As a retired pathologist, I would like to start this text with a pathology-related story about Japan from more than 100 years ago. I read it as a novice pathology trainee and it remained in my memory for many, many years. I hope that you will understand why I decided to retell it here.Becharrlichkeit führt zum Ziel!” .At the end of the nineteenth and the beginning of the twentieth century, Japan opened up to the West, trying to modernize and catch up with the rest of the world. These efforts also included establishing professional contacts with European medicine, and especially with German physicians. One of the Germans invited to visit Japan was Ludwig Aschoff (1866-1942), Chairman of the Institute of Pathology in Freiburg im Breisgau. He accepted the invitation and traveled for a few weeks by boat to Japan. During his visit, he delivered a few lectures, inspected the medical facilities, and made recommendations on how to improve the pathology services. At the end of the visit, he wrote into the visitors’ book the following words in German: “Becharrlichkeit hat zum Ziel geführt!” . In this context, it is worth a mention that Japanese have since then caught up with the Western World, indeed, and received 28 Nobel Prizes, five of which were in physiology and medicine.Several years later, Aschoff was reinvited to see how much Japanese pathology improved. He was most favorably impressed. Under his original exhortation, he added another sentence: “Croatian Medical Journal (CMJ). Obviously, all this would not have been possible without the wide support of Croatian physicians and scientists, and all the others who have donated their free time to publishing the CMJ over the last 30 years. Special credit goes to the Founding Editors, who made the CMJ happen by conceiving the idea of a high-quality medical journal that coincided with the creation of the new state of Croatia; by realizing the need for such a journal in our country; and by translating their vision into printed pages. Without their vision, dedication, and persistence, the CMJ would not have become what it is today.The message of this story about persistence may be applied to the editors, members of editorial board, and the Deans of the four Croatian Medical schools who became the founding fathers of the CMJ for only a very short period of time. Still, my short stint in that position helped maintain the continuity in the life of the Journal and facilitated the transition of the Editorship from one generation to another. Continuity is very important in medical journal publishing, and thus, I am glad that I was invited to give my part to this end. In that short period of time I learned a lot. I was also given the opportunity to see first-hand some of the problems that previous editors experienced and think of some that the future ones might anticipate. I also saw how well organized the entire editorial operation was. I felt reassured that editorial team would function even without my input and continue to do so flawlessly for many years to come. The fact that we are here for 30 years since the day the journal was launched shows that I was correct in my assessment.In comparison with many others, my own contribution to this joint effort was miniscule, as I served as the Co-editor in Chief of the I like quotes, and thus allow me to finish with two quotes that guided me through my life. The first is a quote by the British statesman and writer Benjamin Disraeli. It is a variation on the theme mentioned in the anecdote about Japan and it goes as follows: “The secret of success is consistency of purpose.” If you have a purpose and transmit the belief in your purpose to others the success will follow.Le problème n’est pas comment reussir mais comment durer” (in English translation: “The problem is not how to succeed but how to last”). Since the CMJ has survived already 30 years, one can rightfully predict that it will last for much, much longer. May it live, grow, and flourish, or as one would say it in Latin: Vivat, crescat, floreat!The second quote is from the French writer André Gide: ”


Biocomposites were prepared from poly(lactic acid) and two natural reinforcements, a native starch and sugarcane bagasse fibers. The strength of interfacial adhesion was estimated by model calculations, and local deformation processes were followed by acoustic emission testing. The results showed that the two additives influence properties differently. The strength of interfacial adhesion and thus the extent of reinforcement are similar because of similarities in chemical structure, the large number of OH groups in both reinforcements. Relatively strong interfacial adhesion develops between the components, which renders coupling inefficient. Dissimilar particle characteristics influence local deformation processes considerably. The smaller particle size of starch results in larger debonding stress and thus larger composite strength. The fracture of the bagasse fibers leads to larger energy consumption and to increased impact resistance. Although the environmental benefit of the prepared biocomposites is similar, the overall performance of the bagasse fiber reinforced PLA composites is better than that offered by the PLA/starch composites. The interest in bioplastics has increased enormously in recent years ,2. They Presently, the biopolymer produced and used in the largest quantity is poly(lactic acid) (PLA). Its stiffness and strength compete even with those of engineering plastics, but its physical ageing is fast because of its low glass transition temperature  and its Starch is also a biopolymer produced by plants for energy storage and it is available in large quantities . BecauseIn accordance with the considerations presented above, the goal of this work was to prepare PLA/native starch composites and compare their properties to those modified with a natural reinforcement, sugarcane bagasse fiber. All components are derived from natural resources thus they are biodegradable with advantageous environmental impact. The similarity of their chemical structure also justifies comparison. The majority of publications on PLA/natural fiber composites claim weak interactions between the components, thus an attempt was made to modify them by the addition of a functionalized, maleated PLA polymer. Since both the natural fibers and starch are expected to increase stiffness and strength, the attention is focused mainly on the mechanical properties of the composites, i.e., stiffness, strength, and impact resistance, and efforts were made to explore the mechanism of the deformation and failure of the composites. Some comments on the relevance of the results for practice are also mentioned in the final section of the paper.The following sections summarize the results of the work and include some discussion of the results as well. The chemical and physical structures of the two modifying components are compared in the first section and then their effect on composite properties is shown in the next. The quantitative analysis of the extent of reinforcement is presented in the subsequent section, followed by the discussion of local deformation processes, as well as the consequence of the results for the possible practical application of the composites.The chemical and physical structure of the reinforcement used strongly influence its effect on the matrix polymer and on the properties of the resulting composite . The cheThe phase structure of the two reinforcements also shows some similarities, both are semicrystalline materials containing an ordered, crystalline phase. Both the strong self-interactions and crystallinity result in large stiffness. However, while starch is a relatively pure material, cellulose fibers contain other components as well, mainly hemicellulose, lignin, and waxes ,51. The Particle morphology is crucial for such materials used as reinforcements in polymer composites. Size, shape, and the extent of anisotropy, i.e., aspect ratio, are extremely important in the determination of deformation processes and final properties ,54. The Since fiber-reinforced materials are mostly used in structural applications, the attention is focused mainly on their mechanical properties. The stiffness of the composites containing various amounts of the two reinforcements is shown in We may assume that the inherent stiffness of starch is smaller than that of the sugarcane bagasse fibers, but the anisotropic particle geometry and the orientation of the fibers must also contribute to the larger stiffness of their composites. Using MAPLA as coupling agent in the composites does not improve or deteriorate stiffness that is not very surprising, since stiffness is not very sensitive to changes in interfacial adhesion ,57.The deformability of the composites (not shown) decreases marginally with an increasing amount of the additives independently of their type or coupling. The strength of the composites containing the two reinforcements is presented in Another surprising result is that the MAPLA coupling agent practically does not affect the tensile strength of the composites either, although properties measured at larger deformations depend on interfacial adhesion quite strongly . In spitImpact resistance is another mechanical property that is important for most structural materials; generally large strength and impact resistance is required from such materials used in practice ,60. The Tσ shows the true tensile strength of the composite; Tσ0 is the same for the matrix, fφ shows the volume fraction of the second component, and B is the load-bearing capacity of the reinforcement. The latter depends on interfacial adhesion. True tensile strength  expresses the change in the cross-section of the specimen during deformation and nλ takes into account strain hardening (n can be determined from matrix properties and characterizes the strain hardening tendency. Reduced tensile strength can be expressed by the rearrangement of Equation (1)The reinforcing effect of additives is difficult to assess based on the primary results of stiffness and especially strength values even though it is almost invariably done so. Reinforcement depends on several factors including the spatial arrangements of the fibers, their size, shape, and orientation as well as on interfacial interactions. Mathematical models give a quantitative estimation of the reinforcing effect of the most various second components. Such a model was developed earlier, which describes the composition dependence of tensile strength . The modThe natural logarithm of tensile strength can be plotted against composition and the plot should result a straight line. The slope of the line expresses the reinforcing effect of the additives quantitatively. The tensile strength of the composites is plotted in the way indicated by Equation (2) in B and the orientation of bagasse fibers, which compensates for the negative effect of large size. The analysis indicates that the load-bearing capacity of the two reinforcements is similar in spite of the different primary values shown in 2 was measured for starch, while 38 mJ/m2 for bagasse fibers by inverse gas chromatography leading to similar work of adhesions (99.4 mJ/m2 and 100.2 mJ/m2 respectively), i.e., strength of interfacial adhesion.The strength of interfacial adhesion is difficult to predict from the model because of the different size and shape of the particles, but it can be estimated by the reversible work of adhesion. This latter can be calculated from the surface tension of the components. The matrix of the composites was the same, and the surface tension of the two reinforcements is quite similar, 35 mJ/mThe results presented above indicate that