Sogatella furcifera (Horváth), with finding heteroplasmy phenomenon confirmed by PCR reaction and Sanger sequencing method. This heteroplasmy was not observed in WBPHs (n = 24) collected from the fields, suggesting that it may be uncommon in fields. We also analyzed single nucleotide polymorphisms, insertion and deletions, and simple sequence repeats among three currently available WBPH mitogenomes of Korea and China, suggesting that identified intraspecific variations could be potential candidates for developing markers to distinguish geographical populations of WBPH including Korean and Chinese. Phylogenetic analysis of 32 mitogenomes of Delphacidae including the three WBPH mitogenomes suggested that Delphacinae seems to be monophyletic and Sogatella species including WBPH are clearly formed as one clade.We completed one mitogenome of white-backed planthopper (WBPH), Sogatella furcifera (Horváth), is one of the major sap-sucking rice pests in East Asia. We have determined a new complete mitochondrial genome of WBPH collected in the Korean peninsula using NGS technology. Its length and GC percentages are 16,613 bp and 23.8%, respectively. We observed one polymorphic site, a non-synonymous change, in the COX3 gene with confirmation heteroplasmy phenomenon within individuals of WBPH by PCR amplification and Sanger sequencing, the first report in this species. In addition, this heteroplasmy was not observed in wild WBPH populations, suggesting that it may be uncommon in fields. We analyzed single nucleotide polymorphisms, insertion, and deletions, and simple sequence repeats among the three WBPH mitogenomes from Korea and China and found diverse intraspecific variations, which could be potential candidates for developing markers to distinguish geographical populations. Phylogenetic analysis of 32 mitogenomes of Delphacidae including the three WBPH mitogenomes suggested that Delphacinae seems to be monophyletic and Sogatella species including WBPH are clearly formed as one clade. In the future, it is expected that complete mitogenomes of individuals of geographically dispersed WBPH populations will be used for further population genetic studies to understand the migration pathway of WBPH.White-backed planthopper (WBPH),  Sogatella furcifera (Horváth), commonly called the white-backed planthopper (WBPH), is an important rice pest species in East Asian rice fields  of National Institute of Agricultural Sciences in Korea. The two wild populations, named as WBPHHD and WBPHBS, were sampled at paddy fields in Hadong-gun  and Boseong-gun , respectively, in the Republic of Korea on 19 July 2021.We extracted genomic DNA from a total of 20 WBPH individuals of the Taean population of WBPHs (named as WBPHTA) using the CTAB-based DNA extraction method  for next-generation sequencing. Each genomic DNA of ten WBPH individuals in WBPHTA and 24 WBPH individuals in two wild WBPH populations was additionally extracted and used to determine nucleotide sequences in a polymorphic site of the http://geis.infoboss.co.kr/; 24 November 2021) which have been utilized in the various genomic studies [Genome sequencing was performed using HiSeqX at Macrogen Inc., Seoul, Korea from extracted DNA of the WBPH sample. De novo assembly and confirmation were done by Velvet v1.2.10  after fi studies ,51,52,53® v2020.2.4  was used to annotate mitochondrial genome based on the previously sequenced mitogenome of WBPH (NC_021417) [Geneious Prime_021417)  with ARW_021417)  for anno4667) in the whole WBPH mitogenome, we designed a PCR primer set, Sofur-mt4218F (5′-ACA CTA ACC TAA TAT TTG CC-3′) and Sofur-mt4945R (5′-GAT GCT CCT GAT CTT AAT AA-3′). PCR amplification of each genomic DNA for ten WBPH individuals of WBPHTA, eight individuals of WBPHHD, and 16 individuals of WBPHBS was conducted with PrimeSTAR GXL DNA polymerase  by following 30 cycles of 10 s at 98 °C, 15 s at 50 °C, and 1 min at 68 °C. Sager sequencing of PCR products was carried out by the DNA sequencing service of Macrogen Inc. Korea using ABI 3730xl System .To verify the sequence of one polymorphic site  [To predict the three-dimensional structure of COX3, Swiss-Model (er 2021)  was used® v2020.2.4 , which has been utilized in the previous insect mitochondrial genome studies [Pair-wise sequence alignments among complete mitogenome of WBPH including the mitogenome sequenced in this study were conducted by MAFFT v7.450  with the studies ,62,63,64http://ssrdb.infoboss.co.kr/; 24 November 2021) which has been utilized in the previous studies [Simple sequence repeats (SSRs) were identified on the mitogenome sequence using the pipeline of the SSR database  to identify intraspecific variations of SSRs. The pipeline of the SSR comparison implemented in the SSRDB used in various organelle genome studies [−10 and a maximum flanking sequence for the comparison of 60 bp.SSRs identified from the three WBPH mitogenomes were compared based on their flanking sequences under the environment of the SSRDB  in the Cixiidae family [®2020.2.4 . The HKY85 model with gamma rates was used as a molecular model. A Markov chain Monte Carlo algorithm was employed for 1,100,000 generations, sampling trees every 200 generations, with four chains running simultaneously. Trees from the first 100,000 generations were discarded as burn-in.The 33 mitogenomes including 32 representative mitogenomes (30 species) selected for each species in the Delphacidae family and one outgroup mitogenome of e family  were alie family , and alie family . In the e family  plug-in S. furcifera  with a length of 16,613 bp, containing 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), and 2 ribosomal RNAs (rRNAs)  , which i (rRNAs) . The GC  (rRNAs) .The AT-rich region, considered as a putative control region of the WBPHTA mitogenome, is 2219 bp long, which is the shortest length among the three WBPH mitogenomes . The varNilaparvata lugens (Stål)) [L. striatellus) [The length variations of complete mitogenomes of two planthopper species, the brown planthopper ) ,76,77,78atellus) ,77,79, watellus) ,76,77,78atellus) ,77,79, wCOX3 at 4667th starting from tRNA-Ile on the mitogenome, presented A and T together  by PCR and sequenced the PCR products using the Sanger sequencing method. We obtained the sequencing results from nine out of ten individuals B, resultCOX3 from a total of 24 individuals of the two Korean wild WBPH populations, WBPHHD and WBPHBS, was investigated using PCR and the Sanger sequencing method. In the results, there was no heteroplasmy phenomenon and single nucleotide polymorphism  at the 4667th of the WBPH mitogenome. It suggests that the heteroplasmy and SNP of WBPHTA mitogenome might be uncommon in the wild WBPH populations in fields. The heteroplasmy of WBPHTA mitogenome would be associated with continuous pressures from the inbreeding environment in the laboratory. It is remained to study whether this heteroplasmy phenomenon of WBPH is artificially created through continuous insect rearing indoors.Because this heteroplasmy phenomenon was only observed in WBPH individuals from WBPHTA, a breeding line, has been established and reared in the insectary since 2006, it was necessary to evaluate whether this heteroplasmy in the WBPH mitogenome is also common in wild populations or not. A 4667th nucleotide sequence of COX3 was deduced by changing A to T at the 4667th position. Moreover, there was one HexaSSR, H0000007  to phenylalanine (F) in H0000007 , which cSS-MODEL  based on helixes , which iCimex lectularius Linnaeus, Hemiptera:Cimicidae) [Apis mellifera Linnaeus, Hymenoptera:Apidae) [Ectatomma ruidum (Roger), Formicidae:Ectatomminae) [Anapodisma miramae Dovnar-Zapolskij (Orthoptera:Acrididae) [Tetrodontophora bielanensis (Waga) (Poduromorpha:Onychiuridae) [Drosophila melanogaster Meigen (Diptera:Drosophilidae) and is caused by paternal mitochondrial DNA leakage [The heteroplasmy in the mitogenome has been found in various insect and mite species, including the bed bug (micidae) , honeybe:Apidae) , a neotromminae) , Anapodirididae) , Tetrodoiuridae) , and Dro leakage ,86. It h leakage ,88,89,90 leakage . Taken tBased on three pair-wise alignments of WBPH mitogenomes, 21 SNPs and four insertions and deletion (INDEL) regions  and six SNPs and five INDEL regions  against WBPHHN and WBPHYN mitogenomes were identified A. The loND2, COX1, COX3, ND3, ND5, ND4, CytB, and ND1, contained one or more SNPs. Four of eight PCGs, COX3, ND5, ND4, and CytB, contained one non-synonymous SNP in each pair of WBPH mitogenomes. These non-synonymous SNPs can affect the function of the PCGs as reported for L. striatellus [Thirteen of 21 SNPs (61.90%) between the WBPHTA and WBPHHN mitogenomes, six of six SNPs (100.00%) between the WBPHTA and WBPHYN mitogenomes, and 11 of 14 SNPs (78.57%) between the WBPHHN and WBPHYN mitogenomes were found in PCGs. Eight of 13 PCGs, including iatellus . These PL. striatellus [N. lugens [Chilo supressalis  [Aphis gossypii Glover [Spodoptera frugiperda (J. E. Smith) [Hipparchia autonoe (Esper) [Alphitobius diaperinus Panzer [The numbers of SNPs and INDEL regions identified from the three WBPH mitogenomes were relatively smaller than those of iatellus ,79, N. l. lugens , and Chi , of whici Glover ,94,95 an. Smith)  but were (Esper)  and Alphs Panzer .Stegobium paniceum   and Monomorium pharaonis  .Simple sequence repeats (SSRs), which can be utilized as molecular markers to distinguish insect species ,98 and tCTYB gene  the putative control region  but was lower than that of S. paniceum .Comparisons of flanking sequences of each SSR identified from the three WBPH mitogenomes using the SSRDB , identified 108 SSR groups: 98 out of 108 SSR groups (90.74%) contained three SSRs originating from three WBPH mitogenomes, called common SSRs B. Three l region . Interesl region B, indicaND4 and ND1 genic regions. Interestingly, the 5′ part of the putative control region displayed a high density of SSRs in a conserved manner across the three mitogenomes, while the 3′ part of the putative control regions exhibited a low density of SSRs with variable manners across the three mitogenomes). This correlation between SSR density and intraspecific variations will be investigated more in other insect species, which can be used as a guide for developing efficient molecular markers using SSRs. The intraspecific SSRs identified in this study could be potential candidates for developing markers with WBPH mitogenomes which will be sequenced in near future to distinguish geographical populations and to understand the migration pathway of WBPH.The SSRs identified from the three WBPH mitogenomes were densely distributed in the putative control regions as well as in the Haplaxius crudus van Duzee, 1907, were constructed. Among the 32 Delphacidae mitogenomes, only one mitogenome, Stenocranus matsumurai Metcalf, 1943(GenBank accession number: MH293469), belongs to the subfamily Stenocraninae, while the remaining 31 mitogenomes belong to the subfamily Delphacinae. Phylogenetic trees demonstrated