile compounds were generated as the fermentation process proceeds. In the present study, the numbers of volatile compounds increased from 22 (fresh leaves) to 35 (fermented leaves) in “Yinghong 9” . SimilarTo reveal the manufacture suitability of “Yinghong 9” and “Huangyu,” the enzyme activity of polyphenol oxidase β-glucosidase and carotenoid cleavage dioxygenase were measured in their fresh leaves and fermented leaves. The results showed that the activities of polyphenol oxidase, β-glucosidase in “Yinghong 9” were higher than that of “Huangyu,” Similarly, the activity of carotenoid cleavage dioxygenase (CCD) showed also had the same change tendency in “Yinghong 9” and “Huangyu” fresh leaves . The difB-Glucosidase is one of the important enzymes involved in tea aroma compounds formation  leaves, is an important, widely popular non-alcohol beverage due to its health benefit functions, as well as its pleasant flavor. Unlike most other agricultural products, the color of infused leaves is involved in the sensory quality formation of tea. Thus, recently the researchers paid more attention to the study on the relationship of leaf color and characteristic compounds, and its influence on health benefits. In fact, compared to typical green leaves, several albino tea varieties have been successfully developed into new products because of their precious and tasteful quality, e.g., “Anji baicha,” “Yujingxiang,” and “Huangjinya.” Unluckily, tea breeding goals have been limited to a brighter color or better taste. Other valuable traits such as yield, flavor value, and manufacture suitability have long been ignored.Tea, produced from fresh tea -phytoene, β-cryptoxanthin, β-carotene, and neoxanthin were significantly higher in “Yinghong 9.” In comparison, the content of zeaxanthin, and zeaxanthin palmitate were significantly higher in “Huangyu” leaves. Previous studies showed that the changes in the abundances of carotenoids were closely relevant to the expression of carotenoid biosynthesis genes .In plants, zeaxanthin is an essential metabolite of β-carotene. β-carotene generates zeaxanthin through the catalysis of β-carotene hydroxylase (BCH) . SilenciFlavonol glycosides were one of the most important groups of polyphenols besides catechins in tea. Previous studies showed that flavonol glycosides were the critical ingredient of green tea soup, in which they color the tea liquor as yellow or yellow-green pigments . Like caGlucosylation and galactosylation were two competing glycosylation metabolic ways. Simultaneously the content of them varies among different tea varieties. The content of galactosylated flavonol in tea shoots leads to better manufacturing-suitability of green tea. In contrast, the content of glucosylated flavonol was significantly higher in black tea manufacturing-suitability varieties . This imIn this study, the content of kaempferol-3- glucoside in “Yinghong 9” was 1.5 times more than that of “Huangyu.” And the ratio kaempferol-3-glucoside to kaempferol-3-galactoside was lower, which indicates that the glycosylation metabolism of kaempferol in tea plants flowed toward galactosylation, which was beneficial for reducing the content of kaempferol-3-glucoside. So, “Huangyu” is suitable for manufacturing non-fermented tea, e.g., green tea and white tea. Correspondingly, “Yinghong 9” was suitable for manufacturing black tea since it had a high concentration of kaempferol-3-glucoside as well as a lower ratio of kaempferol-3-glucoside to kaempferol-3-galactoside . Liu et Tea aroma quality, which is composed of the volatile compounds that originated during tea processing, is an important quality factor and determines dry tea’s economic benefit. A large number of literatures was concerned about analysis technology, non-carotenoid volatile compounds formation during tea processing, as well as the difference of varieties. The relationship between the carotenoids and their volatile components in yellow leaf tea varieties has received little attention. The formation of volatile compounds from carotenoids comes from the various degradation pathways. Evidence showed that β-ionone, a critical flowery component was the product of β-carotene oxidation degradation, in addition, linalool, nerolidol, β-cyclocitral and geranylacetone were degradation products of other carotenes present in tea . In factAs stated above, β-Glucosidase was involved in tea aroma compounds formation, which could hydrolyze hydrolysis of β-1,4- glycosidic