Polygraphus proximus Blandford (Coleoptera: Scolytinae) is a non-aggressive phloephagous bark beetle that feeds on Far Eastern firs. They have caused mass mortality in Russia and Japan. However, the distribution pattern of entry holes of P. proximus and spatial relationships with bark characteristics have not been studied. Thus, we investigated the distribution pattern of entry holes of P. proximus. The distribution of entry holes was significantly uniform in most cases. As the attack density increased, an aggregated distribution pattern within a short distance (< 4.0 cm) was observed. The rough bark had a significantly higher number of entry holes than the remaining bark. The distribution pattern of entry holes demonstrated a significantly aggregated spatial association with rough bark. Finally, rough bark around knots had significantly thicker phloem than the remaining barks. These suggest that P. proximus may preferentially attack rough bark to reproduce in the thicker phloem under a rough bark surface.Bark beetles attack their hosts at uniform intervals to avoid intraspecific competition in the phloem. Bark texture and phloem thickness also affect bark beetle attacks, and the bark characteristics are not spatially homogeneous; therefore, the distribution patterns of entry holes can demonstrate an aggregated distribution.  Most bark beetles reproduce in the phloem tissue of woody plants. Their adults land on a tree, bore into the phloem, copulate, and excavate galleries along which they oviposit. The larvae feed and develop as they construct galleries in the phloem. In the bark, a higher density results in severe intraspecific competition for nutritious phloem among larvae and low reproductive success –3. AccorBoth outer and inner bark characteristics can also affect bark beetle attacks. For instance, several bark beetles preferentially attack a certain surface  texture, such as rough texture and crevices –9. PhloePolygraphus proximus Blandford (Coleoptera: Scolytinae) feeds on Far Eastern firs. They infest fresh-cut logs and trees weakened by fire, pathogens, typhoons, or defoliation during the endemic phase in their native range [P. proximus is the most destructive pest in natural Abies sibirica Ledeb. forests [A. firma Sieb. et Zucc trees in Japan [A. veitchii Lindl and A. mariesii Masters trees have been observed in Japan [The non-aggressive phloephagous bark beetle ve range –14. It hve range . In Sibe forests ,17. Theyin Japan . Recentlin Japan –21.P. proximus has not been demonstrated. We hypothesized that rough bark had thicker phloem within a species, and the distribution pattern of entry holes of P. proximus was accordingly aggregated on the rough bark. The distribution pattern of entry holes can influence population dynamics via reproductive success in the bark [P. proximus and the spatial association between entry holes and bark surface roughness. The aim of the present study was to determine 1) the distribution pattern of entry holes of P. proximus, 2) the spatial association between entry holes and bark roughness, and 3) the association between bark surface roughness and phloem thickness.To date, the distribution pattern of entry holes of the bark ,23. ThusThe following institutes granted permission of the field surveys and samplings: The University of Tokyo Hokkaido Forest, The University of Tokyo; The University of Tokyo Chiba Forest, The University of Tokyo; Sugadaira Research Station, University of Tsukuba; Yatsugatake Forest Station, University of Tsukuba; Education and Research Center of Alpine Field Science, Shinshu University.Polygraphus proximus feeds on the following Far Eastern fir species: Abies firma, A. holophylla Maxim., A. homolepis Sieb. & Zucc., A. mariesii, A. nephrolepis (Trautv. ex Maxim.) Maxim., A. sachalinensis (Fr. Schmidt) Masters, A. sibirica, and A. veitchii [veitchii ,13,24. Tveitchii ,16. The veitchii ,26. Theiveitchii . The femveitchii ,16,25. EAbies species are coniferous evergreen trees that grow to a height of 25–30 m. Five Abies species are native to Japan. A. sachalinensis is native to the Sakhalin and Kuril Islands, Russia, and Hokkaido Island, Japan. A. veitchii is native to Honshu and Shikoku, Japan. It dominates mountain forests at elevations of 1500–2500 m [A. mariesii is native to the mountains of central and northern Honshu, Japan. A. firma is native to central and southern Japan. A. homolepis is native to the mountains of central and southern Honshu and Shikoku, Japan.0–2500 m . A. mariAbies species were felled in April and May 2019 (Four to eight un-infested trees (diameter of breast height: 14–20 cm) of each of the five May 2019 . The treAbies spp. are surrounded by rough texture  function analysis using R 3.6.1 [r) function is a tool for analyzing spatial point process data [r) is defined as the expected number of entry holes within distance r from a randomly chosen entry hole divided by the number of entry holes and attack density. To remove the effects of the cut-ends, the areas within 20 cm from both cut-ends were removed. Fifteen logs with 20 or more entry holes were used for the analysis. We applied Ripley’s edge correction, which is a method to correct the K(r) value by weighted value defined as the proportion of a circumference within the study area in the circumference of a circle centered at one point and passing through another point [r) = πr2 [r) values with Ripley’s edge correction were estimated from 10,000 simulations. The median of expected K(r) values with Ripley’s edge correction under the CSR were also estimated from 10,000 simulations by randomly generating entry holes. Since the gallery system of P. proximus consists of 2–3 egg galleries of 3–7 cm, which are generally oriented horizontally [r) function up to r = 15 cm. When the 95% confidence envelopes of observed K(r) values were larger or smaller than the median of the expected K(r) values under the CSR, the distribution pattern of entry holes was statistically significantly aggregated or uniform at the distance r, respectively.To determine the distribution pattern of entry holes, we carried out a K( R 3.6.1  with the R 3.6.1 . The K(ress data . K(r) iser point –36. If ar) = πr2 ,34,35. Tzontally , the horAbies species. The number of entry holes was the response variable, surface characteristics  was an explanatory variable, the area of each surface characteristic was an offset term, and the log was a random effect. P-values were calculated by Wald chi-square tests and corrected using the Holm-Bonferroni method.To determine the effects of bark texture on the number of entry holes, we used generalized linear mixed models (GLMMs) with a Poisson distribution and a logarithm link, separately for each 12(r) function analysis . The K12(r) function is a generalization of the K(r) function to a bivariate point process [12(r) is defined as the expected number of entry holes (= 1) within distance r from the boundary of a randomly chosen rough bark (= 2) divided by the number of rough bark and the attack density. Entry holes located in rough bark were considered to have a distance of 0. To remove the effects of the cut-ends, the areas within 20 cm from both cut-ends were removed from the analyses, while the rough bark surfaces around knots located in the remaining areas even a bit were used for the analyses. Fifteen logs with 20 or more entry holes were used for the analysis. We applied toroidal edge correction, where the original part was duplicated and the edge on one side was thought of as being wrapped around to the opposite edge [12(r) values at 0.5-cm intervals up to 15 cm using ArcGIS and R. The 95% confidence envelopes of the observed K12(r) value with toroidal edge correction was estimated from 10,000 simulations. The median of expected K12(r) values with toroidal edge correction under the CSR was also estimated from 10,000 simulations by randomly sampling the same number of knots as the observed number with replacement. When the 95% confidence envelopes of the observed K12(r) values were larger or smaller than the median of the expected K12(r) values under the CSR, the spatial association (aggregation/segregation) between the entry holes and rough surface was statistically significant at a distance r.To determine the spatial association between entry holes and the rough surface around knots, we carried out a K process . K12(r) ite edge ,36. We mTo determine if there was a significant difference in phloem thickness under the rough bark around the knots relative to phloem thickness under the remaining bark, we used the Wilcoxon rank-sum test.The number of entry holes of each log ranged from 1 to 94 –S5 Figs,2, while that on the remaining parts ranged from 0 to 3.16 / dm2 .Attack density on the rough barks ranged from 0 to 11.19 / dm16 / dm2 . The rouOf the 15 logs, which had 20 or more entry holes, 14 logs revealed that entry holes had significantly aggregated spatial association with rough bark in the range of 5.5 cm, while one log did not reveal significant spatial association with rough bark .Rough bark around knots had significantly thicker phloem than the remaining parts of bark  values revealed that the distribution of entry holes was significantly aggregated within 5.5 cm from rough bark around knots in most cases. The GLMMs also revealed that rough bark had significantly more entry holes than the remaining bark. These results indicate that the bark surface texture affects the selection of the entry point of P. proximus. These also suggest that the increase in attack density on the rough bark around knots may result in the aggregated distribution of entry holes.However, the distribution pattern of entry holes was aggregated within 4.0 cm when the attack density was higher. The KIps typographus revealed a higher attack density around knots than that on the remaining part of the bark [Dendroctonus ponderosae by reducing their ability to grip a tree surface, resulting in a higher attack density on rough barks such as cracks, flakes, and crenulations. We revealed that rough bark around knots has thicker phloem than the remaining barks. Phloem thickness also plays a strong role in reproductive success and larval performance [P. proximus may preferentially attack rough bark to reproduce in the thicker phloem under the rough bark surface. To determine whether P. proximus preferentially attacks rough bark or avoids smooth bark, choice tests should be conducted.Previous studies have demonstrated that bark texture plays a strong role in the choice of attack location ,9,39. Ththe bark  suggesteformance –42. Our A higher attack density results in severe intraspecific competition for nutritious phloem among larvae and low reproductive success –3. The hS1 FigThe letters and numeric indicate log ID. Bar = 10 cm.(PDF)Click here for additional data file.S2 FigThe letters and numeric indicate log ID. Bar = 10 cm.(PDF)Click here for additional data file.S3 FigThe letters and numeric indicate log ID. Bar = 10 cm.(PDF)Click here for additional data file.S4 FigThe letters and numeric indicate log ID. Bar = 10 cm.(PDF)Click here for additional data file.S5 FigThe letters and numeric indicate log ID. Bar = 10 cm.(PDF)Click here for additional data file.