considerable differences exist in the properties of the composites prepared with the two kinds of reinforcements. Stiffness and impact resistance was larger for the PLA/bagasse fiber composites that is a slight contradiction in itself since larger stiffness is usually accompanied by smaller impact strength , while tComposites are heterogeneous materials containing components with dissimilar elastic properties. External load results in the development of local stress maxima, which initiate local deformation processes. These processes like the separation of the interface between the matrix and the reinforcement (debonding), and the fracture of the particles can be followed by various techniques including volume strain measurements, or by acoustic emission tests. The results of the latter measurements are presented in The comparison of the two plots presented in AE1 and σAE2, are plotted against composition in The plotting of the cumulative number of signal traces allows the determination of characteristic stresses which can be assigned to the local processes taking place during the deformation of the specimen. The determination of these values from the plots is shown in Fracture is difficult to imagine for starch particles, but it is not impossible. The stress belonging to fiber fracture is much larger and can be clearly separated from the first process for the bagasse composites. The existence of a second, competitive process cannot be identified definitely for starch-filled composites but SEM micrographs may provide further evidence about the processes occurring during failure. SEM micrographs recorded on the fracture surface of specimens broken during tensile tests are presented in AE1) and thus larger composite strength compared to composites containing bagasse fibers. The presumably larger inherent strength of the fibers leads to considerably larger impact resistance for the PLA/bagasse fiber composites. Because of the similar interfacial interactions of the two reinforcements, interfacial adhesion is also very similar for the two  was provided by NatureWorks LLC . The extrusion grade polymer (<2% D isomer) has a density of 1.24 g/cm3, and its MFR is 3.9 g/10 min (190 °C and 2.16 kg). The production method of the maleic anhydride grafted PLA (MAPLA) coupling agent was described earlier in detail [The matrix, poly(lactic acid)  before extrusion  to eliminate their residual moisture content. PLA and MAPLA were dried together in a vacuum oven at 100 °C and 150 mbar pressure. A Brabender DSK 42/7  twin-screw compounder was employed for the appropriate homogenization of the components at the set temperatures of 170–180–185–190 °C and the rate of homogenization of 40 rpm. The granules were injection molded into tensile bars according to the ISO 527 1A standard using a Demag IntElect 50/330-100 electric injection molding machine. The temperature profile of the barrel was the same as in extrusion, while injection pressure (800–1200 bar) depended on the amount of the reinforcement added. The value of the most important processing parameters were: injection speed: 50 mm/s; holding pressure: 650–800 bar; holding time: 15 s; cooling time: 45 s. The temperature of the mold was set to 20 °C. The specimens were kept under ambient conditions  until further testing.An Instron 5566 universal testing machine  was used for the characterization of tensile properties with a gauge length of 115 mm and 5 mm/min crosshead speed. An acoustic emission (AE) equipment was used simultaneously with tensile testing to detect local deformation processes. The specific equipment used was a Sensophone AED 404 apparatus manufactured by Geréb és Társa Ltd. . A single piezoelectric sensor (150 kHz resonance frequency) was attached to the center of the specimen. The threshold level of detection was set to 25 dB. A Ceast Resil 5.5 impact tester  was used for the determination of Charpy impact strength (ISO 179 standard at 23 °C with 2 mm notch depth) of the specimens. Fracture surfaces were studied by scanning electron microscopy . Micrographs were recorded on surfaces created during tensile and fracture testing, respectively. Accelerating voltage (5 and 15 kV) and distance to the sample changed according to magnification and the quality of the image. Detectors were used both for secondary and back-scattered electrons. Surfaces were sputtered with gold before the recording of micrographs from the composites. The particle size and size distribution of the modifying components were determined by laser light scattering using a Horiba LA 950 A2  analyzer. The size, size distribution, and aspect ratio of the bagasse fibers were also determined after processing by digital optical microscopy . The polymer was dissolved in tetrahydrofuran  to extract the fibers from the composites.The comparison of the properties of PLA composites reinforced with native starch and sugarcane bagasse fibers shows that the two additives influence properties differently. The analysis of their chemical and physical structure indicates that the former is quite similar, while considerable differences exist in the latter. Particle characteristics differ even more and are shown to influence properties considerably. The strength of interfacial adhesion, and thus the extent of reinforcement, are similar because of the similarities in the chemical structure of the reinforcements. Relatively strong interfacial adhesion develops between the components which renders coupling inefficient. Dissimilar particle characteristics, on the other hand, influence local deformation processes considerably. The smaller particle size of starch results in larger debonding stress and thus larger composite strength. Besides debonding, considerable fiber or particle fracture also takes place during the failure of the composites. The larger inherent strength of the bagasse fibers leads to larger energy consumption during fracture and increased impact resistance. Although the environmental benefit of the prepared biocomposites is very similar, the overall performance of the bagasse fibers reinforced PLA composites is better than that offered by the PLA/starch composites.


Vibrio alginolyticus LHF01 was engineered to efficiently produce poly-3-hydroxybutyrate (PHB) from starch in this study. Firstly, the ability of Vibrio alginolyticus LHF01 to directly accumulate PHB using soluble starch as the carbon source was explored, and the highest PHB titer of 2.06 g/L was obtained in 18 h shake flask cultivation. Then, with the analysis of genomic information of V. alginolyticus LHF01, the PHB synthesis operon and amylase genes were identified. Subsequently, the effects of overexpressing PHB synthesis operon and amylase on PHB production were studied. Especially, with the co-expression of PHB synthesis operon and amylase, the starch consumption rate was improved and the PHB titer was more than doubled. The addition of 20 g/L insoluble corn starch could be exhausted in 6-7 h cultivation, and the PHB titer was 4.32 g/L. To the best of our knowledge, V. alginolyticus was firstly engineered to produce PHB with the direct utilization of starch, and this stain can be considered as a novel host to produce PHB using starch as the raw material. Escherichia coli has been extensively studied as the host bacterium, but the range and efficiency of the biosynthesis applications begin to be limited due to its intrinsic capabilities. Interestingly, Vibrio strains are gaining more and more attentions due to their rapid growth rates. For example, the doubling time of Vibrio alginolyticus and Vibrio parahaemolyticus is reported to be 12-14 min  is an environment-friendly alternative to replace petroleum-based plastics due to its biodegradability, and the most common of which is poly-3-hydroxybutyrate (PHB) . Generalcultures . Recentloduction . Studiesoduction . The mosoduction . There aoduction .Geobacillus sp.  medium, while V. alginolyticus was cultured in TYS50 medium. TYS50 medium contained (g/L) NaCl 50, KCl 0.7, CaCl2·2H2O 1.4, MgSO4·7H2O 6.8, MgCl2·6H2O 5.4, NaHCO3 0.2, yeast extract 1, and peptone 5. When required, 1.5% (w/v) agar was added to obtain the corresponding solid medium. In the process of culturing V. alginolyticus LHF01 for PHB production, soluble starch was firstly used as carbon source, and the shake flask experiments were performed at 37°C and 200 rpm. The culture conditions were optimized in terms of pH, soluble starch concentration, and culture time. The pH was set at 5, 6, 6.5, 7, 7.5, 8, 8.5, 9, and 10, while the starch concentration gradient was 20, 30, and 40 g/L. The samples were taken every 2 hours to determine the cell growth, PHB accumulation, and starch consumption.During all experiments, V. alginolyticus LHF01 was extracted using a Blood & Cell Culture DNA Midi Kit  according to the manufacturer’s protocol. DNA concentration and purity were determined via Qubit fluorometer and Nanodrop 2000 spectrophotometer . DNA integrity was assessed by 1% agarose gel electrophoresis. Whole genome sequencing was performed on the MGISEQ-2000 platform and PacBio Sequel II system at BGI .The genomic DNA of V. alginolyticus LHF01 by conjugation. In brief, the plasmid was firstly transformed into E. coli S17-1 to prepare the donor strain. E. coli S17-1 and V. alginolyticus strains were cultured in LB and TYS50 medium, respectively, with the relevant antibiotics to an OD600 of 1.5-2. Afterwards, 1.5 mL of donor and recipient cells were respectively placed in a centrifuge tube and centrifuged at 6,000 rpm for 5 min. The supernatant was discarded, and the cell pellets were washed once with 1 mL of the corresponding medium, and then suspended in 50 μL of TYS50 medium at a ratio of 1:1. The suspension was cultured for 4-8 h in a 37°C incubator. Finally, an inoculation loop was dipped into the bacterial solution, and then streaked on the TYS50 medium plate with appropriate antibiotics. The plate was cultured in a 37°C incubator for 36-48 h. A single clone was picked out, and put into the TYS50 medium with antibiotics for 24-48 h, and the bacterial solution was verified by 16S rDNA sequencing and stored for follow-up research.The target plasmid was transferred into After treating with boiling water, the soluble starch can be dissolved and its solution is transparent at room temperature. Since starch is turned to blue when exposed with iodine, its consumption could be indicated by adding the iodine solution. To measure PHB accumulation, the strain culture was centrifuged at 10,000 g for 10 min, and washed twice with deionized water. Then the cell pellet was lyophilized to determine the cell dry weight (CDW) and PHB content. CDW was calculated based on the weight of the empty centrifuge tube, the weight of the centrifuge tube containing the cell pellet, and the volume of freeze-dried culture. The lyophilized cell was placed into an esterification tube, reacted with 2 mL of chloroform and 2 mL of esterification solution at 100°C for 4 h, and then shaken with 1 mL of deionized water, and set for stratification. The chloroform phase was taken for gas chromatography (GC) analysis as reported previously .V. alginolyticus strains when overexpressing the PHB synthesis operon, the wild type LHF01 and engineered recombinants were cultured in TYS50 medium with 20 g/L soluble starch for 18 h and observed by transmission electron microscopy (TEM). The shake flask culture was centrifuged to obtain the cell pellet, washed twice with pre-cooled phosphate buffered solution, slowly added with pre-cooled fixative solution, and then fixed for 12 h to obtain the cell samples. Then, the sample was sliced and observed according to the methods reported previously . From the 4,882 predicted genes