linkages and release free volatile compounds such as linalool, benzaldehyde and methyl salicylate during tea processing . Our stuSimilarly, the content of zeaxanthin, and zeaxanthin palmitate were significantly higher in “Huangyu” leaves, while the contents of nerolidol, α-farnesene and geranylacetone were significantly low in “Yinghong 9” fresh leaves, but the contents of these compounds were higher in ‘Huangyu’ fresh leaves. Correspondingly, the carotenoid cleavage dioxygenase activity was significantly higher in “Huangyu” than in “Yinghong 9” fresh leaves . Due to In this study, analysis of non-volatiles and volatiles was conducted to identify the major metabolite compounds in “Yinghong 9” and its yellow-leaf mutant “Huangyu.” The contents of total carotenoids and chlorophyll revealed that chlorophyll concealed the coloration of carotenoids and led to the formation of normal green leaf in “Yinghong 9.” But for “Huangyu,” being the absence of chlorophyll in leaf, leaf coloration was predominated by carotenoids. The content of kaempferol-3- glucoside in “Yinghong 9” was 1.5 fold more than that of “Huangyu,” and the ratio of kaempferol-3-glucoside to kaempferol-3-galactoside was lower, indicating that “Huangyu” was suitable for manufacturing non-fermented tea, e.g., green tea and white tea, while “Yinghong 9” was suitable for manufacturing black tea. Analysis of carotenoid metabolites revealed that the contents of lutein and α-carotene were significantly higher in “Yinghong 9,” while zeaxanthin was the dominant compound in “Huangyu” leaves. In addition, the contents of nerolidol, α-farnesene and geranylacetone were significantly higher. And the higher activity of β-glucosidase and carotenoid cleavage dioxygenase in “Huangyu” fresh leaves also implied that “Huangyu” was suitable for manufacturing green tea. At the same time, nerolidol and geranyl acetone, the carotenoid degradation products, increased in “Yinghong 9” fermented leaves, which meant “Yinghong 9” was suitable for manufacturing black tea. Our current results provided a basis for understanding the mechanisms of yellow-leaf formation in “Huangyu” and facilitated the effective utilization for particular natural mutants.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.XM designed the experiment. CL and XM carried out most of the experiments. LZ drafted the manuscript. BC and SW did the data analyses. LZ and HW revised and finalized the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


A critical role linking sleep with memory decay and β-amyloid (Aβ) accumulation, two markers of Alzheimer’s disease (AD) pathology, may be played by hippocampal integrity. We tested the hypotheses that worse self-reported sleep relates to decline in memory and intra-hippocampal microstructure, including in the presence of Aβ.APOE ε4 status, and polygenic scores for sleep and AD, respectively.Two-hundred and forty-three cognitively healthy participants, aged 19–81 years, completed the Pittsburgh Sleep Quality Index once, and two diffusion tensor imaging sessions, on average 3 years apart, allowing measures of decline in intra-hippocampal microstructure as indexed by increased mean diffusivity. We measured memory decay at each imaging session using verbal delayed recall. One session of positron emission tomography, in 108 participants above 44 years of age, yielded 23 Aβ positive. Genotyping enabled control for Worse global sleep quality and sleep efficiency related to more rapid reduction of hippocampal microstructure over time. Focusing on efficiency (the percentage of time in bed at night spent asleep), the relation was stronger in presence of Aβ accumulation, and hippocampal integrity decline mediated the relation with memory decay. The results were not explained by genetic risk for sleep efficiency or AD.Worse sleep efficiency related to decline in hippocampal microstructure, especially in the presence of Aβ accumulation, and Aβ might link poor sleep and memory decay. As genetic risk did not account for the associations, poor sleep efficiency might constitute a risk marker for AD, although the driving causal mechanisms remain unknown. Sleep links with memory decay and β-amyloid (Aβ) accumulation, two markers of Alzheimer’s disease (AD) pathology, and a critical role connecting these factors may be played by hippocampal integrity. We performed a longitudinal study testing how self-reported sleep patterns related to changes in memory and intra-hippocampal