Beilstein Journal of Organic Chemistry that I have had the pleasure to edit, and these themed issues have gone progressively from strength to strength ever since the first was assembled in 2008. In this issue 45 papers are collected, a record for a thematic issue in the Journal, and there is no doubt that the Beilstein Journal of Organic Chemistry has become a settled home for papers in contemporary organo-fluorine chemistry. Authors have presented their recent findings or, in some cases, reviewed important areas, from bioactives to materials, and collectively the papers highlight the vibrancy and range of activity associated with this theme. Organo-fluorine chemistry merits our attention as an important discipline within contemporary organic chemistry contributing in a unique way to innovation and performance in the development of functional molecules.This is the fifth thematic issue on organo-fluorine chemistry in the This issue will be historically memorable as all of the manuscripts were submitted through the covid-19 virus lockdown period which impacted universities and research laboratories across the world. As Guest Editor I am indebted to all of the authors for maintaining their commitment to contribute through what was a particularly difficult and unusual period of professional life.Beilstein Journal of Organic Chemistry for their continuous attention to detail and the quality of the final published copy. The papers appear as contributions to the Journal in the normal sense, but they are also conveniently collected here as a single document.I thank the referees who gave of their time, and also the outstanding level of professionalism of the Editorial and Production Team at the David O’HaganSt Andrews, Scotland, February 2021


We report our experience regarding the profile and screening process of potential recipients (R) and their live donors (D) in our Uterus transplantation (UTx) trial from 2014 to 2020. The initial screening was performed using medical questionnaires and consultations. The second step of the screening consisted of two individual interviews with an independent multidisciplinary committee. Then, a complete medical, biological and imaging assessment of the directed living D, the R, and her partner was performed over a two-day hospitalization. A total of 239 women contacted our department: 165 potentials R and 74 potentials D. During the first step of screening, 141 R and 45 D were excluded. Only 12 R/D pairs were pursued. During inclusion, 10 R/D pairs were excluded. One R/D pair is still under evaluation. Finally, only 1 R/D pair was definitively included (0.6%), which led us to perform the first French UTx in March 2019 with a successful graft. The primary limiting factors of inclusion were due to very strict criteria and difficulty of having a suitable directed living D. The International Society of UTx (ISUTx) guidelines based on worldwide results of trials can help ease our inclusion criteria in the future while remaining safe for patients. Absolute Uterine Factor infertility (AUFI) affects approximately 1 in 500 women . AcquireAs part of the preinclusion phase of our UTx clinical trial (n° NCT03689842), we considered patients with AUFI who had independently contacted our institution. Starting in October 2014, we queried whether they would be interested in participating in our study. We collected data from this date until March 2020. Our trial is a prospective, open-label, non-randomized, single center study conducted in 10 patients with an MRKH syndrome or having suffered a hysterectomy (with no radiotherapy) for cervical cancer (stage IA FIGO 2018) in remission for ≥5 years, benefiting from a directed LD. There was no advertising or recruiting for the clinical trial. This study was approved by our institutional review board (CEROG 2018-GYN-1202) in January 2019.One hundred and sixty-five potential recipients interested in the trial contacted us with letters, dedicated email, or through direct consultation with our referring investigator. A medical heath history questionnaire was sent to all patients who didn’t have an appointment in order to evaluate if they could go further in the trial. We received 9 letters from potential recipients, and 64 patients contacted us only by mail. In total, 92 potential recipients and their partners had a consultation with our referring investigator (gynecologist). In addition, 45 potential donors had a consultation. The following data were collected from the potential recipient: age, residency, language, body mass index (BMI), cause of AUFI, directed donor, parental, marital, smoking and obstetrical status, medical and surgical history, medications, and partner’s opinion about UTx. In case of MRKH, any possible associated malformation and vaginal reconstruction were noted. The following data were collected for the directed donors: relationship status with the recipient, age, BMI, obstetrical history, smoking status, medical and surgical history, medications, and ABO compatibility with the recipient. If the recipient, the recipient’s partner, and their directed donor did not have exclusion criteria, they received medical information about the project. This included the experimental nature of the program, a description of the trial plan with all the pre-, intra and post-transplantation procedures, risks incurred, research advances in terms of transplantation, and the worldwide experience. Ultimately, the donors and recipients had a clinical check up with blood pressure, heart rate, abdominal, and gynecological examination conducted by the referring investigator.Donor, recipient, and their partner were interviewed individually after a 3-month reflection period by the Foch Hospital uterine transplantation multidisciplinary independent committee  The donor/recipient pair was informed of the lack of financial cost, as the study sponsor would be responsible for all costs related to the surgical procedure, transport, housing, and loss of earning during work interruption. During the first interview, the consent form and the information note were given and explained to the donor, recipient, and their partners. They were informed by the committee of the procedure, risks of complications, or failure in each step of UTx. They had an individual interview with the psychologist to evaluate their ability to participate in the UTx trial and undue familial pressure, especially for donors. After the first and the second interview (following a 3-month reflection period), the multidisciplinary uterine transplantation committee decided on the patient’s eligibility. Finally, the consent dated and signed by the donor, the recipient and her partner were collected by the investigator after the approval of the multidisciplinary committee.A medical, psychological, biological, and imaging assessment of the directed living donor, the recipient and her partner were performed during a 2-day hospitalization in order to confirm the eligibility.During the first day, a general assessment , anesthesia consultation, and psychological consultation were performed. Based on the results, the inclusion process could be stopped and the couple excluded.During the second day, a general assessment (imaging) and a psychological consultation were performed. Depending on the results, the inclusion process could be stopped and the couple excluded. In case of inclusion, a donor was interviewed individually by the biomedicine agency Living Donor Committee. After the living donor committee approval, the donor had to express her consent to the president of the Court of first Instance.Obtaining 10 cryopreserved embryos at the blastocyst stage from the recipient and partners gametes was a requirement for including the recipient and their directed donor. Seven days before surgery, a shorter medical, psychological, biological, and imaging assessment was performed again in order to check the inclusion criteria. The screening and inclusion criteria are summarized in Quantitative data were expressed as a mean (range) and qualitative data were expressed as percentages.A total of 239 women contacted our department for our UTx trial from October 2014 to March 2020: 165 potential recipients and 74 potential donors. The characteristics of potential recipients and donors are summarized in It is important to select the best R and D candidates in order to obtain a functional graft and, at term, a pregnancy with the birth of a healthy child with a minimum of risk for recipients, donors, and resulting children. Moreover, this is a new procedure still in the experimental stage and the precautionary principle must be widely applied. We found that, out of 165 UTx candidates with a directed donor, only one was able to integrate our trial; this is the lowest inclusion rate in reported trials ,27. We cIn our trial, we only used LD directed to the recipients. These had emotional or genetic relationships with the R on the model of kidney transplantations. Most often, they were the mothers, as in the majority of UTx trials. In 72% of cases, the directed donor failed to fill the inclusion criteria at the first step of the screening. In order to increase the number of potential compatible donors, the Baylor’s team included altruistic donors . Non-dirThe risk of infection is high mainly during the first six months after a solid organ transplantation because of the immunosuppressive therapy and during pregnancy due to the physiologic immunomodulation. Explantations of the grafted uterus for uterine abscess, HSV, and candida infections have been reported ,15,17. AUTx from deceased donors, still in its infancy, is an interesting alternative to avoid LD complications ,43. It cUp to now, R were mostly MRKH, the congenital AUFI associated with shortened vagina. A sufficient vaginal length is necessary to perform a good anastomosis with the graft and decrease the risk of vaginal stenosis after UTx . VaginalA sufficient but thoughtful screening process of living donors and recipients is essential and should aim both to assure donor/recipient safety and to provide good quality grafts. The risk/benefit ratio and evolutions of UTx will be determinant to define which R and D could be included in the future.