that both subfamilies were clearly distinct from each other  and Bayesian inference (BI) phylogenetic trees of 32 mitogenomes of Delphacidae including three WBPH mitogenomes and one outgroup species, ch other . In addich other  were supch other , displaych other  was alsoch other , indicatCOI sequence data [The BI tree demonstrated that three WBPH mitogenomes were clustered in one clade; while the ML tree demonstrated that WBPHTA and WBPHHN were clustered together, suggesting the possibility that the WBPHTA sample may be from a similar region to the Hainan province in China, where the WBPHHN isolate was captured , togethence data . FurtherCOX3, causing not severe conformational change based on the prediction of three-dimensional structure. In addition, this heteroplasmy was not found in the two wild populations in Korea, suggesting that continuous selection pressure may cause this heteroplasmy. Intraspecific variations and simple sequence repeats (SSRs) were identified among the three WBPH mitogenomes, suggesting that these can be a potential target to develop molecular markers to distinguish populations of WBPHs from different locations. Phylogenetic analysis suggested that Delphacinae is monophyletic and WBPH, as one group, is clearly divided with closely related Sogatella species. The heteroplasmy, the firstly reported in WBPH mitogenome, suggests an additional step to check the polymorphic sites during the assembly. In addition, intraspecific variations in WBPH mitogenomes can be useful to understand its genetic diversity as well as to develop useful markers and to understand the migration of geographically dispersed WBPH populations.In this study, we reported a new complete mitogenome of WBPH collected in Korea (named as WBPHTA) and firstly identified heteroplasmy in WBPH and Delphacidae. The heteroplasmy resulted in the amino acid substitution in


Since 2010, States party to the United Nations Framework Convention on Climate Change have recognized planned relocation as a viable adaptation to climate change. Planned relocation has been attempted in many communities globally and has raised serious issues of equity in some cases. Implementation driven by principles of equity is crucial in ensuring successful planned relocations that decrease loss and damage. In this Policy Analysis, we put forth a framework for equitable planned relocation rooted in theories of justice as a basis for implementation. The framework centers around three principles: comprehensive recognition of affected stakeholders in decision-making, consideration of socio-cultural risk factors relevant to relocation, and evaluation of multiple measures of well-being. There are many actors involved in planned relocation. Unique features and abilities of international organizations lend themselves to promoting equitable planned relocation in partnership with other stakeholders. Through the exploration of case studies, we identify best practices that international organizations have available to influence the design, implementation, and evaluation of planned relocation processes. These practices are relevant when striving for equity for all affected individuals and communities. Points of intervention include agenda-setting and advocacy, funding and implementation standards, and facilitation of international cooperation. International organizations also face barriers to supporting equitable planned relocation. Limitations include lack of enforcement mechanisms, limited resources, and fundamental dependence on existing governance structures and global collaboration. As the necessity of planned relocations grows, the need for leadership from international organizations in implementation is magnified, underscoring the importance of developing and evaluating approaches to just implementation. It is affecting resources such as potable water and fertile land, rendering some territory uninhabitable as a result of sea level rise, and severely impacting infrastructure due to extreme weather events  have recognized mobility via planned relocation as a viable adaptation to climate change. Planned relocation is a response to environmental and climate hazards in which people move, on their own or with assistance, to a new location with the ability to rebuild their lives (UNHCR 2016). Although we focus on environmentally driven relocations, it is important to note that relocations can be driven by a host of issues, including development, conflict, and politics. We will discuss a few examples, especially related to development, where relevant for climate and environmental relocations. Managed retreat is a subset of planned relocation that focuses on mitigating the impacts of sea level rise and other hazards and then returning the land to open space  and national-level agencies that work in the planned relocation space are also germane.International organizations have exhibited commitment, leadership, and action around climate adaptation and risk reduction  in 1979 established principles of equality and has led to the striking down of many laws that upheld the discrimination of women  has a policy that provides special consideration regarding the risks of impoverishment associated with relocation when working with low-income groups IDB . The WorAs such, international organizations and their partners have made great strides towards equitable adaptation thus far. Nevertheless, the problems that the global community faces in this realm are enduring and require sustained action. Importantly, the massive moment of disruption the globe is facing as a result of the COVID-19 pandemic has the potential to threaten this progress and further undermine equity. It is therefore critical that international organizations leverage their unique features and abilities to promote equitable planned relocation.Out of many features and abilities held by international organizations, we highlight three that are most relevant for promoting planned equitable relocation. Table International organizations play a key role in setting the international agenda, which can guide the attention of governments, NGOs, and other international organizations towards coordinated action. They can identify gaps and advocate for an equitable approach to planned relocation. The reach of international organizations in agenda-setting extends from applications such as implementing planned relocation on the ground, redefining sovereignty for nations who have lost their territory, and abiding by principles of justice through the centering of Indigenous peoples.International organizations have had many successes in agenda-setting. A key example is that of gender mainstreaming, which came out of the Beijing Platform for Action at the 1995 UN Conference on Women and led to the prioritization of gender equality within the UN, its member states, and the broader NGO and philanthropy field . It is therefore critical that a shared understanding of the possible roles of each stakeholder and implications for the whole system be created . It was not only facilitating the day-to-day activities but also working with the government of Bangladesh to strategically plan for the future of the camps and those in them  facilitated by the International Organization for Migration (IOM). The PIDC includes 21 states and territories that seek to develop policy around migration flows. Together, they have sought to address immigration processes within their borders in the wake of a natural disaster by specifying the role of immigration officers, delineating the separation of individual documentation and disaster relief, and prioritizing proactive planning (PIDC 2010). The PIDC includes member states and territories that might otherwise be left out of the conversation on migration policy in the region yet are largely affected by it. In coordinating the convening of the PIDC, the IOM fills a unique role in employing the principle of recognition at the governmental level.There are many barriers that international organizations face in implementing planned relocation, highlighting the importance of partnership with other stakeholders. The limitations we delineate are not necessarily unique to international organizations but remain particularly challenging due to the scale at which they impact international organizations.The efficacy and power of international organizations are dependent on geopolitics and international relations broadly Gabriela . As suchInternational organizations take on many projects and priorities that have the potential to conflict with the long-term planning of relocation Graham . For insPlanned relocation is a resource-intensive process. Some international organizations can fund projects as intensive as this, such as the World Bank. Many other international organizations do not have the funding to do so, making them reliant on a patchwork of external funding sources, which could further introduce conflicting priorities Kerlin .Without sufficient resources, applying the framework for equitable planned relocation becomes almost impossible. In order to recognize community members in the planning and decision-making process, many person-hours are needed to build and maintain relationships. Following this process, the creation of multiple options that take into account the various socio-cultural risk factors of the community members requires funding to organize and implement. Lastly, the longitudinal evaluation of multiple measures of well-being and the ability to act on those outcomes requires more person-hours and investment dollars than a typical project.As the hazards posed by climate change continue to rise in frequency and severity, planned relocation is an increasingly important adaptation mechanism. Especially in the current moment, the intersection of climate hazards, public health disasters, and border restrictions creates an urgent need for proactive action with a deliberate framing around equity in order to prevent the occurrence of compounded injustices. To this end, actors in the space should incorporate the framework for equitable planned relocation that we have proposed which encompasses comprehensive recognition of affected stakeholders in decision-making, consideration of socio-cultural risk factors relevant to relocation, and evaluation of multiple measures of well-being into their planned relocation efforts. International organizations have features that uniquely position them to implement the aforementioned principles. These include their abilities to agenda-set and fund, disseminate best practices and promote accountability, and facilitate coordination among relevant actors. It is critical to the success of equitable planned relocation efforts that international organizations do not act alone, but rather leverage their unique points of intervention in partnership with other governmental and non-governmental actors. Future areas of action include implementing, evaluating, and iterating upon the framework for equitable planned relocation.