identified, a total of 4,639 protein-coding sequences (CDSs) were determined, and 128 tRNA genes, 37 complete rRNA genes , and four ncRNAs were annotated.Whole genome sequencing has the highest resolution in bacterial genetic research, and is widely used in microbial traceability, transmission, and population structure identification. Therefore, the genomic characterization of bler 8.3 . The com plasmid . The fulV. alginolyticus LHF01 was constructed, and results indicated that they are all α-amylase and belong to the glycoside hydrolase family 13  was verified by 16S rDNA sequencing.The genetic modification of host cells is one of the most powerful tools in engineering bacteria for microbial fermentation. To begin with, molecular biology research requires introducing foreign plasmids into the target hosts. Generally, developing DNA transformation methods for non-model organism poses unknown challenges. Fortunately, the broad host pBBR1 origin was reported to be transformable into jugation . No chloolyticus , thus plagenesis . The elet strain . Overexpt strain . For exaendocina . To explphaBAPC operon and the wild type control. As shown in phaBAPC can well promote the conversion of soluble starch to PHB, and it also demonstrates the success of genetic manipulation in V. alginolyticus. In addition, the cells of V. alginolyticus LHF01 and LHF01 (phaBAPC) grown on soluble starch were observed by TEM. It was obvious that electron translucent inclusions can be observed in both bacterial cells, yet LHF01 (phaBAPC) has significantly more intracellular particles than V. alginolyticus LHF01 , LHF01 (03416), and LHF01 (03713) were obtained by conjugation. Shake flask experiments were performed to study the consumption rate of soluble starch. As shown in Next, the three amylase genes of V. alginolyticus LHF01, LHF01 (03713) and LHF01 (03713-phaBAPC) all reached the maximum at 18 h, but it was obvious that LHF01 (03713-phaBAPC) had the PHB highest titer of 4.20 g/L at this time, and the PHB content reached 30.66%  was constructed to explore the synergistic effect of PHB synthesis operon and amylase on substrate consumption and PHB accumulation. The PHB titer of d 30.66% . Furtherus LHF01 . It can V. alginolyticus. Strain LHF01 and LHF01 (03713-phaBAPC) were cultivated in TYS50 medium supplemented with 20 g/L corn starch for 18 h, and starch consumption and PHB titer were recorded. Glucose was also employed as the control. Due to the insolubility of corn starch, the accurate CDW cannot be measured. The PHB titer of wide type strain LHF01 was 1.70 g/L, a litter higher than that of 1.33 g/L when glucose was used as the carbon source. In terms of corn starch consumption, starch was not completely consumed after 8 h cultivation by LHF01, showing that the utilization ability of corn starch was weaker than that of soluble starch. It is worth mentioning that, the PHB titer of LHF01 (03713-phaBAPC) reached 4.32 g/L with corn starch, slightly lower than that of 5.85 g/L using glucose as carbon source (It had to be admitted that the cost of soluble starch is relatively high due to the reliance on chemical or other means to denature starch. Therefore, soluble starch was replaced with insoluble corn starch to explore the direct utilization of starch by n source . In addin source .Aeromonas, Bacillus cereus, Massilia, Halogeometricum borinquense, and Halolamina (E. coli and H. bluephagenesis, which cannot assimilate starch naturally. The recombinant E. coli carrying amylase gene from Panibacillus sp. and PHB synthesis genes from R. eutropha accumulated 1.24 g/L of PHB using starch (H. bluephagenesis harboring an amylase with suitable signal peptide was able to grow on corn starch to 9.5 g/L cell dry weight containing 51.5% PHB (V. alginolyticus LHF01 indicated the fastest starch consumption rate. For example, the engineered V. alginolyticus could exhaust 20 g/L corn starch in 7 h and accumulate 4.32 g/L PHB after 18 h cultivation, while the shake flask experiments for PHA production by other reported strains using starch were usually performed for 48 h or even longer. These results demonstrated V. alginolyticus to be a superior chassis for production of PHB from low-cost corn starch. Further development of genetic manipulation techniques and rational engineering will help to improve PHB production from starch by V. alginolyticus.”Several strains have been reported to utilize starch for PHA production, including lolamina . Generalg starch . Enginee1.5% PHB . CompareV. alginolyticus LHF01, and based on this, the utilization of starch by the strain and the overexpression of PHB synthesis operon and amylase were investigated. V. alginolyticus LHF01 was able to consume 20 g/L soluble starch in 5 h shake flask cultivation, achieving PHB titer of 2.06 g/L. Afterwards, the genome of V. alginolyticus LHF01 was sequenced and analyzed. With the overexpression of its phaBAPC operon, PHB titer was improved to 4.55 g/L, and the consumption of soluble starch was increased with the overexpression of its native amylase. Notably, the engineered V. alginolyticus harboring extra amylase and PHB synthesis operon was successfully used to degrade 20 g/L corn starch in 7 h shake flask cultivation, with PHB titer of 4.32 g/L. The use of V. alginolyticus points to the possibility of rapid cell growth for production of chemicals and biopolymers using starch, a sustainable resource with cost much lower than glucose. Further engineering of V. alginolyticus may open new applications in metabolic engineering and synthetic biology research.In this study, a genetic manipulation platform based on conjugation was established for the first-time in


Juice, as a liquid foodstuff, is subject to spoilage and damage due to complications during transport and storage. The appearance of intact outer packaging often makes spoilage and damage difficult to detect. Therefore, it of particular importance to develop a fast, real-time material to evaluate liquid foodstuffs. In this paper, starch films with pH response characteristics are successfully prepared by inorganic ion modification by utilizing whole starch and amylopectin as raw materials. The mechanical properties, stability properties, hydrophilic properties and pH electrical signal response indices of the films are analyzed and measured. The films exhibit good electrical conductivity values with 1.0 mL of ion addition (10 mmol/L), causing the composite film to respond sensitively to solutions with varying pH values. In the test of spoiled orange juice, the full-component corn starch (CS) film has more sensitive resistance and current responses, which is more conducive for applications in the quality monitoring of juice. The results indicate that modified starch films can potentially be applied in the real-time monitoring of the safety of liquid foodstuffs. Food safety has become one of the most important issues in our daily lives. Food safety threatens human health and causes turbulence in the market economy and the stability of society; thus, food safety has a mutually balanced relationship with society and human beings ,2,3,4,5.Food products are damaged and spoiled during transport and storage due to complications. As a liquid food, fruit juice easily deteriorates because of technical packaging defects. Additionally, the appearance of intact outer packaging often makes damage and spoilage difficult to detect. Thus, the development of rapid, real-time materials for liquid food detection is of great significance. The deterioration of liquid foods essentially affects their pH values; therefore, a range of detection materials and techniques can be developed to pick up this particular signal. Zhang’s research group has reported an intelligent indicator label that can detect the degree of food decay . Due to 3 solution as the additive; this film responds sensitively to the pH value of fruit juice with the fast feedback of electrical signals  images of the four starch films are shown in eviously .3 changed the relative crystallinity and layer spacing of the starch films. 3 increased the relative crystallinity of CS films and increased the layer spacing. The relative crystallinity reached a maximum of 11.52% when the doping amount of AgNO3 was 1.8 mL; additionally, the layer spacing reached a maximum at this time. With the increase in AgNO3, the crystallinity peak of the CS/AgNO3 composite film at 19.6° gradually became obvious. According to 3 solution enhanced the relative crystallinity of CA composite films, and the relative crystallinity increased with the addition increasing amounts. AgNO3 increased the layer spacing of CA films and reached a maximum at 0.5110 nm with an addition amount of 0.6 mL.To understand the crystallinity of the starch, X-ray diffraction (XRD) analysis of the starch was conducted, as shown in + ions was higher than that of the original starch, and the layer spacing increased to varying degrees. The analysis showed that Ag+ ions were enriched around the hydroxyl groups of the starch chains due to polarity, guiding the hydroxyl groups of other chain segments during recrystallization to a certain extent; this phenomenon would increase the proportion of recrystallization and cause an increase in crystallinity after the addition of Ag+ ions. Furthermore, due to the involvement of Ag+ ions in recrystallization, there were other substances in the crystal layer; thus, the layer spacing of the starch crystallization zone increased. The CA film had many crystalline branches within the molecular chains, resulting in weaker intermolecular linkages and lower tensile strength. The CS film was composed of amylose, which presented a helical structure between molecular chains, resulting in interchain bonding and crystallization under the guidance of ions; this film had a higher tensile strength than the CA film. The crystallization characteristics of the two films explained why the mechanical strength of the CS film was higher than that of the CA film in The analysis showed that the recrystallization of starch occurred, and the starch chains changed from disordered to ordered. During starch gelatinization, the ordered starch became disordered under influence of the actions of water and temperature, and the crystalline region opened completely. In starch film preparation, as the temperature decreased, the disordered starch chains in the high-energy state gradually tended to become ordered in the low-energy state due to the action of molecular potential energy, producing the phenomenon of gelation. When the moisture content of the starch film reduced to a certain level, the overall movement of the starch molecular chains substantially reduced or even froze, and the system was in a thermodynamic nonequilibrium state. As a result, the hydrogen bonds within or between the molecular chains rearranged, and the starch recrystallized. During recrystallization, amylose in starch formed a double helical structure; simultaneously, a double helical enriched region formed, which was eventually connected by intermolecular hydrogen bonds. In contrast, the recrystallization of CA occurred between the branches, with more crystalline regions forming within the molecule. Because amylopectin has numerous branched chains, the crystallinity of amylopectin formation exceeded that of the CS containing amylose. Therefore, the crystallinity of the CS film shown in 3, the water contact angle for both starch-based composite films showed a trend of first decreasing and then increasing, reaching the lowest value at the addition of the amount of 0.2 mL. The reason for this phenomenon could be that silver ions were heavily doped at the starch chain segment points, and that the hydrophilic characteristics