The degree of molecular order (DMO) and degree of double helix (DDH) were significantly (p < 0.05) reduced from 1.35 to 1.01 and 1.38 to 0.98 respectively. X-ray diffraction (XRD) indicated the consecutive decrease in relative crystallinity (RC) while RVA analysis showed that peak and final viscosities were decreased significantly (p < 0.05). However, digestion kinetics indicated that degree of gelatinization increased the access of enzymes. As starch was partially gelatinized it yielded significantly lower glycemic index but no significant (p > 0.05) change in starch digestibility was observed after 70 °C. Hence, 70 °C can be considered as the critical hydrothermal treatment temperature in mung bean starch. Pearson’s correlation analysis indicated that controlled hydrothermal treatment had negative effect on the DMO, DDH, RC and the granular damage increased vulnerability of mung bean starch to digestion. These findings gave insight into sequential changes in the structure and digestibility occurring during gelatinization process due to hydrothermal treatment. Controlled gelatinization in mung beans at 70 °C is useful and must be employed to produce the foods with lower starch digestibility.The changes in structure and digestion properties of mung bean starch due to hydrothermal treatment at various controlled temperatures were investigated. Results showed the increase in onset temperature (T Vigna radiata L.) is a leguminous crop excessively cultivated in South-Asia and it has potential health benefits [−1 on dry weight basis which is extracted by employing different processing methods [Mung bean  was purchased from Gucheng Sandou Food Technology Development Co. Ltd., . Salivary α-amylase , pepsin , porcine pancreatic enzymes , amyloglucosidase  were used for starch digestion. D-Glucose  was used for the glucose measurements. All other reagents used in experiments were of the analytical grade.v/w) extracted mung bean starch were prepared at room temperature in polypropylene bags. Starch samples were hydrothermally treated by following the method of Wang et al. [Starch was extracted from mung beans by previous method of extraction . Aqueousg et al. . The bagThermal properties of the treated starch samples were analyzed using a differential scanning calorimeter  equipped with thermal analysis data station following the method of Yao et al. . BrieflygelH. is the enthalpy changes in gelatinized starch sample and ∆natH. represents the enthalpy changes in the native starch.Degree of gelatinization (DG) was further calculated by using the previously used formula by . It tell−1 and holding time at 95 °C was 5 min. Then it was cooled down to 50 °C with a decrease of 6 °C min−1 and held there for 2 min. The studied parameters included: peak viscosity (PV), trough viscosity (TV), breakdown, final viscosity (FV), and setback.The pasting properties were determined using RVA  following the method of Guo et al. . BrieflyA scanning electron microscope  was used to study morphological changes in the structure of gelatinized mung bean starch granules. Dried starch samples were mounted on SEM stub and pasted on double-sided adhesive tape and sputter-coated with gold in a vacuum evaporator. All the specimens were studied under the accelerating voltage transmission of 10 kV. Images were taken at 500× magnification power (10 µm) and 4500× magnification power (100 µm) for single and grouped starch granules respectively.V/V) and observed under the polarized light microscope at 40× to check the birefringence of the starch.A polarization microscope  equipped with a SPOT insight camera was used to analyze the birefringence in starch granules at different degrees of gelatinization in bean starch. The analysis was performed following the previously used method of Li et al. . Mung beλ) of 0.1542 nm. The starch powder was tightly packed in a rectangular glass cell and scanned over the range of 4° to 40° with a function of 2θ. The scanning speed during the analysis was 2 °C min−1 and scanning step was 0.033°. The relative crystallinity was quantitatively measured by following the method of Yao et al. [A diffractometer  was used to study the crystallinity of dried pre-gelatinized starch samples with moisture content around 10%. All the measurements were performed at 40 kV and 40 mA with Cu-Kα monochromatized radiation with a wavelength  and degree of short-range molecular order (DMO) by using the described method of Xu et al. [Fourier transform infrared (FTIR) spectra were obtained by using a Fourier transform infrared spectrometer  following the method of . The pow2, 200 mM CaCl2). Then it was placed in shaking incubator at 37 °C for 15 min to attain the temperature. Then a freshly prepared enzyme solution of porcine pancreatic amylase (25 µg) and amyloglucosidase (AMG) was added to the starch suspension. Starch digesta were collected at the regular time intervals . Enzyme reaction was stopped by the addition of 95% ethanol in the aliquots collected at different time intervals. Undigested starch was removed by centrifugation at 2000× g for 5 min at 4 °C. The starch digestibility was calculated by measuring the amount of released glucose by using the kit method (Megazyme Glucose Format Kit). The digestograms were fitted to first-order kinetics as previously described method by Edward et al. [tC indicates the concentration of digested starch at incubation time ∞t, C is the estimated digestion at infinite endpoint of the reaction and k is the digestion rate coefficient which was measured using a logarithm of slope (LOS) plot analysis using the natural logarithmic form of Equation (2) expressed as Equation (3):dC/dt) can be plotted against digestion time t, which has linear relationship between the slope of −k and time t. However, ∞C was calculated by the intercept of the equation and slope k.Extracted native and heat-treated starch samples (50 mg) were dispersed separately in 2 mL of deionized water and incubated with 7.5 mL of sodium acetate buffer  were calculated. Analysis of variance (ANOVA) was performed by least significance difference (LSD) using SPSS V. 22.0 windows software . Duncan’s multiple range test was applied and the level of significance was set at Differential scanning colorimetry is an effective tool to measure the endothermic transitions relating to the dissociation of amylopectin and double helices from semi-crystalline to amorphous arrangement. The endothermic values of controlled treated mung bean starch are shown in o), Peak (Tp), and Conclusion temperatures (Tc) were increased which can be attributed to the melting of less stable crystallites during the pre-treatment. Hence, higher energy input was required for the dissociation of larger starch crystallites. Similar findings were previously reported about DG and thermal properties by Wang et al. [With the increase in degree of gelatinization (DG), Onset  and enthalpy of retrogradation (ΔrH) were decreased from 14.07 to 6.77 J·g−1 and 10.06 to 6.68 J·g−1, respectively, with increasing end temperature of pre-heat treatment. This was attributed to the melting of less stable crystallites initially and later more energy input was required for dissociation of larger crystallites [Enthalpy changes are due to combination of endothermic disruption process of ordered structure and the exothermic process of granular swelling . Contrartallites .p > 0.05) and progressively with increase in thermal pre-treatment temperature from 6613.50 mPa.s to 2015.50 mPa.s from native starch at 100 °C. This can be attributed to maximum damage of starch granules, which was significantly (p > 0.05) reduced with the increase of temperature. The rotating paddle was used to measure the resistance of starch during the heating phase which was termed trough viscosity. The trough viscosity was reduced from 3455.50 mPa.s in native starch to 1835.50 mPa.s for maximum gelatinized samples (100 °C) due to loss of inherent capacity of starch granules as surface structure is damaged and water moves inside easily. Hoover et al. (2011) described pasting to be primarily dependent upon swelling characteristics of granules, leaching of granules and chain length distributions (CLD’s) of amylopectin [The pasting properties of native and cooked starch samples determined by RVA are shown in lopectin .The point where starch granules are ruptured is termed as breakdown and was calculated from peak and trough viscosity . For natThe scanning electron micrographs of cooked starch at different temperatures i.e., 50 °C, 60 °C, 70 °C, 80 °C, 90 °C, and 100 °C are shown in the X-ray diffraction profiles of gelatinized mung bean starch with relative crystallinity (%) at various temperatures are shown in −1 and 995/1022 cm−1 can be related to the degree of short-range molecular order and degree of long-range double helix, respectively [−1 shown as 3390 cm−1 in −1. −1. The short-range ordered structure was still present at the maximum heat treatment temperature (100 °C). The results were comparable with Xu et al. [p > 0.05) between the native and autoclaved pea and navy bean samples, which was attributed to the uncoiling of the double-helical structures of starch granules.The infrared spectra have been displayed in ectively . The shok values were increased 2.5-fold between native and the gelatinized sample. The digestion extent (∞)C at the endpoint of reaction varied significantly (p < 0.05) from native (22.69 ± 0.50%) to 78.99 ± 0.10% for samples gelatinized at temperatures up to 80 °C. However, no significant difference was observed when the starch was completely gelatinized in heat-treated samples at 80 °C, 90 °C, and 100 °C. Similar results were reported by Wang et al. [In vitro digestion kinetics of gelatinized mung starch to different degree of gelatinization are given in g et al. , where tg et al. . The accg et al. . The relg et al. . Keepingg et al. .o) was highly positively correlated with all other DSC parameters  but degree of gelatinization (DG) was found to be negatively correlated (p < 0.01). This indicates that the more starch is pre-treated or gelatinized, the lesser will be its thermal viability. The digestion extent (∞)C and the value of k were found strongly correlated (0.898 **) and (0.997 **) with the degree of gelatinization (DG) respectively. This suggests that the pre-gelatinized starch tend to digest more easily due to damage of granular structure and easier initial binding of amylases to the starch.The values of Pearson’s correlation analysis are shown in ∞).C The controlled thermal treatment of starch lead to the decrease in the structural orders and increase in the digestion extent of the starch. These findings were found contrary to the results of Li et al. (2017) [gH and ∆rH).The structural parameters such as degree of molecular order (DMO), degree of double helix (DDH) and relative crystallinity were found to be strongly negatively correlated with the digestion extent (. (2017) , where nControlled gelatinization of Mung bean starch was carried out at different temperatures and the transitional changes in the digestibility and structure were observed. The structure-digestion relationship was observed by Differential scanning colorimetry (DSC), Scanning electron microscopy and short-range molecular orders. The results indicated that 70 °C was the ideal gelatinization temperature for lower glycemic index but at further gelatinization, the structure was disrupted and in-vitro digestibility was maximum. Mung bean products especially noodles can be cooked at 70 °C for slightly longer time to reduce the glycemic index. Moreover, mung bean composites can be prepared with other key cereals to reduce the glycemic index and insulinemic responses. Correlation analysis indicated that controlled gelatinization treatment had negative effect on the amorphous and crystalline orders of starch structure and the granular damage increased the vulnerability of mung bean starch to digestion. It was found that controlled gelatinization was useful to control the digestibility of starch.Various other methods of thermal treatments such as microwave and roasting can be used to control the degree of gelatinization in the future to get the lower starch damage and lower starch digestibility.