Variable temperature (VT) 1H NMR measurement, from 316 K to 340 K, was conducted on a suspension of potato starch and deuterium water as well as a mixture of potato starch, NaCl and deuterium water. The hydration level of starch was determined with the increase of temperature. In the presence of NaCl, the initial gelatinization temperature of potato starch was decreased from 331 to 328 K. Meanwhile, in situ1H NMR spectroscopy as a function of time was also carried out to monitor the gelatinization with a time resolution of 90 s per spectrum. Furthermore, the effect of using different processing methods during gelatinization, including varying the temperature or time duration, was investigated in detail. It was confirmed that protons from different groups of starch showed different accessibility for water during hydration of starch granules. In comparison with temperature, gelatinization time as the major factor for reaching complete gelatinization was confirmed. We expect that this research, as a continuing effort to apply NMR spectroscopy for characterizing starch, will pave a new way in the structural elucidation of starch.The gelatinization of potato starch and the effect of NaCl on starch gelatinization were monitored successfully  in situ by 1H NMR spectroscopy.The gelatinization of potato starch and the effect of NaCl on starch gelatinization were monitored successfully  It is commonly considered that native starch is a mixture, which consists of linear amylose and highly branched amylopectin.7–12 The difference in amounts and ratio of these two polymers organized within starch granules, gives rise to considerable variability in properties, such as swelling, pasting and gelatinization behaviours.13–16 Gelatinization, when starch was heated in water, is one of the most significant processes in the industrial application and development of starch.17–19 Under this condition, starch granules undergo disruption of crystallinity and molecular organization. When heating starch in excess water is continued, the granules will suffer an irreversible phase transition, commonly referred to gelatinization.20–25 It is generally considered that the functional properties of starch are mainly determined by the extents and effects of gelatinization treatments. Besides, the functionality of starch is intimately related to its hydration levels. A detailed understanding of the hydration of starch will be a significant asset in developing new starch materials with well-designed functionalities.25,26 Besides, adding salts is a common process to make the starchy derivatives as normal. It has also been proved that salts such as sodium chloride (NaCl) had a significant effect on the gelatinization of starch.27 The effect of sodium chloride could affect the temperature and enthalpy of gelatinization, and simultaneously change the rate and degree of gelatinization.28,29 Therefore, it is of importance to understand the detailed gelatinization process where starch is subjected to sodium chloride solution. However, the gelatinization mechanisms in the presence of sodium chloride are still not well understood. In order to manufacture the renewable biomaterials and obtain high quality of foods, it is necessary to get an improved understanding of starch gelatinization and the effect of salts on gelatinization at the molecular level.As one of the most abundant and attractive natural biopolymers, starch is the main constituent used to manipulate the quality of food in the human diet, and can be widely found in various botanical sources such as potato, corn, wheat and so on.29–37 However, most of these methods were carried out depending on the post treatment, which usually processed samples with several preparation steps. This will result in the detailed process of gelatinization cannot be monitored carefully in real-time. Generally, it was considered these ex situ characterization methods were challenging, due to high sensitivity to sample preparation and limited lifetimes of crystalline structural disruption during gelatinization. Fortunately, the in situ measurements, possessing the powerful performance in real-time monitoring could make up this shortage.18,19,38–43 For example, rapid visco analysis (RVA), monitoring the process in situ, has been widely applied to investigate the gelatinization of starch.38 Recently, waxy and high-amylose maize starch gelatinization process was monitored in situ via synchrotron XRD in combination with a diamond anvil cell at very high pressures.41 Meanwhile, the gelatinization processes of starches from a variety of plant sources were in situ observed by a method of polarizing microscope in combination with a hot stage.42 All of these techniques studied the process via the difference in macroscopic property during starch gelatinization. The detailed structural characterization during the hydration of starch on atomic level remains to be fully studied. However, to the best of our knowledge, there is still few studies employed in situ techniques at atomic-levels to monitor the process of starch gelatinization so far. Variable temperature (VT) NMR spectroscopy, which has been widely applied for investigation of structural and phase transition of materials at atomic-levels, undoubtedly is the suitable technique of choice for probing the gelatinization of starch granules.Considerable effort has been devoted to monitor the process and discovered the mechanism of gelatinization, using a variety of microscopic, diffraction and spectroscopic techniques, including scanning electron microscope (SEM), powder X-ray diffraction (XRD), differential scanning calorimetry (DSC), Fourier transform infrared (FT-IR) and NMR spectroscopy.in situ method for monitoring the detailed process and structural changes that occur in chemical or non-chemical transition.44,45 Compared with other characterization measurements, NMR could acquire the information of structural transition while heating the samples, which makes it possible to probe the transformation process only taken place in high temperature. It was known that starch gelatinization was such a process occurred above gelatinization temperature. Besides, in comparison with the studies of gelatinization using other techniques,46 NMR spectroscopy could achieve the detailed information of phase transition process at molecular level.47 And on this basis, in situ NMR spectroscopy strengthens characterization technique, which makes it possible to real time detect the evolution of hydrogen bonds during gelatinization. Therefore, we envisioned that VT NMR spectroscopy may work as a powerful analytical technique to real time observe the gelatinization process in situ.NMR spectroscopy is a non-invasive and in situ VT 1H NMR spectroscopy to real time monitor the gelatinization process of potato starch. VT 1H NMR measurement, from 316 K to 340 K, was carried out on the suspensions of potato starch and deuterium water with the weight ratio of 1 : 7.5. The details of gelatinization were monitored under the treatment of increasing temperature. The hydrogen bonding interaction between the hydroxyl and hydrogen atoms of starch and water was formed gradually with the increase of temperature. Meanwhile, in situ1H NMR spectra as a function of time (from 0 min to 81 min) were also carried out to monitor the detailed structural transition at atomic level during gelatinization at 328 K. We found that the peak intensity of α-1,6 glucosidic bonds increased obviously with the extending of heating time gradually. Furthermore, the effect with different processing methods during gelatinization, including varying temperature or heating time, was investigated in details. Under different treatment, it was verified amylose and amylopectin showed the different accessibility for water during gelatinization. It was also found that the addition of NaCl decreased the initial temperature of starch gelatinization.In this study, we expected to extend the research utilizing 2.2.1.Potato starch was purchased from Shanghai Yuanye Biological Reagent Co., Ltd. Unless otherwise stated, all other reagents were of analytical grade.2.2.1H NMR spectra were recorded in D2O solution at the frequency of 400.06 MHz on a Bruker Avance III 400 MHz NMR spectrometer. Potato starch powder was suspended with D2O in a weight ratio of 1 : 7.5. Samples were prepared by weighing ca. 80 mg of starch into a NMR tube, then ca. 600 mg of D2O were added to reach the exact ratio of 1 : 7.5. The starch/sodium chloride solution systems were prepared by weighing ca. 80 mg of starch, 25 mg NaCl, and 600 mg of D2O into a NMR tube. The variable-temperature NMR measurements were carried out in the range of 316–340 K with the same probe using a Bruker temperature controller. The samples were heated at the target temperature in steps and kept equilibration for approximately 10 minutes before data acquisition. The experiments were recorded with the π/2 pulse of 14 μs, the repeated scans of 16, and the recycle delay of 1 s respectively. Each of the 1H NMR measurements takes one and a half minutes without the time for varying temperature. The 1H chemical shifts were referenced to the signal of DSS.Liquid 3.3.1.in situ at atomic-levels, the variable temperature 1H NMR measurements at T = 316–340 K were conducted upon the suspensions of potato starch and deuterium water with the weight ratio of 1 : 7.5  could be observed obviously, which indicated that the gelatinization process did not occur at 316 K. When the temperature was increased to 331 K, a broad signal (centred at 5.3 ppm) was recorded at down-field, due to the 1H homonuclear dipole–dipole interactions. We speculated that this peak may be derived from H-1, which were attributed to both the protons of α-1,4 bond and α-end group in starch, and this result was consistent with the Larsen's work.51 They applied the 1H high-resolution MAS NMR spectroscopy to characterize the signals from α-1,4 bond (5.4 ppm) and α-end group (5.2 ppm) of potato starch, respectively. This suggested that the end groups including α-1,4 glucosidic backbone and α-end group were easily accessible for water instead of other groups in starch. It should be noted that only the protons located in mobile regions are detected via1H NMR. Instead, protons from immobile starch regions do not contribute to 1H NMR spectrum. Hence, the strong peak centred at 5.3 ppm indicated that α-1,4 bond and α-end group were located on the mobile regions. The absence of protons from α-1,6 backbone was detected, which was consistent pioneer's work.52 It has been proved that protons α-1,6 backbone was stabilized by the presence of structural water at a low degree of gelatinization, which strongly increased the dipole–dipole interactions and do not contribute to the 1H NMR spectra. Meanwhile, a new broad signal from the H-2,3,4,5,6 of starch was observed, indicating that the interaction between protons of H-2,3,4,5,6 in starch and water molecules began to occur at 331 K. As the temperature increased, for example at 340 K, the peak width at half height of H-1 was decreased from 90 Hz to 34 Hz, which was attributed to the higher extent of hydration. Besides, it was considered that amylose has the more reducing end groups than amylopectin, due to the lower molecular mass of amylose. Hence, the signal of H-1 also demonstrated that the end groups of amylose showed the better accessibility and higher degree of hydration for water than amylopectin. Furthermore, as the temperature increased to 334 K, two well-resolved peaks of H-2,3,4,5,6 centered at 3.6 and 3.8 ppm were observed, which were from the signals of H-2,4 and H-3,5,6 respectively. This observation suggested that the gelatinization process of potato starch was started at 331 K. When the temperature increased to 337 K, the peak of H-3,5,6 split into two sub-peaks caused by further progress of the gelatinization, and these two sub-peaks become well resolved as temperature increased to 340 K with chemical shifts of 3.8 and 3.9 ppm, which were attributed to the group of H-5,6 and H-3, respectively. It was also indicated that this suspension was hydrated to the large extent of gelatinization. In order to better distinguish these two peaks of H-5,6 and H-3, as displayed in 53 respectively. Meanwhile, the 1H NMR spectrum at 340 K was almost consistent with that of 337 K, indicating that the suspension of potato starch and deuterium water with the weight ratio of 1 : 7.5 might be gelatinized to reach upper limit to the