microstructure, including in the presence of Aβ. We found that worse global sleep quality and sleep efficiency related to more rapid reduction in hippocampal microstructure. In older adults, the relation was stronger in presence of Aβ accumulation. Sleep efficiency related to memory decay indirectly via hippocampal decline. Genetic risk for sleep efficiency and AD did not explain the results, which suggest that poor self-reported sleep efficiency might constitute a separate risk marker for AD.Individuals with sleep disturbances have increased risk for Alzheimer’s disease (AD) , and accAPOE ε4 genotype . I. Ip.adjua v4.9–8 ). To illSummary measures of the PSQI variables can be found in p < .001, R2 =.041, p = .661), and when adding baseline IQ to the covariates (memory change p < .001).p (pFDR) < .05, R2 =.024, p (puncorr) = .012), and (2) sleep efficiency . To further assess this lack of sleep × age interactions, we calculated the 95% confidence intervals (CIs) for the regression coefficient of this interaction term across the seven models. The minimum and maximum of the CIs were −0.034 and 0.036, respectively, indicating a maximum decrease or increase in hippocampal MD of ~0.035 SPC for each unit increase in sleep × age variables. We then calculated the expected annual change in hippocampal MD, using a linear model with interval in years between baseline and follow-up MRI as independent variable, correcting for baseline age. This model yielded a regression coefficient of expected annual change of 0.12 (SPC). As 0.036 is one third of 0.12, we are confident that the effect of a unit increase in sleep on aging is less than half the effect of one year of hippocampal MD change. The null findings are also in line with a recent study ), Z = −2.70, p = .007). The partial correlation obtained in the main sample was within this confidence interval, suggesting a relation between sleep efficiency and memory change may exist, but needing a larger sample to detect it.Poor sleep efficiency was not strongly related to memory decline . The median direct effect estimate, from sleep efficiency to memory change controlling hippocampal MD change, was −1 (p = .222). These results suggested that hippocampal MD change partly mediated the relation between sleep efficiency and memory change.Although the main sample analysis showed that sleep efficiency was not strongly related to memory decline, we ran a mediation analysis to test for hippocampal MD change as a mediator between sleep efficiency and memory decline , D. Herer = −.04, p = .619, r = −.13, p = .087, APOE, a total of 70 participants carried one or two ε4 alleles. APOE ε4 status was not related to sleep efficiency , or hippocampal MD change . Re-running the main analysis above adding the sleep efficiency PGS and APOE ε4 status, PSQI sleep efficiency still related to hippocampal MD change (p = .031).The sleep efficiency PGS did not relate to worse self-reported efficiency .Higher genetic risk for AD was not related to worse sleep efficiency, that is, higher PSQI scores . We found a stronger relation between sleep efficiency and hippocampal MD change in participants classified as Aβ positive . Comparing Aβ negative and Aβ positive participants did not show differences in sleep efficiency (p = .729), hippocampal MD decline (p = .932), or memory decline (p = .680). When repeating the analysis in the Aβ positive and negative groups separately, we observed a relation between sleep efficiency and hippocampal MD change only in the Aβ positive (p = .019), but not in the Aβ negative subgroup .Of 108 participants with PET data, 23 participants were classified as Aβ positive and 85 Aβ negative , with noSD) age = 69.3 (8.2) years, min−max 44–81 years) had both Aβ and genotype data, and 24 of these participants had one or two APOE ε4 alleles. We included the APOE ε4 status and the PGS for sleep efficiency and AD, respectively, to the initial model. The results demonstrated (1) that sleep efficiency still related to hippocampal MD change differently for Aβ negative and positive participants , with higher propensity of efficient sleep showing less MD hippocampal decline. The AD PGS showed a very weak effect (p = .087), while the APOE genotype showed no effect (p = .537).A subsample of 76 participants (mean (p = .004), and (2) when excluding the participants with depression and MMS scores beyond threshold values, the results remained highly similar. That is, sleep efficiency related to