However, fatigue did not cause a statistically significant reduction in fracture resistance in both LD and HL groups (p > 0.05). Copings fractured in the LD crowns only and the fracture path was different in both LD and HL groups. According to the results, it was concluded that milled veneer implant-supported hybrid-abutment crowns exhibit significantly higher fracture resistance, and better withstand clinical masticatory loads in the posterior region compared to the hand-layered technique. Also, fatigue application and artificial aging caused no significant strength reduction in both techniques. Clinical significance: Different veneering techniques and materials (hand-layering or milling) act differently to clinical forces and environment and may be prone to early chipping during service. Therefore, practitioners are urged to consider the appropriate veneering protocol for posterior implant-supported hybrid-abutment restorations.The objective of this study is to investigate the influence of veneering technique (hand-layering vs. milling) on the fracture resistance of bi-layer implant-supported zirconia-based hybrid-abutment crowns. Mandibular molar copings were anatomically designed and milled. Copings were then veneered by hand-layering (HL) (n = 20) and milling using the Cad-On technique (LD) (n = 20). Crowns were cemented to zirconia hybrid-abutments. Ten samples of each group acted as a control while the remaining ten samples were subjected to fatigue in a chewing simulator. Crowns were loaded between 50 and 100 N for 1.2 million cycles under simultaneous temperature fluctuation between 5 and 55 °C. Crowns were then subjected to static load a to fracture test. Data were statistically analysed using the one-way ANOVA. Randomly selected crowns from each group were observed under scanning electron microscopy to view fractured surfaces. Study results indicate that during fatigue, LD crowns had a 100% survival rate; while HL crowns had a 50% failure rate. Fracture resistance of LD crowns was statistically significantly higher than that of HL crowns at the baseline and after fatigue ( Being the toughest of all dental ceramics , zirconiCopings of bi-layer zirconia crowns can be veneered using multiple techniques such as conventional hand-layering technique, pressing technique or milling technique using the computer aided design/computer aided manufacturing (CAD/CAM) technology. In the conventional hand-layering technique, a build-up of dentin and enamel porcelain is done directly on the coping. However, in the pressing technique, the veneer is pressed on the coping using an ingot and a special press furnace; this provides superior strength  and anatSatisfactory bonding between the coping and the veneer material can enhance fracture strength and is the key to bi-layered restoration success ,17. It iBeing more aesthetically pleasing ,25, highA new approach to achieve aesthetics and strength in implant dentistry is the hybrid-abutment crown approach ,30. It iTo date, no information is available about the influence of the veneering technique used in the bi-layer structures on the fracture resistance of such hybrid-abutment crowns. Therefore, this study aimed to evaluate the fracture resistance and post fatigue fracture load of zirconia copings veneered with CAD/CAM milled lithium disilicate structures compared to hand-layered veneered copings when cemented to hybrid-abutments and supported by implants. We hypothesised that implant-supported hybrid-abutment crowns veneered with milled lithium disilicate ceramic would exhibit similar survivability as well as fracture loads to failure compared to hybrid-abutment crowns veneered by hand-layered nano-fluorapatite ceramic. We also hypothesised that fatigue testing in a simulated oral environment would not significantly affect fracture loads for crowns made in either technique.® CAD-On) (n = 20) by joining a milled lithium disilicate veneering structure (LD) to the zirconia coping, and hand-layering technique (n = 20) by building up dentin and enamel ceramic on the zirconia (HL). Materials and components used in this study are listed in For the purpose of this study, hybrid-abutments  were use® select, Wieland Dental, Lindenstraße, Germany) to mill zirconia abutments (n = 40) and zirconia copings (n = 40). Zirconia structures were then sintered in the recommended Programat S1® furnace  and left to slowly cool. CAD-On crowns were designed using the split-file technique ; anatomical zirconia coping was designed first and followed by designing the lithium disilicate veneering structure. The zirconia abutments and the CAD-On design files were then transferred to a 5-axis milling machine , LD veneering structures (n = 20) were milled from the same CAD-On design file previously used to mill the zirconia abutments and copings. Afterwards, fitting was checked for: (a) zirconia abutments to the Ti-Base, (b) zirconia copings to zirconia abutments and (c) LD veneering structures on the zirconia copings. Afterwards, IPS e.max® Crystall./Connect capsule was mixed using the Ivomix  and evenly distributed on the occlusal aspect of the zirconia copings as well as on the fitting surface of the LD veneering structures. Both copings and veneering structures were then joined together using Crystall./Connect and any excess material was removed from the circular fusion joint. To verify the correct join between the coping and the veneering structure, all LD crowns were checked for occlusion in an articulator. The recommended Fusion/Crystallisation firing of the LD crowns was conducted in a Programat EP 3010® furnace . Finally, LD crowns were glazed and recommended glazing firing was also conducted in the same Programat 3010® furnace.In a wet milling machine , hand layering of the zirconia copings was done by a dental technician with 20 years of experience. The silicone index of an LD crown was taken and used during the porcelain build-up process of dentin and enamel porcelain of the HL crowns; to ensure standardisation of both anatomical shape and thickness of the layering porcelain in both LD and HL crowns. Then, all crowns were glazed as per the manufacturer recommendation using the IPS e.max Ceram glaze. Firing of the dentin porcelain, enamel porcelain and glazing was conducted as per manufacturer recommendation in a Programat 3010® furnace .Using IPS e.max® Hybrid Abutment, Ivoclar Vivadent, Schaan, Liechtenstein) as per the manufacturer instructions.To cement the crowns to the Ti-Base, the Ti-Base access hole was first filled with a temporary restorative material , and zirconia abutments were then luted to the Ti-Base using a self-curing dental luting composite  as per manufacturer instructions. Specimens were stored in distilled water at 37 °C for a minimum of 7 days prior to testing [Twenty-four hours later, both LD and HL crowns were adhesively luted to the zirconia abutments using Multilink testing .Both LD (n = 20) and HL (n = 20) groups were then divided into two subgroups (n = 10) according to the testing to be conducted; (1) control group (non-fatigued) crowns to be subjected to static load to fracture (SLF) in a universal testing machine, (2) test group (fatigued) to undergo thermal cycling mechanical loading in a chewing simulator prior to the SLF test.To prepare samples for fatigue testing  in the cUsing a 6-mm diameter stainless steel spherical indenter and a loading frequency of 1.2 Hz, crowns were loaded for 1,200,000 cycles in a chewing simulator to simulate 5 years of clinical service ,41,42. TBoth control groups  were subjected to compressive SLF loading test  until failure. The compressive loading was applied on the crowns at three points  , to check the fractured surface condition and highlight any differences between the groups (if present). Sputter coating of the crown was done with gold  to a thickness of approximately 10 µm prior to imaging.p value equal to or less than 0.05 was set to indicate statistical significance.Statistical analysis of the data was conducted using the SPSS software . Skewness and Kurtosis tests and P-P plots were used to check the normality of data distribution. One-Way Analyses of Variance (ANOVA) was used to compare means and to evaluate statistical significance between all study groups. A post-hoc assessment was performed using the Tukey HSD test. T-test was also conducted to analyse data of the HL crowns of the fatigued group (survived vs. failed). A No implant fractures occurred during fatigue in the chewing simulator in both LD and HL groups. Also, no failure occurred for the LD crowns during chewing simulation. Nevertheless, HL veneered crowns failed by means of chipping under the indenter contact point during chewing simulation and at a different number of cycles; two crowns failed during the first stage of the loading protocol under 50 N; at the end of 250,000 cycles. Also, three crowns failed during the second stage of the loading protocol less than 100 N; one failed at 418,000 cycles and the other two failed at the end of the 500,000 cycles. Wear facets on the occlusal contact of the indenter were evident in both LD and the survived HL crowns .The ultimate fracture load values (standard deviation) recorded in newtons for un-fatigued crowns were as follows:Crowns veneered with milled lithium disilicate (n = 10):Hand-layered veneered crowns (n = 10):On the other hand, the ultimate fracture loads values (standard deviation) recorded in newtons fatigued crowns were as follows:Crowns veneered with milled lithium disilicate (n = 10):Hand-layered veneered crowns (n = 10):p ≤ 0.05) than HL crowns. However, there was no statistically significant difference between the control and the fatigued crowns in both groups.Mean and standard deviation of fracture loads in Newtons (N) for all study groups are presented in Upon conducting the SLF test, it was observed that crowns of HL subgroups had a cohesive fracture of the veneering on the lingual cusps, which extended to the coping-ceramic interface with no fracture of the coping. On the contrary, both veneer structure and copings fractured in LD subgroups. In addition, no ceramic abutments, Ti-Base or implants were fractured and no screw loosening occurred in any of the groups.Representative SEM images of fractured surfaces are presented in This study looked at bi-layer zirconia-based implant-supported hybrid-abutment crowns in the posterior region. Our results showed that zirconia-based copings for hybrid-abutment crowns veneered with a lithium disilicate veneer had significantly higher fracture load to failure compared to the hand-layered veneer. Also, fatigue did not cause significant reduction in fracture resistance for crowns veneered in either technique with milled lithium disilicate veneers resulting in 100% survival rate. Nonetheless, some hand-layered veneered crowns failed during fatigue testing by means of chipping. These findings reject our hypothesis that hybrid-abutment crowns made of zirconia copings and veneered with milled lithium disilicate exhibit similar fracture loads and survivability compared to crowns with hand-layered veneers. However, the hypothesis that fatigue testing will have no significant effect on the fracture load of hybrid-abutment crowns veneered in both techniques was accepted. Past experimental or clinical studies provide no semblance to the current work. Thus, results of this study may be hard to directly compare to other studies conducted on hybrid-abutment crowns.Fracture resistance of crowns with CAD/CAM lithium disilicate veneer was significantly higher than that of the hand-layered veneers. Studies on tooth-supported crowns reported the same results with CAD/CAM veneered zirconia coping, displaying significantly higher fracture loads compared to the hand-layered veneers ,47,48. FFive crowns of the hand-layer veneered group suffered chipping of the veneer during fatigue testing on the cusp where the indenter was loaded. The high veneer chipping rate of hand-layered zirconia-based restorations was also reported in a previous study which compared both veneering methods in tooth-supported crowns . Kassem Such significant difference in fracture load values between both veneering techniques might be due to differences in both veneering materials and fabrication techniques used; while a lithium disilicate veneer has a flexural strength of 360 MPa, the veneering ceramic used in the hand-layering technique has a flexural strength of 90 MPa making the later more prone to failure at low loads during mastication . Also, lIn the oral cavity, chipping usually occurs after exposure to a localised load on one or more cusp during mastication while undergoing aging at the same time . For theAgreeing with previous studies ,35, neitWithin the limitation of the current study, it was concluded that veneering hybrid-abutment zirconia-based crowns using a milled lithium disilicate structure creates a crown with high fracture resistance properties, and may result in highly clinically reliable restorations. Also, we concluded that crowns veneered using the conventional hand-layering technique are likely to fail by chipping of the veneering layer early during clinical service, which makes them unfavourable and less cost effective for both the clinician and patient. Furthermore, it was concluded that the combination of zirconia abutment to a Ti-Base (hybrid-abutment) has an adhesive bond that is unlikely to fail during clinical service.Results of this study are only an indication of the hybrid implant abutment system’s behaviour in an in-vitro environment; we suggest that short and long-term clinical trials are necessary to assess implant-supported hybrid-abutment crowns veneered in both veneering techniques.