extent of gelatinization.To real-time monitor the detailed gelatinization process of potato starch  1 : 7.5 . Under tin situ monitor the gelatinization of potato starch in the presence of NaCl, the variable temperature 1H NMR at T = 316–340 K were carried out upon the sample of potato starch, NaCl and deuterium water (donated as NaCl@potato starch). It was found that the 1H NMR spectra of NaCl@potato starch were consistent with that of potato starch during the whole variable temperature process. As shown in 1H NMR (seen in 2O.In order to  seen in . This re3.2.in situ1H NMR spectra as a function of time (from 0 min to 81 min) were carried out to monitor the detailed structural transition at atomic level during gelatinization as shown in 1H NMR signals of starch cannot be resolved unambiguously, which indicated the starch hydration has not occurred yet. After the first measurement, 1H NMR signals of starch protons could be detected obviously, which was attributed a small amount of starch begins to dissolve in deuterium water and the gelatinization occurred. As the increase of the experimental time, the signal intensity of starch was increased, due to the well solubility of starch at high temperature. This indicated starch gelatinization happened within a large extent. At experimental time of 45 min, the well resolved 1H NMR can be recorded in comparison with that of 0 min. Referred to the previous works,49,51 peaks at 3.8 and 4.0 ppm were attributed to H-2,4 and H-3,5,6 of starch. As shown in 2O than that of amylose. Hence, the stronger intensity of α-1,6 bonds was observed than that of α-end group and α-1,4 bond. No other 1H NMR peak occurred until the end of the experiments (seen in Furthermore, 1H NMR spectra of variable heating time at 3 min and 82.5 min as displayed in 52 At the beginning, α-1,6 bonds were stabilized by structural water due to the low degree of gelatinization. This resulted in the strong dipolar interactions, which caused no contribution to the spectrum. However, with increasing heating time, this type of stabilized effect was broken at the high enough extent of gelatinization, which diminished the dipolar interaction. Therefore, the peak of α-1,6 bonds could be detected and intensity was increased with gelatinization time.Interestingly, we compared the It was also found that different processing methods, including varying temperature or extending heating time, showed the significant impact on the details of starch gelatinization. As displayed in 13C NMR spectra at 313 K, 331 K and 340 K for potato and deuterium sample, as well as the sample of potato and NaCl water at 313 K, 328 K and 340 K were measured by liquid state NMR respectively. Due to poor solubility of starch, no NMR signal was recorded in the spectra of both potato starch and potato starch/NaCl sample at 313 K. Importantly, when the temperature was increased to 328 and 331 K, samples of starch and starch/NaCl began to dissolve in D2O respectively. As a result, the peaks of starch could be detected as shown in 13C NMR spectra can be obtained when the temperature increased to 340 K. The 13C NMR chemical shifts for starch were assigned according to the literature.54 Chemical shifts of starch was consistent with that of starch/NaCl during gelatinization, this indicated that no structural changes of starchy backbone occurred with and without adding NaCl. Furthermore, from 55 This result was almost consistent with that of 1H NMR spectroscopy. It indicated that a large amount of starch begins to melt and dissolve in water. As a result, a well resolved 1H and 13C NMR spectra could be recorded under this condition.Furthermore, to better understand the structural changes of starch backbone, 4.1H NMR spectroscopy to in situ monitor the gelatinization process of potato starch at molecular level. VT 1H NMR measurement, from 316 K to 340 K, was measured on the suspension of potato starch and deuterium water with the weight ratio of 1 : 7.5. The hydration level of starch was determined with the increase of temperature. To probe the effect of NaCl on starch gelatinization, VT 1H NMR measurement was carried out on the suspension of potato starch, NaCl and deuterium water. In the presence of NaCl, the initial gelatinization temperature of potato starch was decreased from 331 to 328 K. Meanwhile, in situ1H NMR spectra as a function of time were also carried out to monitor the detailed process during gelatinization. We found that the peak intensity of α-1,6 glucosidic bonds increased obviously with the extending of heating time gradually. Furthermore, the effect with different processing methods during gelatinization, including varying temperature or extending heating time, was investigated in details. Under different treatment, it was verified protons from different group of starch showed the different accessibility for water during gelatinization. In comparison with temperature, gelatinization time as the major factor for acquisition of completely gelatinization was confirmed. All these results indicated that 1H NMR spectroscopy indeed works as a premier tool for in situ monitoring the gelatinization of starch. We expect that this research, as a continuing effort to apply NMR spectroscopy16,56,57 for characterizing starch will pave a new way in the structural elucidation of starch.In conclusion, we have successfully applied The authors declare no competing financial interest.


For 5 and 10 wt.%, no significant differences were observed in mechanical properties at 0° and 90° (p < 0.05). The Vickers hardness values at 0° and 90° from low to high were in the order of 0, 3, 5, and 10 wt.% (p < 0.05). Within the limits of this study, the optimal zirconia filler content in the photopolymer resin for 3D printing was 0 wt.% at 0° and 3 wt.% at 90°.Photopolymer resins are widely used in the production of dental prostheses, but their mechanical properties require improvement. We evaluated the effects of different zirconia filler contents and printing directions on the mechanical properties of photopolymer resin. Three-dimensional (3D) printing was used to fabricate specimens using composite photopolymers with 0 (control), 3, 5, and 10 wt.% zirconia filler. Two printing directions for fabricating rectangular specimens (25 mm × 2 mm × 2 mm) and disk-shaped specimens (φ10 mm × 2 mm) were used, 0° and 90°. Three-point bending tests were performed to determine the flexural strengths and moduli of the specimens. The Vickers hardness test was performed to determine the hardness of the specimens. Tukey’s multiple comparison tests were performed on the average values of the flexural strengths, elastic moduli, and Vickers hardness after one-way ANOVA (α = 0.05). The flexural strengths and elastic moduli at 0° from high to low were in the order of 0, 3, 10, and 5 wt.%, and those at 90° were in the order of 3, 0, 10, and 5 wt.% ( In recent years, computer-aided design and manufacturing (CAD/CAM) technologies have been used in some medical and dental treatments, and the digitization of prosthesis production has been progressing ,2,3,4. PPolymethyl methacrylate (PMMA) has been used as a raw material in several photopolymer resins for 3D printing. PMMA is the most commonly used material in dental prostheses owing to its excellent dimensional stability in the oral environment, low odor, low irritation, low cost, light weight, favorable esthetics, and ease of fabrication and repair. However, several challenges, such as fracture due to water absorption, decrease in flexural and impact strengths, porosity, and polymerization shrinkage, limit the use of PMMA ,6,7. The1/2). Zidan et al. [One study  proposedn et al.  studied 2 nanofiller to a PMMA-based photopolymer resin, and it improved the mechanical properties, thermal conductivity, and thermal stability of the resin. Mohan et al. [2 nanofillers. They evaluated the mechanical properties, filler distribution, and antibacterial activity, among other characteristics of the prepared resins; 1 wt.% of TiO2 and 1 wt.% of PEEK effectively enhanced the mechanical and antibacterial properties of the PMMA composite resin. However, the addition of zirconia [Typically, for a specific range of filler content, a higher filler content implies superior mechanical properties and wear resistance of the composite resin. Therefore, several studies have investigated the addition of micro- and nanosized fillers to photopolymer resins for 3D printing to improve their mechanical properties and overcome their limitations ,13 such n et al.  reportedn et al.  preparedzirconia , a bioceFurthermore, a few studies ,19,20,21This study aimed to evaluate the effects of different zirconia filler contents and printing directions (0° and 90°) on the mechanical properties for 3D printing of the photopolymer resin. The null hypothesis is that there is no difference in mechanical properties due to the difference in the amount of zirconia filler added in different printing directions (0° and 90°).The average particle size was 206 µm, and the highest particle size distribution was between 100 nm and 150 nm . Microscp < 0.05). When the printing direction was 0°, the composite resins with zirconia contents of 0 (control), 3, 10, and 5 wt.% (p < 0.05) were arranged in the decreasing order of their flexural strengths and moduli. The highest flexural strength of 50.6 ± 0.8 MPa and flexural modulus of 0.8 ± 0.03 GPa, which met the ISO standard requirements, were observed for the resin with 0 wt.% filler content. When the printing direction was 90°, the flexural strengths and moduli of the composite resins were in the order 3, 0 (control), 10, and 5 wt.% . The highest flexural strength of 51.0 ± 1.0 MPa and flexural modulus of 0.9 ± 0.04 GPa, which met the ISO standard requirements were observed for the resin with 3 wt.% zirconia content. No significant difference (p > 0.05) was observed between the bending strengths and elastic moduli of specimens with 5 and 10 wt.% zirconia content in both printing directions, and most specimens were fractured into two pieces at the initial stage of the three-point bending test.p < 0.05). When the printing direction was 0°, the composite resins with zirconia contents of 0 (control), 3, 5, and 10 wt.% (p < 0.05) were arranged in the increasing order of Vickers hardness. The highest Vickers hardness of 21.0 ± 0.4 VHN was observed for the resin with 10 wt.% filler content. When the printing direction was 90°, the Vickers hardness values of the composite resins were in the order of 0 (control), 3, 5, and 10 wt.% . The highest Vickers hardness of 21.2 ± 0.4 VHN was observed for the resin with 10 wt.% filler content. No significant difference (p > 0.05) was observed between the Vickers hardness values of specimens with 5 and 10 wt.% zirconia content in both printing directions.The SEM micrographs in In this study, different amounts of zirconia filler were added  to the resin for printing in two different directions (0° and 90°), and their effects on the mechanical properties of the photopolymer resin for 3D printing were evaluated; the flexural strengths and moduli significantly differed under each condition. Therefore, the null hypothesis was rejected. The flexural strengths and moduli of the specimens with 5 and 10 wt.% zirconia filler were significantly lower than those of the control group. When the printing direction was set to 0°, the flexural strength and modulus of the resin containing 0 wt.% zirconia filler were significantly higher than those of the other groups, and when the printing direction was set to 90°, the flexural strength and modulus of the resin composite containing 3 wt.% zirconia filler were significantly higher than those of the other groups. The surface hardness of the specimens may be directly related to the zirconia filler content. These conditions may be used to enhance the mechanical properties of photopolymer resins used for 3D printing, thus emphasizing the significance of this study.The flexural strengths and moduli at 5 and 10 wt.