of ions led to a decrease in the water contact angle of the composite film. When the addition amount exceeded 0.2 mL, the ions aggregated to form a chelated starch gel shell; this aggregation made the starch film more hydrophobic, increasing the water contact angle. The water contact angle of the CS composite films was greater than that of the CA composite films. The higher the content of amylopectin in the film was, the better the hydrophilicity of the composite film and the lower the water contact angle [ct angle .3 composite films showed no significant differences in terms of basic peak shapes from those of pure starch films. The -OH vibration absorption peak was at 3270 cm−1 [−1, and the C=O absorption peak was at 1657 cm−1. The distorted vibrational absorption peak of -OH appeared at 1333 cm−1 [−1, and the vibration absorption peak of the C-C skeleton appeared at 850 cm−1 [3 solution enhanced the -OH peak intensity but did not change the functional group type of the starch film. Compared to the pure starch film, the -OH stretching vibration peak at 3287 cm−1 was blueshifted. The smaller the wavelength number of the absorption peak was, the more intense the hydrogen bonding interactions were in the material; this finding further indicated that the addition of the silver nitrate solution enhanced the hydrogen bonding interactions [Fourier transform infrared (FTIR) analysis was performed to observe the changes in the ion-modified starch groups. As shown in 270 cm−1 , the -CH333 cm−1 . The str850 cm−1 . As obse3 composite film had similar percentages of C and O elements; for amylopectin, the proportion of C in the CA/AgNO3 composite film was greater than that in the pure amylopectin film. As shown in −3 ion had a weak oxidation ability in solution, and the starch film was oxidized and deoxidized during preparation; this phenomenon could reduce the proportion of oxygen [3 composite films only accounted for 4.62% of C1s. The doping of the silver nitrate solution did not change the structure of the functional groups, but the percentage of different bonds in the structure changed significantly.The changes in groups and elements were not well observed by FTIR analysis; therefore, the films were analyzed by X-ray photoelectron spectroscopy (XPS) . The totf oxygen . In part3-1.0 mL composite films. The EIS diagram features a semicircle in the high-frequency region and a straight line in the medium-to-low-frequency regions  was purchased from Guangdong Xilong Chemical Co., Ltd. , AR grade. All solution were prepared with ultrapure water.Corn starch (CS) and corn amylopectin (CA) were purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. , AR grade. Silver nitrate  . The X-ray diffractometer (XRD), model max220 , with Cu Kα radiation (λ = 1.5406 nm) and the relative crystallinity was analyzed using Jade 6 software . The morphology and microstructure of the films were observed by scanning electron microscopy (SEM). The mechanical properties of the film, namely tensile strength and elongation at break, were measured by XLM (PC) with intelligent electronic tensile testing machine. The hydrophilicity and hydrophobicity of the film samples were tested by a JC2000D3R  water contact angle measuring instrument. The impedance of the dried film samples was measured by CHI760E electrochemical workstation . The test frequency was 0.1 Hz~1 MHz, and the amplitude was 0.01 V. Layer-by-layer self-assembled devices were produced for electrochemical testing. The device was prepared by coating graphite paper and the innermost film sample with insulating collodion. The liquid to be measured was dropped on the surface of the film, and the steady current and resistance were determined using chronoamperometry and electrochemical impedance spectroscopy.The component interaction of film samples was studied with a Fourier transform infrared spectrometer (FTIR)  with a resolution of 4 cm3 composite films were prepared by simple solution casting. The starch solution was prepared by dissolving starch (4 g) in 96 mL of ultrapure water with the aid of stirring and adding 1.4 g of glycerol. The solution was transferred to a 90 °C water bath and stirring was continued until the solution was completely gelatinized. Then, the different levels of AgNO3 solution (10 mmol/L) were added to the starch solution and stirring was continued for another 10 min. The homogeneous film solution was poured into the Petri dish (d = 15 cm) and then baked in a constant temperature oven at 45 °C for 6 h. The composite film peeled off from the Petri dish was coded as starch/AgNO3-x, where x indicates the amount of AgNO3 solution added to the starch film. For example, CS/AgNO3-1.0 means that the amount of AgNO3 solution doped in the CS film is 1.0 mL.The starch/AgNO


In this contemporary world, the words data and sustainability play a crucial role in determining the financial budgets of small and medium-sized enterprises (SMEs). Usually, it is stated that the survival of small and medium-sized enterprises (SMEs) is directly proportional to the growth and sustainability factor of the nation. The economic sustainability of a nation is dependent on appropriate functioning of SMEs. Any kind of direct impact on the working of SMEs will have its impact on the whole economy of a nation. There are different factors such as lack of financial capacity, low market demands, restrictions with regard to the capital, and barriers in the supply chain that affect the sustainability of SMEs worldwide. Nevertheless, small, and medium sized enterprises around the world are greatly investing on skills, innovation, and other capital related resources to mark up the demands of the external market. The main objective of this study is to examine the financial budgets of technology-based SMEs from the perspective of sustainability and big data. For this, the study collects data through a questionnaire from 1,800 Small and Medium Sized Enterprises. Based on a detailed and careful examination of the data, only 1,400 of the responses received were considered valid (79.75%). To test the hypothesis stated, the study employs structural equation modeling. This will help the researcher to examine the direct effect of financial budget and technology adaption of SMEs from the perspective of sustainability and big data. Results of the study stated that SMEs sustainability and big data are directly and positively related to the financial budget planning of technology-based SMEs. The study also found that big data plays an important role in the businesses, specifically for their own growth. Recently, businesses across the world are facing extreme sustainability issues and thereby adversely impacting the economy of the nations. Due to the current societal circumstances, most of the businesses are enforced to formulate new ideas to deal with the problems associated with the pressurized societal situation. Especially, Small and Medium sized Enterprises (SMEs) recorded the most pressing difficulties including financial and Non-financial. There are other factors like limited market demand, supply chain disruption, and trade discontinuation that will impact the economy of the nation . The curVery recently,  providedTo achieve the goal of sustainability, it becomes equally important for the SMEs to predominantly survive and contribute to the total economy of the country. Although there exist several ways that hinder the growth of SMEs, it becomes crucial for an SME to survive by itself and contribute to the whole economy of the country . DespitePreliminary studies of SMEs sustainability primarily focused on exploring factors that affected the growth and development of small and medium sized enterprises. Factors such as adopting technology in the SMEs, implementing financial budgets, appropriating financial planning, effective big data management, and so forth affected the growth and functioning of SMEs . A growiPast studies confirmed that data is regarded as one of valued assets for any business, and it's been increasing rapidly day-by-day. This valued asset is popularly termed as big data that comes with 3 peculiar features. These three features are identified as having high volumes of data, variety of data types, and velocity to process the data. Almost all the organizations in the world are striving hard to discover all the possible ways to utilize this big data and develop their business. Majority of the studies confirmed that big data is quite important in the growth and development of a business . It was The growth of small and medium sized enterprises is directly related to the aspect of finance. Innovations with regard to finance, especially in the smooth functioning of SMEs play a critical role in various forms. In order to perform well, external and internal financial resources are necessary within an SME . GeneralRecent studies discussed about the effectiveness of the integration of e-commerce within Small and Medium sized Enterprises and found that latest technologies like mobile banking permits many SMEs the flexibility to settle down their financial transactions through the application . These nH1: There is a positive relationship between financing and SMEs sustainability.Previous studies have shown that technology plays a crucial role in the effective functioning of an enterprise. Adapting to the innovative technology will permit the business entity to establish their brand in the market. It will also boost the competitor factor within the SMEs in the global market . As a reFew other studies stressed that certain SMEs export their products/services at international level. They broaden their market with the help of innovative technologies. Technology based SMEs focused on pushing themselves to enter international market and expand their potential. When the SMEs are exposed to the overseas market, equivalently they et an opportunity to expand themselves and efficiently communicate, transport, and trade their services –22. AbovAnother important aspect of using technology in SMEs is that it gives excellent outcome in the market. Use of advanced technology strengthen the condition of firms in the markets at international and national level . Recent H2: There is a positive relationship between adopting technology and SMEs sustainability.There exists a considerable body of literature on the importance of data within the companies. Data is regarded as an important asset of SMEs. SMEs across the world are exploring different ways to tackle the data, which is usually called as big data. Big data is not only used in large multinational organizations, but also in SMEs. With the aim of improving the quality of a business, SMEs make use of big data to make accurate and quick decision in the need of hour , 28. TheA series of recent studies has indicated that multinational organizations are buying big data. Even startups are purchasing big data technologies to understand the new market, new clients, new business and so forth. Big data will also help SMEs to cut down the costs of business and enter the market. Transforming the big data to business applications will cost them real difficulty. Investing capital in the big data hardware and software is high. Few other studies revealed that technology adaptation, financial innovations, and big data are regarded as the strong pillars of SME , 16. In H3: There is a positive relationship between Big Data and SMEs sustainability.H4: Big Data increases the growth and development of SMEs.H5: Innovative Finance, Technology adaptation and Big Data technologies are essential in SMEs sustainability.The current study mainly concentrates on examining the financial budgets of technology-based SMEs from the perspective of