Domino-liver transplantation represents a rare chance to expand the donor liver pool. Fear of putting both donor and recipient at disadvantage has meant that the procedure has not been applied universally. A modification of the original technique which allows both safe procurement of the graft as well as safe implantation of the reconstructed graft in the domino-graft recipient using a 180° rotated, adequately trimmed, free iliaco-caval venous graft is described in detail. Domino  liver transplantation (DLT) represents an opportunity to expand the liver allograft pool. The first Swedish experiences with liver transplantation (LT) for hereditary transthyretin amyloidosis, also known as familial amyloidotic polyneuropathy (FAP), the high incidence of this disease in Portugal and the structural normality (except for the production of the mutant TTR) of such livers, led to the development of DLT –3. The cabove the diaphragmatic ring to assure a sufficiently long supra-hepatic IVC cuff, and the systematic use of the veno-venous bypass (VVB) to overcome hemodynamic instability caused by IVC clamping in neurologically dysregulated FAP patients, clearly disadvantaged the domino-donor compared to the ‘classical’ liver transplant patient [Over the years, technical developments, routine introduction of inferior vena cava (IVC) sparing hepatectomy techniques, and accumulated experiences in centers specializing in liver-based metabolic diseases overruled these concerns –20. The  patient . Ten yea patient , 21–23.  patient –27. A fuDLT is performed only following informed consent obtained from both domino-donor and domino-recipient patients after a detailed explanation of both donor and recipient transplantation procedures. DLT was approved by the ethical review board of the Université Catholique de Louvain.The liver is rolled off from the IVC and prepared from below upwards after ligation of all hepatic veins draining segment I and of any larger right inferior (Makuuchi) veins. The right hepatic vein (RHV) and the cuff of middle (MHV) and left hepatic (LHV) veins are isolated separately. Suture ligation and transection of the left phrenic vein allow lengthening of the MHV–LHV cuff. Grasping the tendinous ring of the diaphragm with a Babcock clamp makes the dissection of the supra-hepatic IVC easier, because it better exposes the dissection plane. The diaphragmatic portion of the supra-hepatic IVC and the para- and retrocaval areas are left intact. The dissection of the hilar structures is guided by the pre-transplant angio-CT scan, which is done in all domino-donors and domino-recipients to get accurate information about existing vascular variations. After dissection of the proper, common, and gastroduodenal arteries, these arteries are bull-dog clamped and the proper hepatic artery is divided at the bifurcation of gastroduodenal and common hepatic arteries in such a way that a small Carrel patch can be obtained. Small (left) accessory arteries are ligated. In case of separate right and left hepatic arteries, these will need to be implanted separately. The portal venous trunk is divided in its middle and the bile duct is transected just above the level of the cystic duct. The RHV is transected vertically using an endovascular stapler . Precise stapler application close to the liver parenchyma permits a rapid, safe, and tight transection of the RHV without narrowing the IVC.Moreover, this maneuver avoids bleeding from the parenchymal side when extensively mobilizing the liver and allows the liver to be rolled off further from the IVC, helping in the safe isolation of the MHV and LHV. Broad experience with this type of vascular closure in classical LT and living donor LT revealed that the removal of a precisely placed stapler line allows obtaining a clean, rectilinear, and longer RHV stump. Indeed, in case the RHV is closed using a running suture, the varying and thicker amounts of a vascular wall comprised in such suture frequently oblige to resect a larger vessel rim. Consequently, the RHV may be shorter.The inferior RHV, present once in these series, was simply suture-ligated even if larger than 5 mm.The isolated M-LHV cuff is double clamped with a large right-angled vascular clamp. After scissor transection, the distal clamp is removed, and the cuff is closed using running sutures. The post-mortem allograft is implanted in the domino-donor using a large latero-lateral cavo-caval anastomosis under partial IVC clamping using the Lerut–Satinsky vascular clamp . Its prolonged vertical and horizontal part allows a safe lateral clamping of the IVC and so a comfortable suturing of donor and recipient IVCs. All other vascular and biliary sutures are done as usual .The exsanguinated liver is flushed with HTK or IGL solution through the cannulated short portal vein. The RHV stapler line is taken down and the liver gently massaged to allow rapid emptying of its blood content. Depending on the anatomical variation, LHV and MHV are together or separated. Venoplasty (if close to each other) or use of a small venous patch (if too far from each other) allow one single orifice to be created . The easThe diameters of the newly constructed supra-hepatic cuff and the M-LHV cuff of the domino-recipient will fit, thereby allowing a safe piggy-back implantation of the domino-graft. The length of the IVC interposition graft needs to be trimmed ‘in situ’ in such a way that kinking, and outflow obstruction are avoided. All anastomoses are performed using running polypropylene 5 to 7/0 sutures. This type of implantation can be done in the domino-graft recipient without total IVC clamping and so without VVB use.During the period February 2000–January 2017, 16 LT were performed for FAP. On three (18.7%) occasions, DLT was not possible due to extended intrahepatic and extrahepatic portal vein thrombosis (1 pat) and advanced fibrosis (2 pats) due to autoimmune cirrhosis and restrictive amyloidotic cardiomyopathy. One Fap liver was split for two small adults. Indications for DLT were hepatocellular cancer (7 pats), primary biliary cholangitis (3 pats), alcoholic cirrhosis with severe encephalopathy (3 pats), and neuro-endocrine metastases (1 pat). IVC-sparing hepatectomy without VVB use was performed in all LT procedures. Median warm ischemia time of the domino-graft was 43 min (range from 30 to 73). In one case, the hepatic artery had to be anastomosed to the splenic artery because of fibrotic changes of the hepatic artery caused by trans-arterial chemo-embolisations; in one right and left, hepatic arteries were anastomosed to similar, corresponding recipient arteries, and in another, the portal vein needed to be prolonged using a free iliac vein to overcome an extended portal vein thrombosis. The day-7 angio-CT scan of the patient in which a dominant inferior RHV was not implanted revealed a normal venous outflow of the right liver. In all domino-grafts, Doppler ultrasound revealed normal, triphasic, outflow patterns, and angio-CT showed a well-vascularized segment I. Two domino-donors presented a right pleural effusion. None of the domino-graft recipients presented after a median follow-up of 40 months (range 1–206) signs of disease transmission. One domino-liver recipient developed an ascitic decompensation 10 years post-DLT due to fibrotic changes of the neo-supra-hepatic cava cuff; a single balloon dilatation resolved this problem. In this case, a stored iliaco-caval graft from another donor had been used to reconstruct the venous outflow. Four domino-recipients died of recurrent cancer , two of cardiac complications (at one and six mo), one each of pulmonary fibrosis (at 152 mo), suicide (39 mo), and massive variceal bleeding (1 mo). There was no documented disease transmission in any of the domino-graft recipients.Domino LT represents a rare opportunity to enlarge the liver donor pool . The extThe technical simplification of DLT described here addresses the challenge of the outflow reconstruction, the avoidance of VVB use in both domino-donor and domino-recipient, and possible interference with thoracic and renal procurement procedures related to venous graft harvesting. The modified outflow reconstruction of the domino-graft should represent a standardized approach to use in domino-liver transplantation, aiming thereby at optimal use of these precious grafts.