% zirconia filler in the 0° and 90° printing directions were significantly lower than those in the control group by approximately 16% and 10%, respectively. At significantly high filler concentrations, the interaction between the filler particles increases, which may decrease the fracture toughness . During The results of this study were consistent with those of previous studies  reportinSeveral studies ,20,22,27A general-purpose 3D printer was selected for this study, and experiments were conducted using a PMMA photopolymer resin. Several studies have evaluated the physical properties of this resin; herein, a three-point bending test that provides clear interpretation of the results was conducted. Post hoc power analysis was performed using an analysis software . In this study, the significance level was α = 0.05, and the power 1-β was ≥0.8; therefore, the sample size in this study was appropriate. Furthermore, as zirconia, a bioceramic material, has long been widely used in various dental applications, zirconia filler can be added to photopolymer resins for 3D printing, and this may be clinically acceptable in future.The limitation of this study is the use of an unsintered zirconia filler. The properties of a filler-reinforced resin depend on the size, shape, type, and concentration of the added particles . Therefo2O3-stabilized zirconia  was used as the filler to produce composite resins.2O3-stabilized zirconia filler, a preliminary survey was conducted using a photopolymer resin mixed with 0, 1.5, 3, 5, 7, and 10 wt.% zirconia filler. Based on these results and those in the related literature [To determine the optimal mass percentage of the Yterature , the masThe zirconia filler and the photopolymer resin were weighed using an electronic balance . The filler was added to the photopolymer resin. The mixture was stirred for 10 min using a stainless-steel spatula, and the filler was uniformly dispersed in the photopolymer resin. Subsequently, the mixture was poured into the tray of a desktop SLA 3D printer .A 3D CAD software package  was used to design rectangular specimens  and disk-shaped specimens (diameter = 10 mm and thickness = 2 mm), and the output was obtained in the standard tessellation language (STL) file format. The STL data of the specimens were imported to the 3D printing software . The specimens were classified according to the printing directions ; at 0°, 3) according to the ISO standard [A methacrylate-based photopolymer resin  was printed using the 3D printer. After printing, the unpolymerized resin on the surface of the specimens was washed using 99.9% isopropyl alcohol (IPA) for 15 min . After complete drying, the specimens were post-polymerized at 40 °C for 40 min in a post-curing device  under light with a wavelength range of 350–500 nm. After curing, all specimens were wet-polished using a #1200 water-resistant abrasive paper and buffed  using an alumina-based abrasive (particle size = 0.3 µm) for final finishing. The dimensions of the specimens were adjusted for three-point bending tests , and it was calculated using Equation (1):F, l, b, and h denote the maximum applied load (N), support span distance (20 mm), specimen width (mm), and specimen height (mm), respectively, prior to testing.The three-point bending test was performed according to the ISO10477 standard . For theFM, GPa) of the specimens were calculated using Equation (2):F1 and d denote the point load (N) applied on the straight-line portion of the flexural load–deflection curve and the deflection (mm), respectively, at load F1.The flexural moduli (Disk-shaped specimens measuring 10 mm in diameter × 2 mm in thickness were used for the Vickers hardness test (n = 6). A Vickers hardness tester  was used to measure the hardness under a load of 300 gf and a dwell time of 15 s . The finThe particle size and distribution of the zirconia filler and the fracture surface of the specimens after the three-point bending tests were analyzed by scanning electron microscopy . The specimens were attached to an aluminum stub and their surfaces were coated with a thin layer of platinum . The samples were then analyzed at an acceleration voltage of 5 k, a discharge current of 8 μA, and a working distance of 6–10 mm. Microscopic images were acquired at a magnification of 40,000× (10 wt.%) for particle size, 3000× (0 (control), 3, 5, and 10 wt.%) for filler distribution, and 150× (5 and 10 wt.%) for the fracture surface of the specimens after the three-point bending tests. The average particle size was determined based on the SEM images using Fiji-ImageJ software  with the linear intercept method. Particle size distribution was measured for a minimum of 600 particles.p < 0.05) using statistical analysis software .Tukey’s multiple comparison tests were carried out after one-way ANOVA for the average values of the flexural strength, flexural modulus, and Vickers hardness obtained  and printing directions (0° and 90°) affected the mechanical properties of the 3D-printed photopolymer resins.When the printing direction was set to 0°, the optimum mechanical properties that met the ISO standard requirements were obtained at a zirconia content of 0 wt.% (control).When the printing direction was set to 90°, the optimum mechanical properties that met the ISO standard requirements were obtained at a zirconia content of 3 wt.%.Regardless of the printing direction (0° and 90°), adding 5 and 10 wt.% zirconia degraded the mechanical properties of the resins.When the printing directions were set to 0° and 90°, the Vickers hardness increased in the order of 0 (control), 3, 5, and 10 wt.%.Within the scope of this study, the following conclusions can be drawn:


Çatalhöyük is a renowned archaeological site in central Anatolia, best known for its Neolithic occupation dated from 7100 to 6000 cal BC. The site received worldwide attention early on for its large size, well-preserved mudbrick architecture, and elaborate wall paintings. Excavations at the site over almost three decades have unearthed rich archaeobotanical remains and a diverse ground stone assemblage produced by what once was a vibrant farming community. The study presented here adds to our understanding of crops and plant processing at Çatalhöyük by integrating phytoliths and starch analyses on grinding implements found at three domestic contexts attributed to the Middle  and Late  period of occupation. Our results reveal a rich microbotanical assemblage that testifies the use of a wide range of geophytes and wild seasonal resources previously unknown at the site. Moreover, by comparing results from the microbotanical proxies and microscopic wear patterns on artefacts, we are also able to discern various plant processing practices the analysed artefacts were employed for. In sum, this work further expands our understanding of plants and crop processing activities performed by the inhabitants of Neolithic Çatalhöyük. Archaeobotanical remains are the best proxies for studying the emergence of prehistoric agricultural practices. The informative potential of such remains, coupled with ethnographic and ecological models, allowed researchers to identify diverse plant processing strategies of past societies –5. WithiExcept for some exceptional finds e.g., , 17), it, it17]),Significant advances in the functional analysis of grinding tools during the last two decades have revealed a wealth of information about the function of tools and subsistence practices more broadly , 31–35. Phytoliths originate from the deposition of opal silica in plant cells and cell walls creating casts of the cells or entire tissues. Grasses are the most prolific producers but phytoliths are found in many other plant groups with species of economic interest . Since pPhragmites spp.) and sedges (Cyperus spp.), combined with suitable dry areas for farming )nomina conservada from ICPN 1.0), and identification was achieved using published material  were recorded separately. Concentrations of phytoliths per gram of acid-insoluble fraction (AIF) were calculated following Albert et al. , and morl e.g., , 129) an annominaFollowing the technological analysis of the ground stone assemblage a number of objects were selected for microwear analysis, including the set of tools that were further sampled for microbotanical analysis. The combination of microwear and microbotanical analyses discussed in this paper allowed not only for a better understanding of the plant exploitation strategies but also for a more nuanced appreciation of tool function at the site , 90. The® Light Body Coltene and Provil® Novo Light Regular Set Heraeus Kulzer), which can accurately replicate the micro-topography of the tool surface, were taken. Recorded wear patterns included grain edge rounding, levelling, grain extraction, micropolish features, texture and development, microstriations, microfractures, and the presence of residues . We have made a short mention in the test of these plants, their medicinal uses and how these are processed.Section 4.1, lines 575- 579.It is worth noting that some Liliaceae and Iridaceae species contain toxins and need to be processed for detoxification before consumption, or they are used in small quantities for medicinal purposes. This is the case for species of the genus such as Fritillaria, Lilium and Tulipa, whose USOs can be crushed and powdered and used in folk medicine in parts of current Turkey for treating wounds and other ailments .Regarding how these could have been processed in the past, many of the underground storage organs mentioned in the paper can be consumed or processed fresh. But, considering the hardness of some of these, a pounding, rather than grinding process could be assumed. However, this is not always the case, underground storage organs can be dried first, and then ground, such as the case of orchid bulbs used for making Salep, a starchy flour commonly consumed in Turkey in beverages and desserts. Other practices include boiling, roasting, or even fermenting that make the tubers softer thus removing the need for pounding. These practices would affect the way such plants were processed and the kind of reduction technique and tools used. We ignore whether these practices were also performed at the site, and in the case of underground storage organs, and despite the obvious assumptions that can be made based on the tools' typology, the starch data can only tell us that these plants were subject to a reduction technique which involved the tool where the starch was found. Besides, there is also an etymological consideration involved in the way we interpret the tools uses, and the reduction technique that was used at the time of processing the USO. This is related to the intrinsic definition of a grinding stone itself, that links an object type with a very specific practice. We are aware that this is not always the case, these are dynamic objects, involved in multiple functions and reduction techniques that expand the plant-related activities as it has been demonstrated in its section. In sum, we recognize the great interpretative potential that can emerge by going into detail about the plant's characteristics that influence the ways it could be treated, and the reduction technique used. But we are also aware that the information provided by the tools is a palimpsest of multiple plants, uses, and activities, and based on the arguments presented above we prefer to be careful and to not overextend the interpretative potential of our finds. • On another note just a couple of edits:• Table 3 ‘smoot’ should presumably read ‘smooth’• Table 4 need to specify what R representsThese mistakes and editions have been addressed. Reviewer #2: This concise and very interesting article tackles the question of the function of the ground stone implements uncovered at the Neolithic site of Çatalhöyük. Although the collection has been previously presented in various papers, the question of tool function was mainly indirectly addressed so far, through morpho-typological analysis, using ethnographic analogies and/or spatial distribution. Thus, previous interpretations often posited grinding and pounding implements as a good proxy for plant processing and/or placed an emphasis on the multi-functional character of the tools. I believe that one of the great interests of the article is to go beyond such generalizations and to provide a direct, multi-approach investigation of ground stone tool function. The article presents in detail phytoliths and starch types extracted from the surface of a sample of tools. This is later compared with the results of a use-wear analysis, which are more briefly discussed. In the conclusion, the authors emphasize that their results provide additional evidence for a great diversity of plants being exploited at the site. The contribution of wild plants is also underlined, among which geophyte species and panicoid grasses which were seldom documented so far. Although these conclusions are well supported by the data