sustainability and big data. It also examines the relationship between innovative finance and SMEs, adapting new technologies within the SMEs, adopting to big data technologies from the perspective of SME sustainability. The analysis of the current study has been conducted through secondary literature and primary analysis as well. The study has developed hypothesis based on the analysis and review of the secondary literature. The formulated hypothesis is tested using a structured questionnaire. Thus, the current study gathers all the required data using questionnaire and hence the study is descriptive. Using the questionnaire's results the researcher will evaluate the hypothesis. This method involves describing the objectives, define the population, select the sample, and later interpret the data and results. Population is a complete set of individuals who are bound together by certain common characteristics. Population is of two types- target and accessible. Target population consists of those whom the researcher wishes to generalize the findings of the study. In the current study the total population include 7.2 million SMEs in China. From the total population, the study derives the required sample. The term sample includes the individuals selected to participate in the study. These selected participants are popularly known as respondents/subjects of research. Sampling methods are of two types–probability/random sampling and Non-probability. To carry out the research, this study employs random sampling method. At times, researchers also tend to use purposive sampling with the aim of obtaining required data from certain area of research.For the purpose of fulfilling the objectives of the study, the study selects the sample consisting of 5,210 SMEs located in China. This study focused on those SMEs in China that were relatively easy to connect using an electronic channel. Prior to the administration of the questionnaire prepared, consent from the SMEs was imitated as the first step. Only after the participants acknowledgment, the questionnaire link was distributed to the selected sample. Out of 5,210 sample, around 4,000 of them acknowledged and agreed to participate in the study. So, in total 4,000 SMEs sent their prior consent, out of which only 2,000 were considered valid. Based on the analysis of the secondary literature, the researcher prepared a structured questionnaire, which addressed the factors like innovations in finance, adapting new technologies within the SMEs, adopting big data within SMEs from the perspective of SME sustainability. This structure questionnaire will allow the selected respondents to share their views, opinions, beliefs, attitudes, and values on the research topic. The data was collected during the period of November 2021 to December 2021 and sample of 3,000 was considered along with their responses. The present identifies different variables like innovative finance, innovative technology, SMEs sustainability, and Big Data technology along with the measurement model. This model is presented in To analyze the data collected, the researcher employs SPSS and AMOS −26.0 version. To study the relationship between the variables identified, the study adopts structural equation modeling. Furthermore, the selected SMEs will be able to provide accurate and precise information on the questions covered in the questionnaires related to the research topic. As the SEM method is highly resilient and efficient in modern times, the researcher decided to use the same. This method will help the researcher to examine the relationship between the variables identified by testing the proposed hypothesis. This method also focuses on the appropriateness of the results and hence was taken into consideration.Although the owners of SMEs were reluctant to share the details of firms such as firm performance, innovations in finance budgeting, adopting innovative technologies, and adopting big data technologies, from the perspective of SME sustainability initially, later, they understood the need to evaluate the financial budgets of technology-based SMEs from the perspective of big data and sustainability and thereby opened, which helped the researcher to record the accurate data. Past researchers also tried to measure the sustainability of SMEs, which were in line with the opinions of the respondents, marked in 5-point likert scale, scaling from extremely poor to extremely good. The study also recorded responses from the participants on innovative ways to manage finance in SMEs from the perspective of sustainability and big data, and even their attitude toward the concept of innovative finance on the 5-point likert scale. Lastly, the study also recorded the responses of the respondents on adapting to innovative technologies and big data within SMEs that will enhance the sustainability on a 5-point likert scale.The current study confirmed the following findings. The validation of the instrument used in the study is recorded in The result of the empirical study shows that innovative ways to manage finance within the SMEs will play a crucial role from the perspective of SME sustainability, as the coefficient value seems to be higher than the cut-off value. Likewise, the results associated with variable adopting technology within the SMEs also confirmed that they seem to significantly impact the smooth functioning of the SMES, especially when advanced technologies are implemented in line with SME sustainability and big data. Al the identified variables seem to efficient and hence the results can be considered as reliable. The findings of the study also disclosed that there is no reliability and validity issue with regard to the identified constructs. The In addition to, the study presented the results in measurement models including all the identified various variables. The study found that the hypothesis proposed in the study have high impact on the SME sustainability. Results of the study document that there is a positive relationship between SME and innovative finance, adapting to technological innovations, adapting to big data technologies, especially from the perspective of SME sustainability and big data. The study also found that the SME in China are high in number and hence the operations of the SMEs should focus on adopting to innovative finance, technology from the perspective of big data and sustainability, which stay significant for the survival of SMEs in this modern world. Even big data technologies will create new challenges and opportunities to the SMEs in contemporary world. Furthermore, there is a positive relationship between adopting technology and SMEs sustainability, which will expose SMEs to new opportunities and challenges. In short, it can be summarized that Innovative Finance, Technology adaptation and Big Data technologies are essential in SMEs sustainability.The present article has proposed and tested the relationship between Innovative Finance, Technology adaptation and Big Data technologies, which play an essential role in SMEs sustainability. Hence this study examined the financial budgets of technology-based SMEs from the perspective of sustainability and big data. The results of the study indicated that there is a positive relationship between SME and innovative finance, adapting to technological innovations, adapting to big data technologies, especially from the perspective of SME sustainability and big data. There is a positive relationship between adopting technology and SMEs sustainability, which will expose SMEs to new opportunities and challenges. Thus, it can be argued that can be summarized that Innovative Finance, Technology adaptation and Big Data technologies are essential in SMEs sustainability.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Prevention of preterm birth:a randomized trial of vaginal compared with intramuscular progesterone. Acta ObstetGynecol Scand 2012; 92:215–222: DOI: wileyonlinelibrary.com), and has been published byagreement between the journal's Editor‐in‐Chief and John Wiley and Sons, Ltd. The notehas been agreed due to concerns raised regarding the authenticity of the data presentedwithin the article. The authors have provided a response to the journal regarding theseallegations. The journal has requested an investigation from the author's institution,University of Menoufia, and an investigation is on‐going. As a result, the journal isissuing this note to readers. A subsequent note will be published depending on theoutcome of the investigation.This note is for the above article, published online on 27 September 2012in Wiley Online Library (


The thickness of neointimal hyperplasia, degree of inflammatory cell infiltration, and degree of collagen deposition were significantly higher at 28 days than at 7 days . The TPU and GL blended NFs-covered stent-grafts successfully maintained the patency for 28 days in the porcine iliac artery. Although thrombosis with neointimal tissue were observed, no subsequent occlusion of the stent-graft was noted until the end of the study. Composite synthetic/natural matrix-covered stent-grafts may be promising for prolonging stent-graft patency.Stent-grafts composed of expanded polytetrafluoroethylene (e-PTFE), polyethylene terephthalate (PET) and polyurethane (PU) are characterized by poor endothelialization, high modulus, and low compliance, leading to thrombosis and intimal hyperplasia. A composite synthetic/natural matrix is considered a promising alternative to conventional synthetic stent-grafts. This study aimed to investigate the efficacy of thermoplastic polyurethane (TPU) and gelatin (GL) blended nanofibers (NFs) covered stent-graft in the porcine iliac artery. Twelve pigs were randomly sacrificed 7 days (n = 6) and 28 days (n = 6) after stent-graft placement. The thrombogenicity score at 28 days was significantly increased compared at 7 days ( Commercially available synthetic stent-grafts composed of expanded polytetrafluoroethylene (e-PTFE), polyethylene terephthalate (PET) and polyurethane (PU) are characterized by poor endothelialization, high modulus, and low compliance, leading to thrombosis and intimal hyperplasia5. Therefore, rapid endothelialization, good mechanical properties, thromboresistivity and biocompatibility are key to preserving the long-term patency of stent-grafts7. A composite synthetic/natural matrix is thought to be a promising alternative to conventional synthetic stent-grafts since it has sufficient mechanical properties and good cytocompatibility8.Synthetic vascular grafts are widely used in surgical and/or minimally invasive interventional therapeutic procedure to treat vascular diseases, such as coarctation of the aorta, aneurysm, arteriovenous fistula, dissection, iatrogenic vascular perforation, traumatic injury, and hemodialysis access9. Several synthetic or natural biomaterials have been used to fabricate NF scaffolds include polylactide, poly-caprolactone, poly (glycolic acid), collagen, chitosan, and gelatin (GL)14. GL is an excellent tissue engineering material owing to its unique features, such as biodegradability, biocompatibility, ability to promote cell adhesion and proliferation, and low immunogenicity16. In addition, previous research has demonstrated the GL’s ability to improve the strength of electrospun matrices17. Meanwhile, Tecoflex is a family of thermoplastic polyurethane (TPU) that would be a good candidate for reinforcement. As a thermoplastic elastomer, TPU has been widely used as covering materials for breast implants, catheters, and prosthetic heart valve leaflets because of its superior mechanical properties18. The matrix composition of the TPU and GL can alter the properties of blended metrics