The subunits are arranged as one or two 19S regulatory particle(s) capping a cylindrical catalytic 20S core particle . Each subunit is highly conserved from yeast to mammals, and proper function of the proteasome is crucial for survival of organisms . Spatiotemporal expression of specific subunits and their roles in regulating the proteasome activity is still not clearly understood.The 26S proteasome is one of the major proteolytic machineries in cells and is composed of nearly 33 different subunits . In C. elegans, loss or down regulation of a single proteasome subunit causes embryonic lethality, with the exception of three of the 19S RP subunits: RPN-10, RPN-12, and DSS-1 . While the roles of RPN-10 and DSS-1 were previously studied, more detailed investigation into the function of RPN-12 has not been performed. A heterozygous mutant containing a 952bp deletion of the coding region of rpn-12 was generated [rpn-12(av93)] using CRISPR/Cas9 genome editing technology. The heterozygous rpn-12 null mutant was then homozygosed and determined to be homozygous viable with apparent fertility issues. Only 36% of the rpn-12(av93) hermaphrodites produced progeny due to sperm production defects (this phenotype will be published elsewhere). Since efficient proteasome activity is important for the longevity of most organisms, we wanted to investigate whether rpn-12(av93) mutants may have an altered lifespan compared to wild type animals . To determine whether the lifespan of rpn-12(av93) mutant animals was affected, lifespan assays were conducted at 20°C and 25°C. The mean lifespan of rpn-12(av93) hermaphrodites (15.94 days) was not significantly different from wild type hermaphrodites (15.54 days) at 20°C. Interestingly, at 25°C the mean lifespan of rpn-12(av93) hermaphrodites (15.59 days) was increased compared to wild type animals (12.80 days). However, the maximum lifespan of both rpn-12(av93) and wild type hermaphrodites was not significantly different at 25°C. The lifespan analysis at 20°C was performed three times (cumulative wild type n=240 and mutant n=192) and at 25°C was performed twice (cumulative wild type n=185 and mutant n=135). Our data suggests that RPN-12 is not essential for viability and lifespan of C. elegans under normal conditions (20ºC), but that absence of RPN-12 can result in a significant increase in mean lifespan under heat stress (25ºC).RPN-12 is a subunit of the 19S regulatory particle (RP) of the 26S proteasome in rpn-12(av93) AG343 strain was generated via CRISPR/Cas9 genome editing technology following the direct delivery method developed by the Seydoux laboratory . Two crRNAs (5’- agccagaagatttttatggg – 3’and 5’- actcgaacaaatcgtttaac – 3’) were used to guide the Cas9 to make cuts at specific regions of either ends of the rpn-12 gene. An ssODN (5’-aaacattattggatttaagaaaatgtctgccgcccaaccggtgtctttcaagaactcaagcattgtattt-3’) was delivered as a template for the repair that results in a 952bp deletion that removes most of the first exon and then the entire second to last exons. Screening was performed using the co-conversion dpy-10 method . Two independent strains of the rpn-12 deletion were generated [rpn-12(av93) AG343 and rpn-12(ana10) WDC10] through the previously described method, and each strain outcrossed five times before experiments conducted. Both strains showed similar viability and fertility phenotypes, however only rpn-12(av93) was used for the lifespan experiments. Lifespan assays were conducted by placing age synchronized L4 larvae of rpn-12(av93) and wild type animals on MYOB plates seeded with OP50 (10-20 worms per plate). The worms were then passaged to new MYOB plates every two days. The number of dead hermaphrodites was scored every two days until all worms were dead and the percentage of surviving animals calculated. Missing worms were excluded from the statistical analysis.The rpn-12(av93)AG343 rpn-12(ana10)WDC10


The aim of the study was to assess the possibility of using polysaccharides: inulin and polydextrose in combination with steviol glycosides as sucrose substitutes in starch-based desserts with reduced sugar content and to determine their influence on the rheological properties of these desserts. The samples (starch-milk desserts) were prepared from native potato starch, milk, dye, flavouring agent, and sucrose. The sucrose was partially or completely substituted with steviol glycosides and inulin or polydextrose. The rheological evaluation of the desserts was performed by determining pasting characteristics, viscosity curves, creep and recovery curves and mechanical spectra. Substitution of sucrose with prebiotic polysaccharides modified the rheological characteristics of the starch-milk desserts to a degree depending on the type and level of the substituting agent. Inulin reduced the peak viscosity of starch-milk paste, while it had no effect on the final viscosity of the product, contrary to polydextrose, which increased value of the latter parameter. The desserts exhibited a non-Newtonian, shear-thinning flow behaviour. The use of inulin, in both the highest and the lowest concentrations, significantly changed the consistency coefficient and the flow index values, while such a phenomenon was not observed in the case of polydextrose. The desserts with inulin showed increased values of the storage modulus and reduced susceptibility to stress, manifesting strengthened viscoelastic structure. The results indicate that the both prebiotic polysaccharides can serve as substitutes for sucrose in desserts with reduced sugar contents. Starch, a natural biopolymer, is a reserve plant material. It is also a renewable raw material, commonly used in the food industry and other industrial branches . Starch Natural or modified starches are commonly used thickeners and texture creating agents in milk-based desserts ,12,13. TStevia rebaudiana Bert plant [It is well known that excessive consumption of sucrose promotes the development of diseases, such as obesity, diabetes and tooth decay. For this reason, many consumers are interested in low-sugar or sugar-free foods, including desserts . In the rt plant . Their crt plant ,15.Lactobacillus spp. and Bifidobacterium spp. in the colon to produce short chain fatty acids [In addition to the use of high-intensity sweeteners, reduction of the sucrose content in many food products also requires the use of bulking agents. One of the most important factors influencing consumer acceptance of a food product is its texture, which can be created or controlled with the use of food ingredients or additives. Such substances are, among others, inulin and polydextrose. Due to their texturing properties, low energy value and prebiotic properties, they are desirable food ingredients . Inulin ty acids . The lowty acids . In foodty acids . Polydexty acids . This poty acids . PolydexSince substituting of sucrose with small amount of intense sweetener results in modification of rheological characteristics of low- or reduced-sugar products, the aim of this study was to assess the effect of partial or complete replacement of sucrose with prebiotic polysaccharides, namely inulin or polydextrose, on the rheological characteristics of starch-milk desserts. These polysaccharides were selected because both of them exhibit prebiotic properties, but differ in terms of their chemical moiety as well as structure which may have a significant impact on the rheological characteristics of the desserts.The samples were starch-milk desserts being prepared from potato starch  and from other ingredients, namely powdered defatted agglomerated milk , sucrose , steviol glycosides , curcumin , vanilla flavouring agent , inulin GR , polydextrose .The desserts were prepared according to the recipes given in Pasting properties were determined using a Rapid Visco Analyzer . Milk suspensions of starch with all the other ingredients (as described in apη—apparent viscosity (Pa·s), K—consistency coefficient (Pa·sn), n—flow index (dimensionless).The rheological characterisation of the starch deserts was performed using a Mars II rheometer  with cone-plate measuring geometry . Viscosity curves were determined at 25 °C at increasing shear rate from 1 1/s to 100 1/s, and described by a power law model (1):J(t)—compliance (1/Pa), J0—instantaneous compliance (1/Pa), J1—viscoelastic compliance (1/Pa), retλ—retardation time (s), η0—zero shear viscosity (Pa·s), t—real time of experiment (s), t1—time when the constant stress is removed (s).The creep and recovery tests were performed at 25 °C. The constant shear stress in the creep phase amounted to 1.0 Pa in the range of stress to strain proportionality. The creep and recovery phases lasted for 150 and 300 s, respectively. The resulting curves of compliance as a function of time were described by the Burgers model of four parameters (2 and 3), which consists of the Maxwell and the Kelvin models placed in series:G′) and viscous modulus (G″) as a function of angular frequency (ω). The following power law equations were fitted to the experimental data:G′—storage modulus (Pa), G″—loss modulus (Pa), ω—angular frequency (rad/s), K′, K″—experimental constants, n′, n″—slopes of the plots.A sweep frequency test was performed in the linear viscoelastic range at the constant stain value of 0.03 and in the angular frequency range of 1–100 rad/s. The mechanical spectra were plotted in terms of elastic modulus  followed by the Fisher’s test to establish the least significant difference (LSD) were performed using Statistica 9.0 software . Significance level was set at 0.05 and statistical differences were considered as significant for Viscoelastic structure of milk-starch dessert is formed as a result of heating while stirring and then cooling the milk suspension of starch and other dessert ingredients. Substantial factors determining the structure of the final product and its characteristics perceived by consumers are mainly the amount and kind of starch, interactions between starch and other food components, and thermal processing conditions . PastingThis indicates that a restriction in swelling of the starch granules occurred in the systems due to the higher starch concentration  and from the presence of non-starch components, i.e., sucrose and/or inulin or polydextrose, as well as milk. As a result, the tested systems achieved the maximum viscosity not before a temperature of 95 °C, which is much higher than that achieved by potato starch pasting in water . InteracSubstitution of part of sucrose amount with prebiotic polysaccharides had different effect on the starch pasting . The pasThe increase in the HPV value in systems containing polydextrose could be explained by the structure of this polymer, i.e., a significant degree of branching, which even at elevated temperature could disturb the sample flow, resulting in recorded high viscosity. A significant effect of prebiotic polysaccharides was also observed in the case of viscosity breakdown (BD) during heating . The preK), which reflects the initial viscosity of the material. Additionally, a simultaneous significant decrease in the value of the flow behaviour index (n) indicating an increase in a pseudoplasticity of the dessert was determined. In turn, the complete replacement of sucrose and the introduction of inulin instead resulted in a substantial decrease in the K value and a significant increase in the n value . Substitution of sucrose with polydextrose at all levels resulted in a decrease in the value of viscoelastic compliance as compared to the control sample, and a significant decrease in the value of instantaneous compliance was found only in the case of the highest ratio of polydextrose in the dessert (PD100). Presence of polydextrose in the systems had little effect on the value of zero shear viscosity and only for the PD67 sample a significantly higher value as compared to the control one was found. The starch-milk desserts with polydextrose were characterized by insignificantly lower values of retardation times as compared to the control sample. The results of two-way analysis of variance showed that both the level and the type of sucrose substituting agent had a significant impact on the compliance values . On the G′) and loss (G″) moduli on the angular frequency (ω) for the control dessert and these with inulin (IN) or polydextrose (PD). In all cases, the values of the storage modulus were higher than those of the loss modulus (G′ > G″), indicating the predominance of elastic properties over the viscous ones, wherein a significant dependence of the both moduli on the angular frequency was found. These observations are confirmed by the values of the tangent of phase shift angle (tan δ), which ranged from 0.1 to 1.0, as shown in G′ value with the proportion of inulin in the system. However, the influence of inulin on the G″ value was small. This is confirmed by the value of the tangent of phase shift angle, which decreased significantly with