presented, it is nevertheless based the analysis of a very limited sample of tools. In general, I believe that the authors should provide more information on their research and clarify various points so the significance and limits of the results presented can be more clearly assessed by the readers. Suggestions are provided bellow:• One of the main points not addressed here is sample size. Use-wear and residue analyses are very time-consuming, still the fact that the study focuses on only 8 out of more than 2000 artifacts (yet less if we count only on grinding and pounding implements) should be more clearly acknowledged, discussed and taken into account in the discussion;We recognize the concern about the sample size. However, the artifacts were selected because when we started this study, there were among the most complete artifacts since complete tools are not that common on the site . In addition, most of the tools were extensively handled, minimizing the sample size due to the risk of some artifacts have being exposed to modern starch contamination. We also realized that these artifacts were found on very important and well-studied buildings and contexts providing an opportunity to integrate our results into further and multidisciplinary discussions about the activities performed on buildings 80, 52 and 44. Nonetheless, despite being a small sample, we believe that by implementing a multi-proxy approach, we produced a more robust and complex set of data than the one we would have obtained by analyzing more artifacts with only one proxy.To conclude, we believe that the small sample size does not hamper the informative potential of these artifacts as has been highlighted in the paper. The main point of the paper was not to generalize on how grinding tools were used at the site but to demonstrate the research potential of combining two microbotanical proxies with microwear analysis. We believe that this was successfully achieved based on the diversity of plants identified, including wild plants processed on-site that had not been previously identified in the macrobotanical record. In this sense, this is not an attempt to deduce the scale of processing and reliance on these plant resources nor variations between periods or different buildings, which would require a more robust sample. But we recognize the need of acknowledging the sample size as suggested and has been addressed in the paper.Section 5.0, lines 770 – 772.Despite that the tools examined represent only a small portion of the whole grinding tool assemblage at the site, our work made evident the wide spectrum of plant resources, both domesticated and wild, that the people of Çatalhöyük were using.Further discussion below.• Along the same line, the sampling strategy should be presented in more detail. From the description of the tools, it does not seem that the best-preserved artifacts were selected which is puzzling, especially because the methods employed encompass use-wear analysis.Some of the reason that influenced the sampling criteria has been presented above. But we have made changes addressed this on the paper.Section 2.4, lines 277 - 287Following the technological analysis of the ground stone assemblage a number of objects were selected for microwear analysis, including the set of tools that were further sampled for microbotanical analysis. The combination of microwear and microbotanical analyses discussed in this paper allowed not only for a better understanding of the plant exploitation strategies but also for a more nuanced appreciation of tool function at the site . The selection of material was guided by typological, technological and contextual criteria, as well as the overall condition of the material. While tools that survive in generally good condition without visible alterations were favoured, in some cases tools that show low or moderate heat alterations but derive from significant contexts  and/or survive intact were also included in the sampled material. In cases where the tools exhibited more intense burning, which affected the use-faces of the implements, these were not subjected to microwear analysis. • While the procedure, reference collection, limits in interpretation are well presented for phytoliths and starch, less details are provided for use-wear approach. More information about the reference collection used, some photographs of the experimental tools would be appreciated.We created a separate section on Microwear analysis in Materials and Methods , where we included more information about microwear analysis, the experimental reference collection, and created a new figure with experimental wear traces.• Concerning the results of phytoliths and starch analysis: Were sediment samples only examined for starch? Each sediment sample was subjected to both, starch and phytolith analyses as has been presented in the section 2.2 Sampling and laboratory procedures for microbotanical analysis.Section 2.2, lines 250 – 253.The extraction procedure followed a combined protocol for phytoliths and starch extraction adapted from Pagan-Jimenez [59] and García-Granero et al. [112]. In this method, starch grains are first extracted from the sediments to avoid their exposure to any stronger chemical used for the phytolith extraction [113–115].We also modified the Taphonomy and contamination section accordingly.Section 3.1.2, lines 469 – 470.Control sediment samples were rich in phytoliths and devoid of starch except for Building 44 where the sediment produced 9 heavily damaged starch grains.• Is this because tool contamination was not found to be an issue for phytoliths? Despite this being a problem that applies to both proxies, there is always more concern about the starch so much so that a few researchers have doubts over the preservation of starch in archaeological sediments/artefacts. Therefore, when working with ancient starch there is always concerns whether the starch recovered is ancient or a result of modern contamination. This topic has been widely discussed, but the assumption is that ancient starch survived trapped in the tool's porous surface. This, assumption is supported by the fact that the control sediments do not present starch granules or only a few. Besides the check provided by control sample, we proposed a novel solution by correlating the most predominant starch damage with the tool's depositional or post-depositional context. To be more precise, those that were exposed to fire vs those that were not.• It is intriguing and noteworthy that sediment samples were very poor in starch while in situ plant processing is suggested for 2 of the buildings sampled here, I believe this should be discussed. In-situ plant processing was only suggested for the quern embedded in the floors of building 80 because this was a fixed installation. We discussed this artifact and the interpreted in-situ plant processing in section 4.3.1. Artifacts from building 44 were deposited in a cluster and those from building 52 were in a storage room, thus we cannot nor are suggesting that these were used in-situ and their mobile nature has been discussed on the paper as well in Section 4.3.2. We believe the confusion could be caused by sentence on the original manuscript;This in situ assemblage related to plant storage suggests a diversified diet [21], and the spectrum of plant use in this building was probably even wider than what the macrobotanical evidence suggests [80].We have modified this sentence;Section 2.1, line 185 - 188This assemblage related to plant storage suggests a diversified diet [21], and the spectrum of plant use in this building was probably even wider than what the macrobotanical evidence suggests [80].This was also addressed and clarified in Section 4.3.2, lines 666 – 669.The very low presence of starch grains on the tool (n=11) and the complete absence of starch from the surrounding sediments (see [246]), together with the phytolith evidence, strongly support that this implement was used in situ for dehusking or beating spikelets for separating seeds from the chaff in cereals .• It is my understanding that damages on starch are mainly attributed here to post-depositional processes, but this should be more clearly stated. Concerning the damage on starch as being caused by post-depositional process (fire), it is mentioned in the paper, and its interpretation is only limited to strengthen its origin as ancient starch. Section 3.1.2, line 489 – 490.These observations are useful as they confirm that the recovered starches are of secure provenance and not the product of modern contamination.Further discussion below.• The question of gelatinization and the fact that it may occur during post-depositional burning is very interesting. Are the authors referring here to the same type of starch grain damages than the ones ascribed in a few papers to beer production, or are those rather described as ‘full gelatinization’ which they do not found in Çatalhöyük ground stone tool sample? It would be useful to make this point clearer. Starch damage can indeed be very useful to identify certain practices such as fermenting, boiling, etc. It's possible that some of the observed damage was caused by the type of processing to which the plants were subject, such as the case for broken or cracked starch grains, traces that occur while grinding and pounding. I believe here the confusion could be cause by what is causing the starch gelatinization either partially or full. Heat treatment is the principal anthropic cause of gelatinization on starch, in our case, this could be caused by processing practices such as roasting or parching the grains before grinding. However, enzymatic activity can also cause starch gelatinization, produced when the grains are left to sprout  before being processed or by microbial activity (fermenting). However, making such interpretation is very complex, not all starches are equally resistant to heat, enzymes, or mechanical damage and we do not have enough information to support any of the mentioned treatments, especially since some artifacts were exposed to fire. In addition, apart from differentiating from the heat environment (wet or dry), starch gelatinization can not be exclusively linked to a specific treatment [2–4]. Instead, its interpretation relies vastly on the archaeological context .Concerning our interpretation of the exposure to fire as the cause of starch gelatinization, we consider that this exposure restrict any attempt to link any observed starch damage to a particular processing activity. Therefore, we can only hypothesize that starch grains that were already trapped in the tools started to partially gelatinize when the artifacts were exposed to heat once the buildings 80 and 52 burned down. Following the reviewer suggestion, we have made this point clearer. Section 3.2, lines 486 – 489.Therefore, gelatinization-like damage in our samples in all probability is not related to intentional food processing activities or cooking, but to post-depositional processes such as the artefact’s exposure to heat when both buildings caught fire. • Phytoliths modifications are not discussed while this could also have contributed to explore plant processing, the authors should specify this and maybe explain why.This is a very important observation. Despite that it is not as informative or evident as in the case of starch, certain mechanical processes like threshing or dehusking could cause straight cuts or breaks on silicified conjoined cells. These could be used as a proxy for post-harvesting processes. We have made changes to the text and included this.Section 3.2, lines 526 – 527.In some cases, silica skeletons presented straight transversal cuts , probably caused by post-harvesting processes such as trampling and dehusking .