and may enhance the mechanical and biological properties. The optimized TPU and GL blended matrices has high potential as materials for use in engineering of elastic tissues like stent-grafts, valves, patches for vascular and non-vascular organs18. The TPU and GL blended NFs covering membrane have been investigated as composite 3D matrix for stent-grafts; however, this composition is yet to be studied in vivo. Therefore, the purpose of this study was to investigate the efficacy of TPU and GL blended NFs-covered stent-graft in the porcine iliac artery.Nanofibers (NFs) coatings manufactured by electrospinning (ES) can be adequately durable if the covering layer’s appropriate polymeric composition and thickness are designedThe stent  was woven into a tubular configuration with single thread of 0.127-mm-thick nitinol wire. When fully expanded, the stent was 6 mm in diameter and 30 mm in length. Radiopaque markers were attached to the ends and middle of the stent to facilitate visualization under fluoroscopy. The 6-Fr delivery system had a usable length of 500 mm and was comprised of an outer braided sheath and a pusher catheter with a guiding olive tip .16. The stent was secured on the cylindrical Teflon collector   were used in this study and randomly sacrificed 7 days (n = 6) and 28 days (n = 6) after stent-graft placement. The TPU and GL blended NFs-covered stent-graft was placed in the left iliac artery (LIA). All pigs were housed under the same environmental circumstances (temperature of 24°C ± 2 with a 12-h day-night cycle) and were supplied with water and food ad libitum. All animals were euthanized with 75 mg/kg potassium chloride via marginal ear vein injection at 7 or 28 days after stent-graft placement.All pigs were anesthetized intramuscularly with a mixture of 50 mg/kg zolazepam and 50 mg/kg tiletamine  and 10 mg/kg xylazine  before the experiment. An endotracheal tube was then placed, and anesthesia was maintained by inhaling of 0.5–2% isoflurane  with 1:1 oxygen (510 mL/kg/min). A 5-Fr micro puncture kit  was used to puncture the right femoral artery under ultrasonographic guidance . Then, a 7-Fr vascular sheath  was replaced under fluoroscopic guidance . Intra-arterial heparin  was administered directly into the sheath. Pre-procedural angiography was performed to confirm location of the iliac artery, and a 0.018-inch guidewire  was inserted through the sheath and advanced to the femoral artery. The stent delivery system was inserted over the guidewire into the LIA and the stent was placed. Post-procedural angiography was performed to confirm the patency and location of the placed stent-graft. A vascular closure device  was used followed by manual compressed for 10 min. Antibiotics  and analgesia  were routinely administered for 3 days after the procedures.Follow-up angiography via the right carotid artery was performed to evaluate stent patency and stent-related complications at 7 and 28 days after the procedure, respectively. The luminal diameters were measured and compared at the middle portion of the stented iliac arteries using RadiAnt DICOM viewer .19. The samples perfused for 48 h using 4% formalin for fixation. The samples were axially sectioned at the middle portion of the stented LIA. Tissue samples were embedded in paraffin, cut into 5 μm-thick slices and stained with Hematoxylin and Eosin (H&E) and Masson’s trichrome (MT). Histologic evaluation using H&E included determining the thickness of neointimal hyperplasia and degree of inflammatory cell infiltration. The degree of collagen deposition was determined on MT-stained sections. The thickness of neointimal hyperplasia and the degree of inflammatory cell infiltration were calculated as the average of eight values around the circumference. The degree of inflammatory cell infiltration and degree of collagen deposition were subjectively classified into 1, mild; 2, mild to moderate; 3, moderate; 4, moderate to severe; and 5, severe according to the distribution of the inflammatory cells and collagen20. All scan of staining samples was performed using a digital slide scanner . Measurements were obtained with a digital microscope viewer . Analyses of the histological findings were based on the consensus of three observers who were blinded to the experimental groups.Surgical exploration of the abdominal aortic bifurcation and both iliac arteries was performed for gross examination of possible arterial injury and thrombosis following stent placement. Approximately 100 ml of heparin-treated  saline was irrigated into the arteries. And, the stented LIA and normal right iliac arteries were extracted for evaluated histological examinations. The stent-grafts gently removed from the stented LIA for apparent surface thrombus. The degree of thrombosis was determined using a thrombogenicity score into 1 (mild), thrombus nonexistent or minimal; 2 (mild-to-moderate), thrombus minimal, observed to be covering 1–25% of material surface; 3 (moderate), thrombus moderate, observed to be covering 26–50% of material surface; 4 (moderate-to-severe), thrombus severe, observed to be covering 51–75% of material surface; and 5 (severe), thrombus extensive, covers 76–100% of material surfacep value < 0.05 was considered statistically significant. Statistical analyses were performed using the SPSS software .Data were expressed as mean ± standard deviation (SD). Differences between the groups were analyzed using two-sample t-test and Mann–Whitney U test, as appropriate. A This study was approved by the Institutional Animal Care and Use Committee  and conformed to US National Institutes of Health guidelines for humane handling of laboratory animals. The study was carried out in compliance with the ARRIVE guidelines.p < 0.001) and post-procedure  angiographies. The luminal diameter at 7 days follow-up angiography was not significantly different the post-procedure angiography (p > 0.05). Follow-up angiographies showed good patency of the stented LIAs without stent-related complications such as endoleak, migration, collapse and occlusion. However, irregular luminal narrowing caused by thrombosis was observed 7 days  and 28 days  after stent placement.TPU and GL blended NFs-covered stent-grafts were successfully placed in all pigs without procedure-related complications. All pigs survived until the end of the study without further complications. The angiography findings are presented in Fig. p < 0.001) after stent placement     Fig. a. Furthe01) Fig. b and deg01) Fig. c were siThe present study results demonstrated that the patency of the TPU and GL blended NFs-covered stent-graft was well preserved for 28 days; however, thrombus with mild neointimal hyperplasia gradually increased over time. Follow-up angiography revealed irregular luminal narrowing into the stent-graft and the luminal diameter of the stented iliac artery was significantly lower at 28 days than at 7 days of follow-up angiography. However, stent occlusion was not observed in any of the pigs. Consistent with the gross findings, histological results demonstrated significantly increased neointimal hyperplasia with prominent inflammatory cells and collagen deposition in the 28 days group, which correlated with the angiographic findings.21. However, their bulky design can make deliverability to tortuous. Furthermore, stent-in-restenosis was frequently occurred in case of relatively small diameter arteries (< 6 mm) caused by high possibility of calcified or thrombogenicity of the grafts22. The incidence of restenosis rate of stent-graft covered with e-PTFE was 12% with thrombosis within a month23. Wong et al. also reported that the luminal diameter of e-PTFE covered stent-grafts in the porcine carotid arteries was reduced to 67.2% caused by neointimal hyperplasia with inflammatory reaction24. Other complications related with conventional stent-graft include thrombosis, restenosis and delayed endothelialization25. Therefore, developing new synthetic stent-grafts with improved stent patency and hemocompatibility is becoming increasingly important26. Our preliminary study revealed that the TPU and GL blended NFs-covered stent-graft with the optimized mechanical and biological properties was successfully maintained the patency without significant stent-in-restenosis for 28 days. However, long-term inflammatory response with possible complications should be validated for clinical application.Commercially available and conventional synthetic stent-grafts covered by e-PTFE or PET have been commonly used for large diameter arteries (> 6 mm) and applied for treatment of endovascular diseases27. Furthermore, the simplicity of the ES technique for fabricating a tubular scaffold makes it more popular for vascular tissue regeneration applications28. TPU is commonly used in medical tubing because of its high hemocompatibility. In addition, its non-biodegradability, which may also contribute to blood vessel replacement is required29. Radeleff et al. reported the TPU covered stent-grafts compared over self-expandable metallic bare stents in a 28 days porcine model. However, TPU stent-grafts failed to improve patency due to excessive neointimal hyperplasia and subsequent occlusion15. Despite the relatively good cell viability of TPU, its poor adhesion and proliferation of endothelial cells affects the antithrombogenic properties of vascular structures31. In this case, finding an appropriate material to pair with TPU and convert it into a synthetic stent-graft may be a solution to improve its efficacy for endovascular treatment. Recently, in the context of employing at composite mesh as a vascular graft, TPU and GL were mixed to obtain a composite natural/synthetic polymer matrix16. In addition to promoting adhesion and proliferation of endotheliocytes, GL also plays a role in strengthening the electrospun matrices17. Our study successfully fabricated TPU-and GL-blended NFs-covered stent grafts using ES and showed short term patency without stent-related complications such as migration, endoleak and collapse.ES has been widely used to fabricate NF membranes because of it high porosity, large surface area, nanofibrous structure of electrospun scaffolds, and its mimicking of the physical nano features of the native extracellular matrixThis study had several limitations. First, we did not evaluate the biocompatibility and mechanical characterization of TPU and GL blended NF scaffolds in an in vitro. Second, the follow-up duration might be too short to evaluate a meaningful neointima proliferation and thrombosis formation. Further studies with longer follow-up periods are required to confirm the efficacy and safety of TPU and GL blended NFs-covered stent-graft. Third, there was no comparison with commercially available stent-grafts in our study. Fourth, the experiment was conducted in normal iliac arteries. Our findings may not reflect all pathological mechanisms that occur in humans following stent placement. Finally, the number of animals was too small to perform a robust statistical analysis. Although additional studies are required to further validate our current data, our findings provide the basic concept of TPU and GL blended NFs-covered stent-graft in the porcine iliac artery.The TPU and GL blended NFs-covered stent-grafts successfully maintained patency for 28 days without stent graft-related complications in a porcine iliac arterial model. Although thrombosis with neointimal tissue were observed, no subsequent occlusion of the stent-graft was noted until the end of the study. Although further preclinical studies are needed to investigate its efficacy and safety, a composite synthetic/natural matrix-covered stent-graft may be promising for prolonging stent-graft patency.