the amount of inulin in the dessert . This increase was much lower than in the case of the use of inulin. The values of the n′ parameter of the desserts containing polydextrose decreased with increasing concentration of the latter, indicating a lower dependence of the G′ modulus on the angular frequency. However, the decrease was much lower than that of the desserts containing inulin. In the case of the K″ parameter, which reflects the initial value of the loss modulus, its slight increase was found compared to the control sample. The statistical differences in the values of this parameter within the samples were, in most cases, insignificant.This means that the analyzed desserts were characterized by typical weak gel properties, which are characteristic for many food systems. Substitution of sucrose with inulin resulted in a significant increase in the  dessert . It indil curves . Substitn″ parameter were lower compared to the control sample. Nevertheless, the decrease in the value of this parameter did not increase with the concentration of the prebiotic polysaccharide in the dessert. The results of two-way analysis of variance showed that both the level and the type of sucrose substituting agent had a significant impact on the values of parameters of the power law models describing the mechanical spectra, with exception of the K″ parameter (J0) and the K′ (r = –0.97) and K″ (r = –0.69) parameters, while the n′ and n″ parameters positively correlated with the instantaneous compliance . Similarly, the values of viscoelastic compliance (J1) negatively correlated with the K′ and K″ parameters , while positively correlated with the n′ (r = 0.95) and n″ (r = 0.94) parameters. It was also found, that the zero-shear viscosity (η0) positively correlated with the K′ parameter (r = 0.88) and negatively correlated with the n′ and n″ parameters .On the other hand, the values of the arameter . AlthougG′ and G″ values and a significant reduction in the tan δ value were observed. Long non-branched chains of inulin can interact with other ingredients of desserts, for example with linear amylose, and through the network of hydrogen bonds can stabilize the structure of the product and enhance the elasticity of the system. Another mechanism of interactions will take place in the case of polydextrose, the structure of which consists of randomly connected and branched short-chain polymers. It will co-create viscoelastic structures through hydrogen bonds in a limited way. Thus, the formation of structures responsible for the increase in proportion of elastic properties in the viscoelastic ones will concern less polydextrose than inulin itself. Polydextrose will generate a thickening effect resulting, among others, from a steric hindrance, a phenomenon characteristic for highly branched polysaccharides.One of the key factors influencing the viscoelastic characteristics of starch foods enriched with inulin is the degree of polymerization of the latter, and thus the length of the fructan chains. Witczak et al.  observedG′), and, at the same time, reduced values of both the tangent of phase shift angle (tan δ) and susceptibility to stress in the creep phase (J). The effect of polydextrose on the viscoelastic characteristics of the desserts was found to be substantially lower than that of inulin itself. The outcomes obtained in the study indicate that the both prebiotic polysaccharides (inulin and polydextrose) seem to be suitable substitutes for sucrose in desserts with reduced sugar content. The influence of these compounds on the rheological characteristics of the product is governed by their structures. Inulin, being a linear polymer, can strengthen the elastic structure of a starch-milk dessert by interacting with its other ingredients, mainly through the formation of hydrogen bonds. Polydextrose, on the other hand, will exhibit a thickening effect due to its more branched structure.Substitution of sucrose with prebiotic polysaccharides modified the rheological characteristics of the starch-milk desserts to a degree depending on the type and level of the substituting agent. The introduction of inulin reduced the peak viscosity of starch paste, which could be related to the restriction of swelling of starch granules and to a plasticizing effect. Inulin had, however, no effect on the final viscosity of the desserts, while polydextrose caused its increase as compared to the control dessert. The desserts showed a non-Newtonian, shear-thinning flow behaviour. The use of inulin, in both the highest and the lowest concentrations, significantly changed the consistency coefficient and the flow index values, while such a phenomenon was not observed in the case of the use of polydextrose. Substitution of sucrose with inulin significantly strengthened the viscoelastic structure of the dessert, which was manifested in increased values of the storage modulus (


Behavioral and Brain Functions (BBF) were made available online [BBF was the first behavioral neuroscience journal that was online-only and open-access (OA). While this may seem to be quite out-of-the-ordinary nowadays, 15 years ago this was an important innovation. Many journals were still paper-only and just a short time before it had become clear that non-paper, “electronic” journals were eligible to be included in MEDLINE/PubMed or the Science Citation Index, or could obtain an impact factor. And although more general online-only OA journals had been established a few years earlier, it was not at all clear whether enough authors would be willing to forego paper and submit to the fledgling journal.Fifteen years ago, on April 22, 2005, the first articles published by the new journal e online . The jouHowever, Terje’s instinct that the time was ripe for an OA journal in the field of behavioral neuroscience proved to be correct and, from the start, the journal received many diverse first rate submissions. By now, 15 years later, the journal has established itself as a well-respected voice in the field, publishing a variety of high-quality articles that are regularly cited in other journals, as shown by its solid current impact factor of 2.125.From the beginning, the journal published research on a diversity of organisms and this has continued to the present day. Indeed, recent published articles have covered research on fruit flies , 4, zebrOver the years, the field has experienced significant growth, as the notion that the most important aspect of brain science is how the brain’s output gets translated into behavior gained more and more traction. To paraphrase Theodosius Dobzhansky: Nothing in neuroscience makes sense except in the light of behavior. As a result, more and more neuroscience studies nowadays include at least some behavior.Behavioral and Brain Functions covers the whole field of behavioral neuroscience, we are especially interested in covering work that addresses some of the key issues that the field is facing. Two of the most pressing issues are the validation of behavioral concepts [most tests were eminently replicable).While concepts  and the concepts . Some ofconcepts . (It shoOther failures to replicate, however, lay on a whole different level. From the results of behavioral tests, both in animals and in humans, we infer behavioral concepts such as spatial learning, anxiety, behavioral despair, etc. However, often enough tests that purportedly tap into the same behavioral construct do not correlate with each other e.g., , indicatFinally, at the occasion of our 15th anniversary, I would like to express my sincere gratitude to all those who have worked tirelessly to get the journal where it currently is: my predecessors as editors-in-chief, those who have served as associate editors, and the members of the editorial board. Many thanks are also due to our authors for submitting their work to us for consideration for publication, but above all I want to recognize our unheralded reviewers, who selflessly donate their time, effort, and expertise to maintain the quality of our journal.While celebrating the successful past 15 years, we look forward with anticipation to the future and further growth of our journal!


PAL1 and PAL2. Regarding the “SG/Adara” graft-combination, there were neither external symptoms of “translocated” incompatibility nor significant differences in the biochemical and molecular parameters between scion and rootstock, proving it to be a compatible combination. The differential expression of PAL genes together with the biochemical factors cited above could be good markers for the “translocated” peach/plum graft-incompatibility.Grafting is an ancient plant propagation technique widely used in horticultural crops, particularly in fruit trees. However, the involvement of two different species in grafting may lead to lack of affinity and severe disorders between the graft components, known as graft-incompatibility. This complex agronomic trait is traditionally classified into two categories: “localized”  and “translocated” . Over the decades, more attention has been given to the different mechanisms underlying the “localized” type of graft-incompatibility; whereas the phenylpropanoid-derived compounds and the differential gene expression associated with the “translocated” graft-incompatibility remain unstudied. Therefore, the aim of this study was to shed light on the biochemical and molecular mechanisms involved in the typical “translocated” graft-incompatibility of peach/plum graft-combinations. In this study, the “Summergrand” (SG) nectarine cultivar was budded on two plum rootstocks: “Adara” and “Damas GF 1869”. “Translocated” symptoms of incompatibility were shown and biochemically characterized in the case of “SG/Damas GF 1869” graft-combination, 3 years after grafting. Non-structural carbohydrates (soluble sugars and starch), phenolic compounds and antioxidant activity, were significantly enhanced in the incompatible graft-combination scion. Similarly, the enzymatic activities of the antioxidant enzyme peroxidase, the phenylalanine ammonia-lyase (PAL) and polyphenol oxidase involved in the phenylpropanoid pathway were significantly affected by the incompatible rootstock “Damas GF 1869”, inducing higher activities in the scion than those induced by the compatible rootstock “Adara”. In addition, a positive and strong correlation was obtained between total phenol content, antioxidant capacity and the expression of the key genes involved in the phenylpropanoid pathway,  Grafting is the association of two genetically different parts: a scion and a rootstock, living in symbiosis to produce a plant with desirable characteristics of both genotypes . The surApart from the total incompatibility characterized by a complete failure of grafting, In spite of the increasing interest of using new rootstocks in most horticultural crops , biochemPAL) genes implies distinct roles in development of graft incompatibility symptoms in different species and could be good markers for the study of compatibility due to its importance in the biosynthesis of many phenolic compounds  in breeding programs for different stone fruit species in Mediterranean temperate areas  in response to compatible and incompatible graft-combinations were performed.Although phenolic compounds and genes related to the “localized” graft-incompatibility have already been studied in different  species , to the P. persica (L.) Batsch] budded on two plum rootstocks: “Adara” (“SG/Adara”) and “Damas GF 1869” (“SG/Damas GF 1869”) grown in nursery conditions. “Adara” was selected as a polyvalent rootstock for different stone fruit species . “Damas GF 1869” is a pentaploid hybrid plum rootstock (P. domestica × P. spinosa) showing graft-incompatibility with most nectarine and a few peach varieties  for each graft-combination.The present study was performed in the “Summergrand” nectarine cultivar [ species  from an arieties . The twon = 3) at ± 5 cm approximately above (scion) and below (rootstock) the graft-union for each graft-combination. The samples were frozen immediately in liquid N2, lyophilized, ground, and stored at −20°C until further analysis.Bark tissue samples were collected at leaf fall period (October 2017) and during the most active and strong vegetative period (June 2018) from the stem of each individual tree  and kept overnight. The mixture was centrifuged at 20,000 g for 30 min at 4°C . The supernatant was collected and kept as a stock solution at −20°C. Two hundred microliters of each stock solution were vacuum concentrated in a SpeedVac (Thermo Savant SPD111V) and dry extracts were re-suspended to 200 μL of Milli-Q water. Then, concentrations of the main soluble sugars were determined by high-performance liquid chromatography HPLC  with a refractive index detector at 35°C  as previously reported , 2019. F2 in 0.05 N HCl). After 15 min, the starch content was determined spectrophotometrically at 600 nm. The results were expressed as mg starch