• Regarding the results of the use-wear analysis: Given the state of alteration mentioned for some of the tools, an assessment on use-wear preservation should be provided. Clarification about heat alterations was provided in Section 2.1.and in section 2.4. As noted in Table 1 the two tools with moderately/heavy degree of burning were not subjected to microwear analysis. • In several research, criteria have been suggested to differentiate various types of plants or processing for instance greasy/versus non greasy plants, cereals/legumes, dehusking/grinding , yet here the authors are identifying ‘plant processing’. Why does the analysis remain at such a general level and do not further assess the properties of the matter processed? It may be due to the compressed versions of the images, I have but it is difficult to see any micropolish in Fig10 e and f. In addition, given the variability observed in use-wear associated with the processing of various types of plant, the concept of ‘plant micropolish’ is problematic (as there are different types of micropolish associated with plant processing). Furthermore, stone on stone contact has been reported to occur during grinding (see for instance Dubreuil 2004) so its development can be concomitant to other type of wear.We included a paragraph in Section 2.4, lines 309 - 330 that discusses the difficulty in differentiating types of plants in archaeological materials that display mixed wear signatures and are associated with the processing of different contact materials and have included in section 4.0 Discussion more information about the type of plant processing when this level of interpretation was possible. In the case of tools with evidence for use against multiple materials and mixed wear signatures it is not possible to interpret the type of plant contact material in more detail. We agree about the stone on stone contact which we also have identified during the microwear analysis of some of the grinding tools (see Table 1) and we clarified this point further in section 4.3.2, lines 707 – 709.• Fig 10 B (and in the text as well) mention the processing of ‘ woody’ material, does the authors mean abrasion or grinding? It seems that the use-wear would fit with those associated with a use as an abrader but if the authors mean grinding, could they provide more explanation about the type of woody matter and for what purpose? We addressed this point in Section 4.3.2 and lines 736-744. In the case of the woody plant materials, based on the characteristics of the use-face of the tools (intense levelling and smoothing that extends across the whole surface of the tool), their presence on the tool surfaces is more consistent with grinding activities. The presence of wood and wood-like materials has also been noted in other studies of grinding tools . Hamon and colleagues [137] have suggested that the grinding of wood and bark materials may be associated with the acquisition of powdered materials with colouring or medicinal properties, or potentially fruit processing. Continuing microwear analysis of the Çatalhöyük grinding tools may shed more light on the type of processing associated with this type of contact material.• Finally, it seems that part of the tool life-history is described in the section presenting the artifacts, with no or minimal supporting information regarding how those conclusions were reached, while in fact this life-history reconstruction should be part of the use-wear analysis.The detailed technological, spatial and contextual analysis of the Çatalhöyük ground stone assemblage has been presented in other publications and we included in the text in section 2.1. the following passage to reflect this: Line 134 - 137: Detailed technological, typo-morphological and contextual study of the Çatalhöyük ground stone assemblage, the results of which are presented in detail elsewhere , provides important insight in the life histories of these artefacts and the social practices that surround their use and discard by this Neolithic community.In addition, we have provided some additional information about the tools in the text in section 2.1. which should be read in tandem with the information given in Table 1.• Overall, the article is well illustrated. One table with synthetic presentation of the residues and use-wear results for each stone is missing, so both approaches could really be integrated. Providing more illustration of use-wear, of the experimental collection and surfaces of the tool at low magnifications would also strengthen the interpretation suggested. Finally, I would suggest including a few figures combining the various faces of the tool, use-wear and residues found on the same object.We appreciate the suggestion and have included a collage picture showing the artifact along with use-wear traces and micro-botanical remains on Section 4.3.2, line 714.Reviewer #3: This is a very clearly written and exciting paper that reports on the first integrated phytolith, starch and use-wear results from ground stone implements at Catalhoyuk. The results are explained with admirable clarity and with full reference to the relevant literature. • I have only one, relatively minor, comment. Regarding the Panicoid grasses and C4 isotope discussion, the macrobotanical assemblage does contain some other C4 grasses  that were also observed in intact sheep/goat pellets, and so likely contribute to both the caprine C4 signal and the human C4 signal. This does not exclude that other, Panicoid grasses also contribute to the human C4 signal, but it should be made clear in the text that there are other C4 taxa that likely contributed in any case.We agree on this observation and changes has been made on the text on Section 4.2, lines 635 - 646).Bibliography1. Bozyel ME, Merdamert-Bozyel E, Benek A, Canlı K, Altuner EM. Ethnomedicinal Uses of Colchicaceae and Liliaceae Taxa in Turkey. In: Christov I, Krystev V, Efe R, editors. Advances in Scientific Research: Engineering and Architecture. Sofia: St. Kliment Ohridski University Press; 2020. pp. 649–660. 2. Pagán-Jiménez J. Evaluando algunos mecanismos de conservación/degradación en almidones modernos por medio de ensayos y experimentos controlados que replican ciertas formas antiguas de procesamiento y cocción de órganos almidonosos. Elaboración de dos tipos de chicha de maíz: chicha fermentada con saliva y otra con levadura. Quito: Instituto Nacional de Patrimonio Cultural; 2015. doi:10.13140/RG.2.1.5089.74403. Pagán‐Jiménez JR, Guachamín-Tello AM, Romero-Bastidas ME, Vásquez-Ponce P. Cocción experimental de tortillas de casabe (Manihot esculenta Crantz) y de camote (Ipomoea batatas [L.] Lam.) en planchas de barro: evaluando sus efectos en la morfometría de los almidones desde una perspectiva paleoetnobotánica. Americae. 2017;2: 27–44. 4. Chantran A, Cagnato C. Boiled, fried, or roasted? Determining culinary practices in Medieval France through multidisciplinary experimental approaches. J Archaeol Sci Rep. 2021;35: 102715. doi:10.1016/j.jasrep.2020.1027155. García-Granero JJ. Starch taphonomy, equifinality and the importance of context: Some notes on the identification of food processing through starch grain analysis. J Archaeol Sci. 2020;124: 105267. doi:10.1016/j.jas.2020.105267AttachmentResponse.docxSubmitted filename: Click here for additional data file. 14 May 2021A microbotanical and microwear perspective to plant processing activities and foodways at Neolithic Çatalhöyük.PONE-D-21-00753R1Dear Carlos Gabriel Santiago-Marrero,We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements.Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication.http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org.An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at onepress@plos.org.If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact Kind regards,Eleni Asouti, Ph.D.Academic EditorPLOS ONEAdditional Editor Comments :Please ensure that your tables are adequately formatted for inclusion in the manuscript as it will appear on the journal website and in pdf form. I still have some concerns regarding the size and formatting of Table 1, including its individual cells (parts of the text are missing and/or are illegible in the .pdf of Table 1 you submitted as a separate attachment). Please read very carefully all the instructions included in the author guidelines and liaise closely with the PLOS ONE editorial team. It will not be possible to fix any formatting issues resulting in parts of Tables being cropped or illegible after publication. Please also note that it is your responsibility to proof read thoroughly all text, tables and captions to correct any residual typos, when you receive your proofs from the PLOS ONE journal office. 2 Jun 2021PONE-D-21-00753R1 A microbotanical and microwear perspective to plant processing activities and foodways at Neolithic Çatalhöyük. Dear Dr. Santiago-Marrero:I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. onepress@plos.org.If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact plosone@plos.org. 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This systematic review aims to evaluate the different pretreatments of the zirconia surface and resin cement in order to determine a valid operative protocol for adhesive cementation. Methodologies conducted for this study followed the Prisma  guidelines. An electronic search was performed in four databases. The established focus question was: “What type of surface conditioning method is the one that obtains the best adhesion values to zirconia over time by applying a resin cement?” Forty-five relevant papers were found to qualify for final inclusion. In total, 260 different surface pretreatment methods, mainly combinations of air-abrasion protocols and adhesive promoters, were investigated. Altogether, the use of two artificial aging methods, three types of cement and four testing methods was reported. The results showed that mechanicochemical surface pretreatments offered the best adhesive results. Self-adhesive cement and those containing 10-MDP obtained the best results in adhesion to zirconia. Artificial aging reduced adhesion, so storage in water for 30 days or thermocycling for 5000 cycles is recommended. A standardized adhesive protocol has not been established due to a lack of evidence In recent decades, the increasing aesthetic needs in dentistry have led to the progressive overcoming of metal-ceramic prostheses and led to a focus on indirect metal-free restorations. Yttrium-stabilized zirconia has occupied an increasingly important role and offers a wide variety of clinical applications, such as root posts, implant abutments or as a material of choice for indirect ceramic restorations. It has the most favorable mechanical properties compared to other high-strength ceramics with flexural strengths of 700–1200 MPa, fracture resistance of more than 2000 N and fracture toughness of 7–10 MPa ,4. HowevSurface treatment with hydrofluoric acid (HF) and silane coupling agent application of the silica-based ceramics is a well-established method to achieve durable adhesion to resin-based materials . HoweverSeveral methods have been used to evaluate the bond strength of resin-based materials to dental ceramics, including macroshear, microshear, macrotensile and microtensile tests. Furthermore, methods to evaluate bond durability simulating the oral conditions include short- and long-term water storage and thermocycling at diverse temperatures, dwell time, and number of cycles. Therefore, it is difficult to compare different studies on the same materials even when the same test method was employed ,30.Due to the great increase in in vitro studies in recent years and the lack of consensus on resin-bonding protocols for zirconia restorations, it is necessary to evaluate the current data to unify criteria and provide clinicians with relevant information for their daily activity. Therefore, the aim of this study was to systematically review the literature to evaluate the different zirconia surface pretreatments and resin cement to determine a valid clinical protocol for adhesive cementation.The present systematic review followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The praThe literature search was performed by two independent reviewers , including articles published between 1 April 2015 and 1 December 2020, and any disagreement was resolved by a third reviewer (M.J.S.). The electronic databases screened were PubMed, The Cochrane Library, Scopus, and Web of Science (WOS). The following search terms and their combinations were employed: “zirconium,” “zirconia,” “3Y-TZP,” “3Y-TZP ceramic,” “dental adhesion,” “dental bonding,” “bond strength test,” “cement” and “resin bonding.”The articles included followed the inclusion criteria: published in English, in vitro studies that reported on adhesion to zirconia restorations by resin cement, using microtensile, macrotensile, microshear or macroshear bond tests with results (mean and standard deviation) in MPa and subjected to artificial aging. Studies with the following criteria were excluded: studies on high translucent zirconia, data were not presented in MPa or without normal distribution, studies with a