Brassica napus/Portulaca oleracea (Bn/Po) and Nicotiana benthamiana/Portulaca oleracea (Nb/Po). This special structure mostly appeared in the local area near the grafting union, and the frequency and quantity of the spiraling tracheary element bundles were much higher in the scion than in the rootstock. Nevertheless, only a small portion of Arabidopsis thaliana/Portulaca oleracea (At/Po) interfamilial grafts showed a less spiraled TEs at the grafting union , which is consistent with its growth performance. This study consolidated that spiraling TE bundles were an important indicator for graft incompatibility. The possible reason for the formation of spiraling TE bundles in interfamilial grafts was discussed.In distantly-related plant grafting, incompatibility often occurs between scion and rootstock, resulting in growth stagnation, and eventually graft failure. In this study, we found that an emergent structure, or the spiraling tracheary element (TE) bundles consisting of TE masses occurring at the graft interface, was extensively present in the highly incompatible interfamilial graft of  Grafting is one of the artificial propagation methods of plants in which two different plant segments are mechanically joined together to survive as a new individual after vascular connection and wound healing. This technique was practiced in ancient Greece and China as early as the 5th century BC . At presArabidopsis and tomato, despite the lack of vascular connection, the Arabidopsis scion grafted onto tomato rootstock can still blossom and produce seeds , mechanical mismatch during grafting, pathological infections, growth condition and growth activity of either graft partner could cause the graft failure , Nicotiana benthamiana/Portulaca oleracea (Nb/Po) and Arabidopsis thaliana/Portulaca oleracea (At/Po) combination was described, and the causes of their formation and their influence on the growth of distant grafting were also discussed.To further test whether this particular vascular structure was occurring in other interfamilial grafts, we focused on interfamilial combination involving purslane as rootstock. The existence of spiraling TEs in the interfamilial grafts, Brassica napus), Nicotiana benthamiana, Arabidopsis thaliana and purslane (Portulaca oleracea) were surface-sterilized in chlorine gas for 2 h. Then they were plated on the sterile MS medium containing 3% sucrose (W/V). Sterile petri dishes containing seeds were vertically placed in the growth room for constant temperature growth (22∼23 °C), and the growth condition was set at long-day photoperiod cycle (16 h of light and 8 h of darkness).The seeds of rape . The grafting process was described previously  at 28 °C for 1.5 h. The treated samples were washed in 5% TritonX-100 for 15 min and then in PEM solution for 15 min. Samples were dehydrated step by step (15 min each) with 15%, 30%, 50%, 75% and 100% ethanol. With another two times of absolute ethanol wash, they were dried for 3 h in a −20 °C, using a low-vacuum drier (CHRIST). The dried samples were mounted on the sample holder, and then placed in the ion sputtering apparatus (SC7620 Sputter coater) for conductive gold coating. Examination of the samples was conducted in a VEGA3 TESCAN scanning electron microscope at 20 kv acceleration voltage. In order to count for the number of spiraling bundles, we surveyed the area within a three mm radius centered at the midpoint of the grafting interface. The spiraling bundle was characterized with spiraling tracheary elements plus either a concave hole or convex point, and thus was counted as a spiraling bundle. The Arabidopsis/purslane grafts (n = 67) were surveyed every two days after grafting (DAG), while rape/purslane grafts (n = 35) and Nicotiana/purslane grafts (n = 43) were mainly surveyed after 28 DAG. For each time of collection, graft unions from at least six individual grafts were prepared.Isolation of vasculature was recently described by Bn/Po heterograft  Lk  Lk . Similarbidopsis , at the bidopsis . They webidopsis , in the eligiosa . Usuallynterface , which lnterface . These eArabidopsis or Nicotiana (icotiana , or of qicotiana . The repSpiraling TE bundles existed extensively in incompatible grafts. They restricted the graft growth and potentially blocked the vascular connection, thus could be used an important indicator of incompatibility of distant grafts. In practice, we can use this structure to examine the possible grafting relationship between scion and rootstock. Furthermore, future study could be designed to address the origin and molecular mechanisms of spiraling TE bundles.10.7717/peerj.14020/supp-1Supplemental Information 1Click here for additional data file.


The purpose of this study was to examine the optical properties of translucent zirconia with different thickness during multiple firings. n = 10) were prepared from preshaded and nonshaded blocks. The coloring liquid  was applied with a synthetic nylon brush in the nonshaded group. Then, the specimens were then subjected to 1, 3, and 5 firing times. After consecutive firings, color differences (ΔE) and translucency parameter (TP) were measured. Statistical analysis was carried out by using repeated measure ANOVA followed by Tukey test.  Three different types of translucent zirconia (Vita YZ HT (HT), Vita YZ ST (ST), Vita YZ XT (XT)) with thickness of 0.5, 1, and 1.5 mm were used in the study. 180 disk-shaped specimens (E were significantly affected by the repeated firings. The highest TP was seen in 0.5 mm XT specimen. For all specimens, TP decreases from 1st to 3rd firing cycles, despite TP increases from 3rd to 5th firing cycles. Although there is a significant change in TP values in 0.5 mm thickness, there is no significant change between firing cycles for 1 and 1.5 mm thickness specimens. The highest ΔE value was observed for shaded specimens between 1st and 5th firings. ΔE values were not significantly different between 1 and 3 firings.  ANOVA analysis reveals that TP and ΔE increased as the thicknesses decreased for both types of translucent zirconia specimens tested. Changes in thickness and repeated firings of zirconia specimens affected final color and translucency of zirconia specimens. HT blocks are more affected by consecutive firings than ST and XT blocks. Δ In recent years, patient's demands for esthetic dentistry have increased. To achieve optimal esthetics and meet patients' request, it is essential for restoration to reach natural translucency of the teeth. Improved translucency of blanks and different coloring methods make it possible to produce restoration with natural tooth color and translucency .Translucency of zirconia is mainly dependent on light scattering and thickness . Also, yTranslucent zirconia blanks are introduced to market as nonshaded (white) and preshaded. Preshaded blanks are more advantageous since they have more uniform color than nonshaded ones . ColorinRestorations can be fired 2 to 5 times due to some shape and color corrections to satisfy esthetic demand of both patient and clinician. Besides, multiple firings are generally necessary after occlusal adjustments. Various studies have reported that firing causes color changes due to pigment breakdown during firing , 12. O'BThere is no study regarding repeated firings on optical properties of newly introduced extra- and high-translucent zirconia. Therefore, this study is aim at revealing if multiple firings can alter translucency and color of translucent zirconia. Additionally, this research is aimed at exploring which of the preshaded or shaded provide better optical properties after repeated firings. This research will enable us to see how the zirconia blanks are affected by the firing processes in terms of optical properties thus enabling dental practitioners to have an ideal approach in their clinical and laboratory practice.The initial hypothesis is that repeated firings would not affect the translucency and color of high- and extratranslucent zirconia. The second hypothesis states that there is no significant color and translucency parameter (TP) change between preshaded and shaded blocks.n = 10). These were fabricated from VITA YZ XT (XT), VITA YZ ST (ST), and VITA YZ HT (HT)  with the help of CAD/CAM system . After milling, the coloring liquid  was applied with a synthetic nylon brush in the nonshaded group. Since most of the clinical color selection is from A2 color, this hue was selected. Then, excess water was wiped off and air-dried. Then, the specimens were predried in a drying cabinet for 15 min at 150 C. After drying process, Vita YZ HT (10 hours) and XT (7 hours) were sintered in firing furnace (Vita Zyrcomat 6000 MS) at 1450 C (4 hours 1530 C for Vita YZ ST) as per the manufacturer's recommendations. After sintering, thickness of specimens was checked with digital caliper  with the accuracy of 0.01 mm.The composition, sintering conditions, and manufacturer of the materials used in this study are presented in After the specimens were ultrasonically cleaned in distilled water for 5 min, the translucency and color of the test specimens were evaluated using a spectrophotometer . Every specimen was subjected to repeated firings. Measurements were performed in first, third, and fifth firings by the same operator. Each value was recorded on five different areas of each specimen including the center of specimen .L∗, a∗, b∗) values of each specimen were measured on a black background and on a white background with a spectrophotometer in the wavelength range of visible light, 400 to 700 nm, at 10 nm intervals. D65 illuminant and 2-degree observer were selected.CIE-Lab 2 + (Δa∗)2 + (Δb∗)2]1/2.The color differences between firings were calculated for each specimen on software according to the formula: ΔE) of different numbers of firings at each sample thickness and staining type. In addition, the mean values of different sample thicknesses for each firing number and staining type were compared using one-way ANOVA and the Tukey HSD test. The independent t-test was used to compare the mean values between the two staining types at each sample thickness and the number of individual firings. Cases with a p value below 0.05 were considered as statistically significant results.Statistical analyses were performed with the SPSS version 23.0 . The conformity of the variables to the normal distribution was examined by histogram graphs and Kolmogorov-Smirnov/Shapiro-Wilk test. Mean and standard deviation values were used when presenting descriptive analyzes. ANOVA was performed for repeated measurements, taking into account the number of firings as repeated measurements and block and staining and sample thicknesses as intergroup factors. Considering the significant interaction effects between all factors, in one-sample repeated measurements, ANOVA and Sidak pairwise comparison tests were used to separately compare the mean values of light transmittance and the color change , for 1 and 1.5 mm thickness specimens, there is no significant change between firing cycles (p > 0.05).While the highest TP (27.63 ± 1.10) was seen in 0.5 mm XT specimen, the lowest TP (10.10 ± 0.46) was seen in 1.5 mm HT specimens. In addition, the TP values of the preshaded samples were found to be higher than the shaded samples. It was determined that the TP change was higher in shaded ones with repeated firings, but this change was significant only in 0.5 mm samples and insignificant in other thicknesses (1 and 1.5 mm).E value (7.05 ± 0.19) was observed for shaded specimens between 1st and 5th firings. ΔE values were not significantly different between 1 and 3 firings. ΔE values between 1st and 3rd and 3rd and 5th were below clinically perceivable threshold except between 1st and 5th firings which was above the threshold. ΔE change is higher in extra- and supertranslucent blocks than in translucent blocks.According to E values decrease. It was observed that ΔE change was more in thick samples after the first firing, whereas there was less change in ΔE values in later firings. It was seen that thin samples were more affected by ΔE changes between 3-5 and 3-7 firings.As the thickness increases, ΔIn this study, the initial hypothesis was rejected because different porcelain thickness and number of firing cycles affected the final color and translucency of extra- and high-translucent zirconia. As a result of the repetitive firings, preshaded and shaded exhibited different translucency and color changes; second hypothesis was also rejected.There are several studies reported that repeated firings resulted in changes on optical properties of zirconia –19. On tThe experimental results showed that thickness affected the translucency and color of monolithic zirconia which was consistent with previous studies , 22. In The present study showed that thin specimens are more affected by repeated firings. The repeated firings changed the translucency 0.5 mm thick specimens but had no significant effect on 1 and 1.5 mm thick specimens. TP values are decreased from 1 to 3 firings, while translucency is increased from 3 to 5 firings for all thickness specimens which is consistent with conducted by Moon et al.  and FathUsing preshaded blocks and by staining white monolithic zirconia blocks in coloring liquids are two main approaches for coloring zirconia restorations , 24. ColThis study evaluated that shaded zirconia blocks showed higher color change according to preshaded zirconia. It may be attributed to preshaded zirconia color was homogeneous and more stable than nonshaded zirconia .E values. Thicker specimens were less affected by repetitive firings according to thin specimens. 