hippocampal MD change (p = .011), as in the main analysis, and weakly to memory change , while the relation between sleep efficiency and hippocampal MD change differed depending on cortical Aβ accumulation (p = .012).We verified that (1) when controlling for levels of depression (both baseline and change scores), sleep efficiency still related to MD hippocampal change   and 66  participThe mechanisms of microstructural hippocampal decline remain unclear, but may relate to decay of the dendritic architecture. In mice, hippocampal dendritic spine densities have been shown to be reduced in aging , and aftThe relation between sleep and Aβ appears reciprocal, as Aβ accumulate after sleep deprivation , and incAs we observed a relation between sleep efficiency and memory decline mediated by higher hippocampal diffusivity, we hypothesize that hippocampal decline, when concomitant with cortical Aβ accumulation, causes sleep problems, here in the form of poorer sleep efficiency, and memory decay. A candidate mechanism could be subtle alterations of sleep-related cortico-hippocampal coupling. For instance, during sleep, slow waves  propagate from the cortex to hippocampus , and bilThe current data does not allow inferences that rule out the reverse causality, of sleep affecting hippocampal decline, potentially partly via reduced glymphatic clearance of potentially neurotoxic substrates which might predispose to Aß and tau pathology  However,APOE ε4 allele. The latter finding was in agreement with previous findings [APOE and the PGSs, we observed weak relations with sleep efficiency and hippocampal diffusivity change, respectively. Such associations must be resolved in larger samples before we can draw the conclusion that the relation between sleep efficiency and hippocampal decline is partly independent of genetics.The associations remained after controlling for genetics risk indexed by PGSs for sleep efficiency and AD, respectively, as well as the presence of the findings , 24. Forfindings . No relafindings . For botLimitations of this study include the use of a self-report measure of sleep, at one time point, not necessarily in the same month as baseline MRI, instead of objective measures such as activity monitors, or polysomnography, collected repeatedly, starting within a month of the baseline MRI. Although self-reported sleep measures might provide more representative data on sleep than a single-night polysomnography , a relatThe results indicate that hippocampal microstructural decline related to sleep efficiency in Aβ positive participants, and mediated the link between sleep and episodic memory change across the adult lifespan. This relation was not readily explained by genetic effects. Poor self-reported sleep efficiency might constitute a separate risk marker for AD, and future studies need to address why sleep is related to more hippocampal decline in Aβ positive older adults even without dementia.zsab110_suppl_Supplementary_MaterialsClick here for additional data file.


Values of δ34S were significantly lower in hair samples from inland regions relative to those from coastal locations, and a similar pattern was observed in δ15N values. Understanding geographic variation in δ34S and δ15N values in human hair is useful for provenancing humans in South Korea.Dietary homogenization has progressed worldwide due to westernization and the globalization of food production systems. We investigated dietary heterogeneity in South Korea by examining the spatial distribution of carbon (C), nitrogen (N), and sulfur (S) isotope ratios using 264 human hair samples. Overall, variation in isotope values was small, indicating low dietary heterogeneity. We detected differences in  Stable isotope analyses of bio-elements, including carbon (C), hydrogen (H), nitrogen (N), oxygen (O), and sulfur (S), are commonly used to investigate the dietary protein sources consumed by both contemporary and past human populations –4. DietaOver the past two decades, scientists have investigated the spatial distribution of bio-element stable isotopes in human tissues for forensic analyses , 10. Amoδ13C and δ15N values among countries or regions. Broadly, human dietary patterns have homogenized due to rapid globalization in food production and distribution processes, resulting in reduced differences in isotope values among regions [Unlike H and O isotopes, the stable isotope ratios of C, N, and S are typically used for dietary reconstruction