number of specimens less than 5, studies with samples subjected to less than 5000 thermocycles (TC), or less than 1 month of storage. Clinical trials, case reports, case series and pilot studies were also excluded. Studies that did not specify any of these data were removed from the review. Any disagreement regarding the eligibility of the included studies was resolved through discussion and consensus. The inclusion and exclusion criteria are listed in The following variables were extracted: mean and standard deviation of the bond strength recorded in MPa, type of pre-treatment technique, type of cement, methods of aging (storage conditions and duration and/or the number of thermocycling procedures) and type of bond strength test.The research carried out in PubMed , CochranIn the selected 45 articles, a total of 511 experimental groups were identified with bond strength results (MPa). In these groups, the search identified 260 different methods of zirconia surface pretreatments prior to adhesive cementation. Of the selected 45 articles, 43 articles did not report a control group with no conditioning surface. In the other two articles, two experimental groups were identified as control groups. Pretreatment techniques were listed as described by the authors and classified into three groups: Mechanical conditioning methods using abrasives were found in 140 experimental groups; chemical methods using adhesive promoters were used in 5 experimental groups; mechanicochemical conditioning methods based on using abrasives or etchants followed by adhesion promoters were practiced in 364 experimental groups. The main zirconia pretreatments are summarized in Resin cement is summarized in Regarding the aging conditions, the type and duration of aging methods were recorded. In the present systematic review, the studies were conducted in vitro, and a great heterogeneity was observed among the studies. Despite the limitations of this type of study, it is necessary to evaluate the behavior of different materials and techniques before their clinical application ,32,33.In this systematic review, pretreatment techniques were classified into three groups: (1) mechanical: studies that used air-abrasion protocols, laser, ceramic coating, or chemical etching, (2) chemical: studies that employed coupling agents such as adhesive resins, silanes or primers, (3) mechanicochemical: when both mechanical and chemical conditioning methods were applied. Control groups were defined as zirconia substrates with no surface pretreatment.The studies agree that the zirconia surface needs to be prepared before applying the resin cement since all the pretreatments increased the bond strength, improving the values of the control group ,35,36. TSeveral mechanical pretreatments have been investigated. Sandblasting with alumina particles improved the bond strength values due to the increase in surface energy, wettability, roughness, and the appearance of hydroxyl groups that will facilitate bonding with the primer/universal adhesive/cement ,41,43,482), with different parameters of power, energy intensity, distance, and duration. Most of the studies concluded that the application of laser did not increase the bond strength compared to sandblasting and did not obtain acceptable adhesion values [The application of lasers to the surface of zirconia is based on the same principle as sandblasting, obtaining a rough surface and an increase in wettability that allows micromechanical retention with the resin . Differen values ,40,43,51n values ,43.An electrical discharge machine (EDM) described by Rubeling et al.  was usedRecently, plasma has been introduced as zirconia surface pretreatment to increase the surface energy and optimize the chemical surfaces of the substrates without affecting its structural properties. However, the application of oxygen or argon plasma did not obtain good adhesion values after artificial aging, which added to the appearance of impurities on the surface of the zirconia, indicated its susceptibility to hydrolytic degradation ,55,71.Even though zirconia is a polycrystalline ceramic that cannot be etched due to the absence of a silica phase, different etching methods have been tested in acidic solutions such as phosphoric, nitric and HF acid. These solutions showed contradictory results compared to sandblasting ,61,63,68Several ceramic coating methods have been also investigated. It has been reported that the application of a layer of silica glaze creates a more reactive and etchable glass surface that can be treated as a glass-ceramic. The posterior HF etching removes the glassy matrix creating a porous surface with high surface energy, ideal for cement penetration . This prTo simplify the steps in the adhesion and make life easier for clinicians, the adhesives started to contain chemical promoters, being called universal. The objective was to achieve the preparation of the restoration without the need to add another component. Most of these universal adhesives contain 10-MDP at different concentrations and have been the most studied products in the last five years. It has been reported that the application of a universal adhesive with 10-MDP increased adhesion to zirconia after sandblasting ,50,55,69In the reviewed articles, most of the studies involved mechanicochemical methods ,75,76,77TSC consists of zirconia sandblasting with silica-coated alumina particles. The impact of the particles creates an irregular surface incorporating silica into the zirconia structure and allowing the use of silane as a binding agent to silica and resin. This leads to the appearance of chemical chains of siloxane between cement and residual silica, increasing adhesion and improving the wettability and surface energy of zirconia ,76. The Feldspathic ceramic sandblasting and silane application appears to be a promising surface pretreatment, showing statistically better values than the TSC plus silane after thermocycling . ThammajOther methods to silicatize the zirconia surface have been introduced. The Silano-Pen system consists of a lighter with a solution of butane and silane. When butane is burned, the silane compound decomposes into SiOx-C fragments that adhere to zirconia, allowing it to be silanized. However, its effectiveness has not been proved compared to other methods .The classification of resin cement was complicated because of the great variation in their chemical compositions: phosphoric acid esters, 10-MDP, HEMA, glycerolphosphate dimethacrylate (GPDM), 4-META, bis-GMA or triethylene glycol dimethacrylate (TEGDMA). In addition, the exact composition or percentage of each component is hardly shown due to the lack of information from manufacturers. Therefore, their classification was structured in self-adhesive, cement with 10-MDP, and Bis-GMA cement (without 10-MDP or were not self-adhesive). In general, within the same group, the cement had great variability due to both the percentage of the different components and the viscosity of the cement, which can interfere with micromechanical interpenetration . There iDifferent types of tests have been used to assess the bond strength between zirconia and composite cement that can be explained by the lack of an international standard. The most used was the macroshear test, probably due to its simplicity of use. Otani et al.  describeThe most used method for artificial aging was liquid storage and thermocycling. Liquid storage allows the evaluation of hydrolytic degradation, and thermocycling reproduces in vitro hydrothermal aging ,78. The Since this systematic review was based on in vitro articles, it was not possible to perform clinical guidelines because there are certain clinical factors, such as saliva contamination or parafunctional habits, that negatively affect the adhesion . Further-There are a great variety of zirconia surface pretreatments, cement, artificial aging method and tests used in the studies that make it difficult to compare the results.-Zirconia surface cleaning must be performed before pretreatment methods to adhesion.-Mechanicochemical surface pretreatments offered the best adhesive results. Tribochemical silica coating at a pressure of 1.8–2.8 bar has proved to achieve a significant increase in adhesion to zirconia.-New methods as feldspathic ceramic sandblasting and silane application or YAG laser combined with silane seem to be promising alternatives in adhesion to zirconia.-There is great variability in the percentage of components and the viscosity of the resin cement. Self-adhesive cement and those containing 10-MDP obtained the best results in adhering to zirconia, without clarification of which is the best.-The use of a 10-MDP primer is still controversial.-Standardization of test to evaluate the bond strength between zirconia and resin cement is needed-Artificial aging decreased adhesion; therefore, storage in water for 30 days or thermocycling for 5000 cycles must be performed in laboratory studies.-A clinical protocol for adhesive cementation to zirconia has not yet been performed.Within the limitations of this review, the following conclusions were drawn:


As I write this Editorial at the end of September with the new academic year about to start, I realised that I have done nearly 3 months as Editor in Chief of Engineering in Medicine. It has been an interesting, stimulating and educational time. I would like to bring you, our readers, up to date with the changes.We have renewed the Editorial Board. Three people have ceased to be Associate Editors, Klaus Radermacher and Georges Limbert have decided to step down, but will remain members of the Editorial Board and I will cease to act as an Associate Editor. I would like to thank both Klaus and Georges for their efforts over many years working for Engineering in Medicine, many papers have benefitted from their thoughts and input. I certainly appreciate the time they have put into the journal even if it is not so obvious to the Readers. We have now recruited eight new Associate Editors and you can see them listed in the front cover of the journal and on the journal web page. We previously had only one Associate Editor from China and the Far East and we have recruited Duanduan Chen from Beijing, Eijiro Maeda from Nagoya, Japan and Dalton Tay from Singapore to spread our influence to the East. In the UK we have recruited Damien Lacroix and Koushik Maharatna to broaden our expertises including modelling Artificial Intelligence and Machine Learning. We have recruited Magnús Gíslason from Iceland and Heidi Ploeg and Carolyn Sparrey from Canada going westward again bringing in new expertises. We now have two more women as Associate Editors and I hope that you will consider we have increased our ethnic diversity. I also hope that by reducing the workload on all the Associate Editors we have enabled them to process papers more effectively.On the Board we have had six retirements, John Fisher, Garth Johnson, David Mitton, Robert Reiner, Antonio Strozzi and Mark Taylor. They have been on the Editorial Board for many years each and we will miss their wise council in meetings. We have our first ever members from Africa and the Middle East and I’m delighted to thank Elsie Kaufmann and Ali Al-Timemy for joining us and expanding our geographical spread. They are joined by Laoise McNamara and Nicholas Dunne from Ireland, Thomas Schauer from Germany, Elizabeth Clarke from Australia, Cheryl Quenneville from Canada and Deepak Vashishth from USA. Consequently, along with being the only one of the 16 journals of the Proceedings of the Institution of Mechanical Engineers with a female Editor in Chief, a quarter of the whole Editorial Board are women.However, I am delighted to point out stability and an extraordinary degree of devotion to Engineering in Medicine, Peter Walker remains on the Editorial Board having been recruited at the start of the journal about 50 years ago, thank you Peter, for a half century of support . I should also say that two of the new recruits reported that, like me, their first published paper was in Engineering in Medicine. We hope that the Editorial Board combines a track record with novel ideas and publishing excellent papers.We will be looking to recruit further new Board Members next year to ensure that we have the breadth of expertise and geographical spread so that the next half century is one of growth for Engineering in Medicine, however journal publishing changes.Queen Mary University of London, London, UKLiz Tanner