0.5 mm specimens were prone to color change after repeated firings while there is less color change in specimens with 1 and 1.5 mm thickness. This was consistent with the study of Fathi et al. [E units were determined as a perceptibility threshold [Based on the present study, the thickness of translucent zirconia influenced the Δi et al.  and Farzi et al.  which wahreshold . DentistE between 1 and 5 firings was significantly higher than that between 1 and 3 firings, ΔE between 3 and 5 firings was found least for all types of translucent zirconia blocks. ΔE between 1 and 5 firing cycles was above the clinically acceptable threshold (3.7 ΔE). These results were in accordance with those of Fathi et al. [E between 1 and 3, 1 and 5, and 1 and 7 firings was significantly higher than that between 3 and 5, 3 and 7, and 5 and 7 firings.In the present study, while Δi et al.  who repoPatients' esthetic demand is increasing for more and more translucent and natural restorations. It is essential to select most translucent type of block and appropriate shade and number of firings. It is crucial to minimize the influence of repeated firings which effect the final color and translucency of translucent zirconia. In choosing convenient materials, number of firing cycles and thickness of restoration should be considered.E increased as the thicknesses decreased for all types of translucent zirconia specimens tested. Since HT blocks were more affected by repetitive firings than ST and XT blocks, it can be recommended to be less preferred. Likewise, it should be taken into account that thin and shaded specimens were more affected by the multiple firings. Further studies are suggested to evaluate different types of restorations such as crowns and laminate veneers.Considering the limits of this investigation, the following conclusions were drawn: TP values of preshaded blocks were found higher than shaded blocks. Additionally, it was observed that Δ


In recent years, there has been increasing interest in studying differences in recipient sex in renal disease treatment, access to renal replacement therapy, and subsequent outcomes. Our aim was to find out whether there are differences in outcomes after renal transplantation between female and male kidney transplant recipients in our series, particularly in adults under 60 years of age during long-term follow-up.n = 1,101) to compare graft survival depending on the sex of the recipient in the entire series and patients < 60 years of age (n = 687) during long-term follow-up.This was a retrospective study of our kidney transplant series (p = 0.040). Pre-transplant sensitization  and donor age  were the independent variables associated with graft failure.We observed no association between recipient sex and graft survival throughout the series, regardless of recipient sex. However, adult female recipients under 60 years of age had lower graft survival than male recipients (per se was not related to graft failure, but the greater immunological risk in women and more frequent use of expanded criteria donors in female recipients under 60 years of age were the main factors related to their poorer graft survival. Further studies and new strategies are needed to identify these differences and develop the best approach to address them.Female recipients younger than 60 years of age had lower graft survival than male recipients, although there were no gender differences in graft or patient survival in the overall study population. Recipient sex  There is growing evidence that there are inequalities in various aspects of kidney disease, such as the burden of kidney disease, access to renal replacement therapies (RRT), or their subsequent development, in relation to the gender of the recipient –3. A higSeveral authors and large registries have found no difference in survival between men and women after kidney transplantation, although the effect of the recipient’s sex on graft survival has been disputed in several studies , 4, 6. TWe performed a retrospective analysis on 1,101 adult deceased and living donor kidney transplant recipients at our hospital from January 2000 to January 2019. None of them was a multi-organ transplant. The allocation process during the period of analysis was not based on a computer algorithm but the independent decision of the nephrology staff. Patients gave signed consent for the use of their clinical and personal data for educational and research purposes. Data were taken from our kidney transplant database, which includes variables on donor and recipient demographics, kidney function, infections, cardiovascular disease, development of cancer, and graft failure or death and includes all patients transplanted since the start of our kidney transplant program. All data were extracted from each patient’s medical records and entered into the database by trained personnel. The data were kept confidential in accordance with Spanish law and the Declaration of Helsinki.We analyzed the demographic characteristics of the recipients, such as sex, age, dialysis modality , dialysis time (in months), re-transplantation, previous HLA sensitization, pre-transplant comorbidities such as hypertension (HTN), diabetes mellitus (DM), smoking habits or ischemic cardiomyopathy, body mass index (BMI), and end-stage renal disease (ESRD) etiology. Donor demographics such as sex, age, type of donor brain death donor (DBD), donor after circulatory death , cerebrovascular death, HTN, and serum creatinine were analyzed. Duration of cold ischemia (hours), immunosuppressive treatment (tacrolimus), delayed graft function (DGF), acute rejection (AR) during the first 6 months diagnosed by biopsy, etiology of graft failure, and patient death were also studied.Initially, we considered HLA sensitization in patients with PRA higher than 10% and, after 2012, in patients with anti-HLA higher than 1,500 MFI (Luminex techniques).The immunosuppressive regimen included therapy with a standard dose of a calcineurin inhibitor (tacrolimus or cyclosporine), m-Tor inhibitors (sirolimus or everolimus), mofetyl mycophenolate, or sodium mycophenolate, and steroids in a decreasing dosage. Induction treatment with interleukin-2 receptor antagonist was used in non-extended criteria donors and low immunological risk patients. Thymoglobulin in reduced doses (two to three doses of 1.25 mg/kg every other day) was used for induction in the case of extended criteria donors with a high risk of DGF and a maximum cumulative dose of 6 mg/kg in those patients with high immunological risk .t-test was used to compare the quantitative variables that were normally distributed, and the Mann-Whitney test was used for the non-normally distributed ones. Normality was tested with the Shapiro-Wilk test and the Bartlett test. Graft survival according to recipient sex was calculated using the Kaplan-Meier method, and the null hypothesis was tested using the Log-rank test.Initially, a descriptive analysis of donor and recipient demographics and post-transplant variables was performed. Qualitative variables were described with absolute frequencies and percentages. Quantitative variables were summarized as the mean and standard deviation. A bivariate analysis to compare donor and recipient characteristics as well as the post-transplant variables, depending on the recipient sex, was performed. Associations between qualitative variables were evaluated by Fisher’s exact test. Based on the recipient sex, the Student’s Additionally, Cox regression models were used to evaluate the time to graft failure depending on recipient and donor characteristics. The following were considered as possible explanatory variables in the model: sex, age, BMI, pre-transplant HLA-sensitization, donor sex and age, cardiovascular disease as the cause of donor death, cold ischemia time, and AR episodes in the first 6 months after transplantation. Goodness-of-fit of the models was evaluated by calculating their concordance, that is, the degree to which the models distinguish between patients at higher risk and lower risk . The score test was used to verify the proportional risk assumption . All theP-values less than 0.05 (two-tailed) were considered statistically significant.R software version 4.0.5  was used to perform statistical analysis. The whole study population included 1,101 kidney transplant recipients with a mean follow-up of 7.2 ± 5.9 years (interquartile range: 2 – 11.1), of whom 435 (39.5%) were female recipients and 666 (60.5%) were male recipients.p = 0.014). The main causes of death were cardiovascular diseases and cancer  and infectious diseases, with no statistical differences  was worse in female recipients than in male recipients only during the first 4 years of the follow-up (p < 0.05).The distribution of patients depending on their age at the time of transplantation resulted as follows: 404 patients older than 60 years (37.1%) and 685 patients < 60 years (62.9%). Recipients < 60 years were 277 (40.4%) women and 408 (59.6%) men. In the bivariate analysis, no age differences were found between female and male recipients at the time of transplantation . ESRD dip = 0.037) later in the follow-up period, from the seventh year onward (p = 0.003) (p = 0.027), followed by donor age  (p < 0.001). No differences in graft survival were found between sensitized patients transplanted with grafts from younger donors and non-sensitized recipients transplanted with grafts from donors older than 60 years (p = 0.845). Patient survival was similar in both men and women    . Recipie= 0.017) . We anal= 0.850) , as was In this study, we found that there are differences in graft survival between women and men in patients younger than 60 years, unlike in the older population. This fact led us to consider the need to develop different strategies to improve outcomes in this high-risk population.Numerous differences related to the gender of the recipient have been described in the medical literature. First, the prevalence of CKD is higher in women than in men , a fact We designed this study to investigate whether post-transplant outcomes differ among men and women in our single-center cohort of renal transplant recipients. The ratio of female to male transplanted patients in our series was similar to that described in other studies, with almost 40% of female patients and 60% of male patients, supporting the differential need for RRT according to the gender of the recipient  and rarely extended to other geographical locations or health systems –24, 26. However, our study has allowed us to learn about the state of this issue in our center, where deceased donors are our main source of kidneys, with a high proportion of donors with expanded criteria, and these results could be representative of other transplant programs with similar characteristics and allocation policies to ours. The most important question for us is how to improve graft survival in adult female recipients younger than 60 years. In our opinion, we need protocols that include pre-transplant and post-transplant measures. Before transplantation, sensitized patients should be enrolled in specific programs to achieve the best HLA compatibility, such as the National Priority Allocation System for Hypersensitized Patients based on virtual crossmatching, which has shown excellent results in cadaveric donor transplantation in Spain and the development of similar programs at the regional level . DesensiIn summary, no differences in graft and patient survival were found in the overall study population depending on the sex of the recipient, but the group of female recipients younger than 60 years had lower graft survival at longer follow-up than the male recipients. HLA sensitization and older donors were the main risk factors for poorer graft survival, with both factors being more pronounced in young female recipients. In female recipients younger than 60 years, strategies to improve outcomes are needed to avoid allocation bias and differences in graft survival depending on the gender of the recipient. These differences could be underestimated. Therefore, multicenter and high-quality studies are needed to improve our knowledge of this problem, find the best approach to avoid it, and, finally, improve our outcomes in long-term follow-up.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the study on human participants in accordance with the local legislation and institutional requirements. The patients/participants provided their written informed consent to participate in this study.AS, EG, and JK participated in the research design, the writing of the manuscript, the performance of the research, and in the data analysis. SB, CC, and VE participated in the research design and the writing of the manuscript. JP and PM analyzed the data. BV, MG, and EC made the figures and revised the manuscript. AA drafted and revised the manuscript. All authors approved the final version of the manuscript.