studies because their ratios in human tissues are entirely related to food consumption . Therefo regions , North A regions , 19, andδ13C values in human hair is strongly related to the amount of C4 plants in human diets [δ15N values are reflective of trophic level, and can thus indicate whether an individual is omnivorous or vegetarian if they consume only terrestrial foods [δ15N values than those who mainly eat terrestrial foods [δ34S values are reflective of the consumption of foods from different geological regions. Specifically, marine-derived foods exhibit approximately +20 ‰ of δ34S values [δ34S values can reflect both diet and geographic origin [δ34S values include bedrock geochemistry [δ34S values in the USA using human hair samples, where inland values were much lower than those from coastal regions. These regional differences can then be applied in forensic provenancing studies [The global spatial distribution of an diets , 20 and al foods . Generalc origin , 22, 23.hemistry , 22, atmhemistry , 24, and studies , 19.δ13C and δ15N values in human hair available from South Korea. To our knowledge, no study has examined spatial variation and heterogeneity in C, N, and S isotopes in human hair at a national scale in South Korea.Datasets of stable isotope values determined from human hair are highly biased to North America and Europe, with limited data available from other countries . Minimalδ13C, δ15N, and δ34S values in human hair samples to extend the applicability of isotopes for human provenancing using a multi-isotope approach. We further investigated dietary heterogeneity and characterized the dietary patterns of contemporary Koreans using C, N, and S isotopes. Finally, we compared isotope ratios from the Korean hair samples with those from other countries to characterize dietary protein sources in Korea in a global context.The use of multi-isotopes from human tissues improves the predictive power of isoscapes . BuildinThis study was approved by the Bioethics Committee at the Korea Basic Science Institute (IRB number: 005). Hair samples were collected from trash bins with oral consent of the owners of barbershops. Only samples identified as male hair were collected from barbershops. Biological sex was the only donor information collected. Isotope data and sample locations are available in We collected 264 human scalp hair samples from trash bins at barbershops for men in South Korea. Hair samples were placed in individual plastic bags at the time of collection. To ensure that the isotopic signature of hair samples reflected the geographic location of sampling regions, hair samples were obtained from barbershops located in residential areas. We assumed that the collected hair samples were representative of men residing in the area around the collection site.Prior to analysis, to remove lipids and surface contaminants, approximately 3 g of hair from a bundle of multiple hairs from the same individual were placed in a 100-mL beaker, washed using 0.5% Triton-X , Milli-Q water (> 18 MΩ cm), and acetone , and then rinsed twice with Milli-Q water. The amount of solvent used for each wash was sufficient to completely submerge the hair samples, and each step was treated in an ultrasonic bath for 2 min. Washed samples were then fully dried in a drying oven at 60°C overnight. Each dried sample was pulverized into a fine powder using a ball mill . Homogenized samples were stored in vials prior to analysis. For the isotope analysis, 34 food samples were collected from supermarkets in metropolitan cities, subsequently, lipids were removed and samples were freeze-dried at −80°C for 72 h and then ground using a ceramic ball mill.2 gas; the analyzer was operated in continuous flow mode. For the C isotope ratio, approximately 0.1 mg of prepared hair sample was encapsulated in a tin container . For the N and S isotope ratios, approximately 0.8 mg of prepared hair sample was encapsulated in tin capsules, respectively. The tin capsules were stored in a desiccator until analysis to avoid vapor absorption. The encapsulated sample was combusted at 1150°C in a combustion reactor and excess oxides were removed in a reduction reactor at 850°C. Thereafter, N2, CO2, and SO2 gases that had been generated were separated using chromatography and the isotope ratios were measured using the IRMS. Because of varying content of C, N, and S in hair samples, C, N, and S isotopes were analyzed separately.C, N, and S isotope ratios were