per g of DW.The starch extraction was performed according to From the previously described methanol stock solution, a water dilution 1:10 was prepared and stored at −20°C until further analysis. The analysis of total phenolic content (TPC), flavonoids, anthocyanins and the relative antioxidant capacity (RAC) were carried out using a spectrophotometer Asys UVM 340 Microplate Reader  as previously reported .Total phenolics content (TPC) was determined using the Folin-Ciocalteau reagent according to the method of Three biological replications of the lyophilized samples (10 mg) were mixed with 10 mL of sodium phosphate buffer , containing 1% of polyvinylpyrrolidone (PVP) and 0.5% of Triton X-100. Phenol absorbents (PVP) and detergents such as Triton X-100 were used in the enzymes extraction to prevent the reaction between enzymes and phenolic compounds during extraction (trans-cinnamic acid liberated per g of dry weight (μmol trans-cinnamic acid/h g DW).The Phenylalanine ammonia-lyase (PAL) activity was determined by the conversion of L-phenylalanine to trans-cinnamic acid according to the protocol of The polyphenol oxidase (PPO) activity was assayed spectrophotometrically based on the oxidation of catechol to 1.2-benzoquinone . The rea3H8O2) in the presence of hydrogen peroxide (H2O2) . The reaction rate of POX was estimated with an initial rate of increase in absorbance at 470 nm. One unit of the enzymatic activity (U) was defined as an absorbance increase of 0.1 per minute under the assay conditions.The peroxidase (POX) activity was determined by the oxidation of guaiacol (Ce (H2O2) . For the® Safe DNA gel stain  and the quantification was determined by an ultraviolet (UV) spectrophotometer . Prior to cDNA synthesis, the endonuclease DNasa I RNasa-free (ref: EN0521) was used to digest strand DNA, according to the manufacturer’s instructions . The total RNA (1 μg) was reverse transcribed using an oligo (dT) 20 and SuperScript® III First Strand cDNA synthesis kit in accordance with the manufacturer’s instructions . Real-Time qPCR amplifications were performed on a 7,500 Fast Real Time PCR System  using specific primers for PAL1 (ppa002099m) and PAL2 (ppa002328m) as described by ® Green PCR master mix (Applied Biosystems), 10 ng of cDNA and 300 nM of primers. Three technical replications for each of the three biological replicates were analyzed. Four candidate reference genes: Actin (ACT11), translation elongation factor (TEF2), RNA polymerase II (RP II), and EST (Gene Bank accession No. DY652828) were tested to identify the most stable reference gene for the normalization of gene expression using a NormFinder algorithm . Correlations between biochemical and molecular parameters, to reveal possible associations, were determined by the average of the three biological replicates, using the Pearson correlation coefficient at P ≤ 0.05. Pearson’s rank correlation matrix (P ≤ 0.05) was done using the R corrplot package. Principal components analysis (PCA) was performed and a biplot PCA was designed with combined data from both sampling periods using the SPSS software.Means from three biological replicates for each graft-combination and ungrafted rootstock were analyzed statistically using IBM SPSS 24.0 (United States) software. Data were evaluated by the analysis of variance (ANOVA). When the F test was significant, the separation of means was performed using Duncan’s multiple range test  . These sSugars profile analysis showed that sorbitol and sucrose were the individual soluble sugars identified with higher concentrations in the bark stem of scion and rootstock for both graft-combinations and periods evaluated ,B. OtherThe phenolic compounds content (TPC and flavonoids) and relative antioxidant capacity (RAC) were significantly affected by the “translocated” peach/plum graft-incompatibility. While no significant differences were shown between scion and rootstock in the compatible graft-combination , the incEnzymatic assays showed that PAL, PPO, and POX activities were significantly affected by the incidence of the “translocated” graft-incompatibility. While no significant differences were shown, in general, between scion and rootstock in the compatible graft-combination, the incompatible rootstock “Damas GF 1869” increased significantly the PPO, POX, and PAL activity in the scion at leaf fall . MoreovePAL isoforms (PAL1 and PAL2) was performed in this study. The NormFinder algorithm, used to identify the stability of the four reference genes expression assessed, indicated that the most stable gene under the experimental conditions was RPII  induced approximately three-fold more expressed the PAL1 gene in the scion than the compatible one in both sampling periods  and six-fold (in June) higher expression in the scion when grafted on “Damas GF 1869” than on the compatible rootstock  than at leaf fall period (October). Regarding the compatible graft-combination, no significant differences of PAL genes expression were found between scion and rootstock in both sampling periods. Likewise, there were no significant differences between the ungrafted rootstocks , showing similar values as the compatible graft-combination (“S/Adara”).To identify putative regulation of the PAL activity, a gene expression analysis of two was RPII . Thus, t periods . A more ootstock . For theSignificant correlations were found between the different biochemical and molecular parameters measured at both periods ,3.r = 0.698; P ≤ 0.01), flavonoids , and RAC  at the vegetative period. These correlations suggest that the change in the PAL activity might be a principal factor in the accumulation of phenolic compounds in the scion of the incompatible graft-combination. Moreover, a positive correlation was found between the PPO enzymatic activity and TPC , and RAC  at leaf fall , PPO , and PAL activities , and PAL2 gene expression  at the leaf fall period. Fructose also showed positive correlations with TPC , flavonoids , and RAC  at the vegetative period .Some non-structural carbohydrates were also positively correlated with the phenolic compounds, antioxidant capacity, PAL activity and e period . SimilarPAL genes expression. In turn, the rootstock part (found at the opposite side of PC1 and PC2 at the vegetative period), indicated lower values for carbohydrates, antioxidants compounds and enzymes, showing the reciprocal effect of scion incompatibility. Moreover, whereas for the incompatible graft-combination (“SG/Damas GF 1869”), scion and rootstock were grouped separately, in the case of the compatible graft-combination (“S/Adara”), both scion and rootstock were found in the same group. The ungrafted rootstocks seem to be less explained by the PC1 loadings compared to the PC2 ones.The principal components analysis (PCA) showed that more than 50% of the observed variance could be explained by the first two components in both sampling periods . InteresThe establishment of correlations between the biochemical and molecular parameters measured in the scion and rootstock of the two studied graft combinations may reduce the number of characters for screening genotypes or progenies for graft compatibility studies, as reported by Prunus species, graft incompatibility responses can occur from months to years after grafting (P. cerasifera), alteration of the carbohydrate balance between rootstock and scion, due to the effect of graft-incompatibility but also as consequence of grafting interaction. Changes in sugar and starch accumulation were observed in different species during early stages of graft union formation and development, and as indicator of graft incompatibility , flavonoids and the antioxidant capacity (RAC). The relationship between PAL activity, phenolics accumulation, and RAC was reported in peach/Japanese apricot graft-combinations in response to “localized” incompatibility stress  resulted in an enhanced production of phenolic compounds  as well as the antioxidant activity in the scion of the incompatible graft-combination. Further studies in the identification and transcription of genes involved in other metabolic pathways related to the PPO and POX enzymes could be interesting in the investigation of this type of graft-incompatibility in Prunus. The identification of mRNA and small RNA graft-transmissible signals that may play a pivotal role in grafting physiology could be a step further to this investigation.In the present study, it was possible to reveal the good correlation between the biochemical and molecular parameters, and “translocated” incompatibility across two peach/plum graft-combinations. To date, no previous studies have been reported describing the The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.MM and CF conceived and designed the experiments, contributed with reagents, materials, and analysis tools. RA, CF, RG, and MM performed the experiments and analyzed the data. All authors discussed the results, contributed to the manuscript preparation, and read and approved the final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Purpose: To report the recurrence of a macular corneal stromal dystrophy 50 years after penetrating keratoplasty (PKP).Methods: Observational case reportCasedescription: A 76-year-old male patient presented with visual impairment in the right eye (OD) 50 years after PKP in 1962  in 1968) following explosion injury. His visual acuity had already been impaired before the trauma because of bilateral corneal opacities. The central corneal thickness of the graft measured 584 µm (OD) and 544 µm (OS), whilst the peripheral host thickness (8 mm zone), however, was 1233 µm  and 1131 µm . The original graft diameter measured 6 mm in both eyes and the recipient cornea was cloudy and gray. The endothelial cell count was measured centrally . The visual acuity was 20/100 (OD) and 20/40 (OS). After excimerlaser-assisted repeated PKP , the histological analysis of the former graft revealed deposits of acid mucopolysaccharides (AMP) subepithelially, within the interface, in the donor stroma, and in the endothelium, which proved the peripheral recurrence of a macular corneal stromal dystrophy on the graft.Conclusion: Recurrence of macular corneal stromal dystrophy is seldom, but it may occur many decades after PKP. In this patient, the host’s stroma was twice as thick as that of the graft. This may be caused by the active production of acid mucopolysaccharides in the host endothelium with secondary endothelial decompensation. Thus, PKP remains the gold standard in the cure of macular corneal dystrophy for long-term visual rehabilitation. Macular corneal dystrophy is an autosomal-recessive inherited disease affecting the corneal stroma . The dep2 (OD) and 1320 c/mm2 (OS) with a best corrected visual acuity of 20/100 (OD) and 20/40 (OS). The patient was offered a repeated PKP on the worse right eye because of the progressive visual impairment and his discomfort caused by the blurred vision. The size of the new corneal graft was decided to be 8.0 mm, 2.0 mm larger than the former graft 50 years earlier. The cloudy appearance of the patient’s recipient cornea led to the suspicion of a macular corneal dystrophy. Surgery was performed without any complications  and 44 years after PKP in the left eye  following explosion injury. He reported that his visual acuity had already been impaired because of bilateral corneal opacities before the trauma in 1960. There were no corneal diseases in the family anamnesis. According to the patient, annual follow-up examinations were performed by his ophthalmologist during the 50 years after the first PKP. In addition, further controls were carried out in recent years due to primary open angle glaucoma. Cataract surgery was performed externally 42 years after PKP in the right eye. Six years after cataract surgery – in the meantime, a posterior capsulotomy had also been performed – the patient noticed a deterioration of his visual acuity in the right eye. In addition to the follow-up examinations of his ophthalmologist, the patient attended regular follow-up examinations for glaucoma in our department, where deposits in the periphery of the graft were described 48 years after the first PKP upon slit lamp examination. The graft diameter of the former grafts was 6 mm by slit lamp measurement. The central corneal thickness of the graft measured 584 µm (OD) and 544 µm (OS), whilst the peripheral host thickness (8 mm zone), however, was 1233 µm  and 1131 µm . The recipient cornea was cloudy and gray  gene . The disAlthough appearing only after a long time, recurrence of macular corneal dystrophy is inevitable even decades after PKP due to the recurrent deposition of acid mucopolysaccharides within the corneal graft by keratocyte migration. PKP with a normal graft size of 8.0 mm seems to remain the gold standard in the cure of macular corneal dystrophy for long-term visual rehabilitation.The authors declare that they have no competing interests.