determined using an isotope ratio mass spectrometer  that was equipped with a vario PyroCube elemental analyzer  that included a combustion tube maintained at 1150°C and a U-shaped adsorption column to separate the generated COδ) relative to an international standard unit per mill (‰) as follows: δ (‰) = (Rsample/Rreference − 1) × 1000, where Rsample and Rreference are the molar ratios of the heavy to light isotopes of the sample and standard, respectively, representing 13C/12C, 15N/14N, or 34S/32S. The resulting δ13C, δ15N, and δ34S values were reported against Vienna Pee Dee Belemnite (VPDB), atmospheric nitrogen (Air), and Vienna Canyon Diablo Troilite (VCDT), respectively. The analytical precision was within ± 0.1 ‰ for C, ± 0.2 ‰ for N, and ± 0.2 ‰ for S.Stable isotope ratios were reported using the standard delta notation (δ13CVPDB = −29.8 ‰), IAEA-CH-3 , IAEA-CH-6 , IAEA-600 , USGS-40 , USGS-42 , USGS-43 , IAEA-N-2 , IAEA-SO-5 , and NBS-127 .The following standard reference materials were used for the calibration of C, N, and S isotope ratios: NBS-22 .Normality was tested using the Shapiro-Wilk test. P-values were considered significant at the α = 0.05 level. We used Student’s t-tests to compare mean isotope values between two groups and used analysis of variance (ANOVA) to compare isotope values among provinces. Statistical significance was interpreted based on δ13C, δ15N, and δ34S values obtained from the hair samples were visualized across South Korea (including the province of Jeju Island) using interpolation mapping. Maps were produced using ordinary kriging (OK), a common geostatistical interpolation method, and we did not consider the application of other interpolation methods . The OK was applied using ArcGIS version 10.8 . In addition, spatial autocorrelation (Moran’s index) was performed using the software to examine the spatial distribution pattern of human hair .The spatial distributions of the The range and mean values of C, N, and S isotope ratios obtained from hair samples collected from nine administrative provinces and seven metropolitan cities in South Korea are summarized in δ13C value among all hair samples was −19.2 ± 0.5 ‰ (range: −20.9 ‰ to −17.6 ‰). The mean δ15N value was +9.7 ± 0.5 ‰ (range: +8.4 to +11.7 ‰). The mean δ34S value was +7.3 ± 0.8 ‰ (range: +3.4 to +10.0 ‰). The δ13C, δ15N, and δ34S values obtained from food from local stores and supermarkets are shown in Overall, the mean δ13C values in food samples ranged from −29.1 ‰ to −9.9 ‰ (n = 545), indicative of both C3 and C4 plants. The majority of the food samples were reflective of C3 plants , with the exception of one sample greatly enriched in 15N (+23.0 ‰), and approximately 60% of the food samples occurred within the range of +2 to +8 ‰. Thus, the average δ15N value in human hair samples was +5.4 ‰ higher than that from food samples. An elevated δ15N value in hair is strongly related to animal protein consumption and is reflective of individual dietary patterns. The supply of animal-based protein  is significantly higher than plant-based protein  in South Korea [δ15N values in Korean hair samples.Unlike  samples , where tth Korea . Thus, aδ34S values from both food and hair samples showed a similar pattern to that of δ15N values, and seafood samples with relatively higher δ34S values were more clearly separated from human hair and other food samples  and garlic [δ15N and δ34S values between coastal and inland crops. Such spatial distribution may indicate little or no sea-spray effects around coastal areas in the mainland. However, Jeju Island (JJ), located in the southern part of the Korean Peninsula is the administrative province most affected by sea-spray. Jeju Island samples were collected close to the coast, so local foods may have contained particularly high amounts of marine aerosols. Garlic samples collected from Jeju Island had slightly higher δ34S values than those collected from the mainland, indicating a weak sea-spray effect. Meanwhile, δ15N values of pork meat samples produced on Jeju Island did not differ from those produced on the mainland  [δ34S data for animal meats such as pork, beef and chicken are currently available for South Korea. Therefore, given the δ15N and δ34S data of agro-products available for South Korea, high values of δ34S and δ15N in coastal human hairs from the mainland can primarily be interpreted as a result of fish and seafood consumption [The  samples . General in hair , and higd garlic  samples  n = 16) . No δ34Ssumption .δ13C and δ15N values from food samples were scattered across a wide range, whereas those from hair samples were confined to a narrow range. This may be because hair reflects the average isotope values of various food samples.The results of the isotope analyses of hair and food samples are shown in δ13C values  and tofu made from soybean (C3 plant) had low C values ; δ13C val manure . In Souttivation . The higtivation . In addiδ13C values, whereas cattle in the USA and Mexico are typically fed corn, resulting in high δ13C values. In Korea, C3 and C4 plants are used to feed cattle; thus, δ13C values are intermediate to beef from the other continents [δ15N value that was approximately +3 ‰ higher than that of pork or chicken . Comparisons of isotope values for all provinces and cities are provided in We found no significant difference in the intake rate of animal and marine foods between metropolitan cities and administrative provinces in South Korea. On average, animal and marine food intake is reportedly only 1.2% higher in metropolitan cities in South Korea . Data rerovinces  and smalδ13C values were slightly higher in Jeju Island samples (−18.9 ± 0.4 ‰) and lowest in Jeonbuk (−19.6 ± 0.6 ‰). Mean δ15N was highest in Jeonnam (+10.0 ± 0.6 ‰) and lowest in Chungbuk (+9.3 ± 0.3 ‰), the only inland province that is not in contact with the sea. The mean δ34S value was highest in Jeju Island (+7.8 ± 0.7 ‰) and lowest in Chungbuk (+6.8 ± 0.3 ‰). Intra-region variation in δ34S was highest in Gangwon (± 1.2 ‰). Overall, the variation in δ34S values was low and did not deviate significantly from the mean.Among all provinces, mean Dependency on imported food, rather than food sovereignty, is increasing annually in South Korea , which wTo characterize the dietary patterns of contemporary Koreans on a global scale, we compared the isotope ratios obtained from the hair samples with those from other countries . We focuδ13C and δ15N values in Korean hair obtained in this study are similar to those reported previously [δ13C and δ15N values in Korean hair were similar to those in Japanese hair [δ13C values, the consumption rates of C3 and C4 plants are similar between Japan and Korea  [δ13C and δ15N values, male Koreans eat both C3 and C4 plants and are omnivorous.For comparison, the USA, which mainly consumes C4 plants reflected by high C values , and theC values , 19, 44 C values –48 are a Canada) , 50 and  Canada) , 52, sugδ15N values obtained from contemporary Korean and Japanese hair samples should be similar, as was observed here.According to the Food and Agriculture Organization of the United Nations , Korean δ13C and δ15N values were higher in Korean hair relative to Chinese samples . However, data from Mongolia deviated considerably from the correlation line .δ13C and δ15N values in Korean and Japanese hair samples were smaller than those of Chinese hair samples. This is likely because foods consumed by Korean and Japanese people are characteristic of a global supermarket, with more homogenous isotope compositions [The range of variation of ositions , indicatδ13C, δ15N, and δ34S values in human hair samples for South Korea are shown in δ13C values were slightly higher than in other regions. This was a consistent, albeit weak, pattern that was reflected in other metropolitan cities. Jeju Island, which sees high international traffic, also had higher δ13C values. Globalization has had the most rapid advancement in metropolitan cities and on Jeju Island within South Korea. Westernization and globalization tend to relate to increased consumption of foods containing corn [δ13C values [δ13C values in human hair. Excluding the areas mentioned above, we saw little effect of geographic and environmental factors on δ13C values , that is, distinctively low in mountainous areas and high in agricultural areas  are primarily used in agricultural lands in South Korea [δ15N values seems to be related to the consumption of food produced in these areas. Generally, δ15N values are also determined by the consumption of fish and meat [δ15N values [By contrast, there was a distinct geographic pattern in al areas . Organicth Korea , 58; thuand meat , 59, 60 N values , 62.δ34S values of human scalp hair in South Korea showed a distinct geographic pattern similar to that of δ15N values  from human hair samples. However, a stronger correlation was observed between δ15N and δ34S values .Finally, N values , signifiproducts , 24. A wδ15N and δ34S values for human hair. For the δ15N and δ34S values, the Moran’s I index was 0.356  and 0.141 , respectively. Data lacking significant autocorrelations suggest that human hairs in terms of δ15N and δ34S values are not spatially autocorrelated but are instead randomly dispersed. This observation can be explained by the possibility that the isotopic compositions of human hair could reflect both geographical and topographical features. Likewise, agricultural practices more extensively occurred in the lower elevation areas than in mountainous areas. The distribution of lowlands and highlands is not consistent with distance from marine environments. Thus, the isotopic value in each sampling region would be compensated or integrated, statistically resulting in the absence of spatial autocorrelation. From this perspective, the randomly dispersed- spatial distribution would also be attributed to westernization and globalization. Nonetheless, the interpolated map illustrates a distribution pattern that reflects land use and distance from marine environments.Spatial autocorrelation was employed to evaluate the distributions of δ13C, δ15N, and δ34S values are useful for provenancing studies and allow us a more holistic view of geographic patterns following the publication of H and O isotope data.Given the patterns we observed among all isotope values, we conclude that dietary patterns have become homogenized in Korea due to westernization and globalization, irrespective of region, but that dietary heterogeneity still exists in some provinces. Our results indicate that C, N, and S isotopic signatures in human hair are strongly correlated with the food consumed by individuals. These regional differences in mean δ13C, δ15N, and δ34S in hair samples were −19.2 ± 0.5 ‰, +9.7 ± 0.5 ‰, and +7.3 ± 0.8 ‰, respectively. The inter-regional difference in C isotope ratios was < 0.5 ‰, indicating dietary homogeneity due to rapid westernization and globalization. Nevertheless, regional dietary heterogeneity still exists in some areas in South Korea.We collected 264 human scalp hair samples across South Korea to investigate dietary heterogeneity and spatial variation in C, N, and S isotopes at a national scale. To our knowledge, this is the first national-scale study assessing C, N, and S isotope ratios in human hair in South Korea. The mean values of δ34S values in human hair were greatly influenced by proximity to marine environments in South Korea, with higher values in coastal relative to inland regions. Interestingly, the N isoscape showed a similar pattern to that of the S isoscape. The consistency between the two illustrates the potential of isoscapes to act as a proxy for food source in South Korea, and can provide useful data for forensics and migration tracking. Further, understanding C, N, and S isoscapes in South Korea contributes to the growing literature on the identification of residential history using isotopic analyses of human hair.Spatial products like isoscapes are valuable tools because they allow us to visualize spatial heterogeneity in isotopes given environmental parameters, such as distance to sea. We found that S1 Table(XLSX)Click here for additional data file.S2 Table(XLSX)Click here for additional data file.S1 Fig(A) Fish and (B) meat per capita supply for East Asian countries since 1961.(TIF)Click here for additional data file.S2 FigFish and animal contribution to protein supply for East Asian countries since 1961: (A) fish/total proteins, (B) animal/total proteins and (C) /total proteins.(TIF)Click here for additional data file.


Howard DPJ, Gaziano L, Rothwell PM, on behalf of the Oxford Vascular Study. Risk of stroke in relation to degree of asymptomatic carotid stenosis: a population-based cohort study, systematic review, and meta-analysis. Lancet Neurol 2021; 20: 193–202—In the table of this Article, the rate (per 100 person-years) of transient ischaemic attack or stroke should have been 0·66 in the 50–69% stenosis group and 3·51 in the 70–99% stenosis group. And the second paragraph of the Results section should read as follows: “Of the 2178 imaged patients, 469 had 50% or greater stenosis of at least one carotid bifurcation, of whom 243 had 50–99% symptomatic carotid stenosis and 11 had an occlusion, and 207 had 50–99% asymptomatic carotid stenosis and eight had an occlusion.” These corrections have been made to the online version as of March 16, 2021.