Organ scarcity demands critical decision-making regarding eligible transplant candidates and graft allocation to ensure best benefit from renal transplantation (RTx). Among the controversial relative contraindications is a history of pretransplant malignancy (PTM). While oncological outcomes of PTM-RTx recipients are well described, data on graft-specific outcome are scarce. A retrospective double case control matched pair analysis (60 months follow-up) was carried out and RTx-recipients were stratified for history of PTM. First, PTM-RTx recipients were matched according to age, sex and duration of immunosuppressive therapy. Next, PTM-RTx recipients were matched 1:1 for age, sex and cause of end-stage renal disease. Five-year patient and graft survival as well as oncological outcomes were analyzed. A total of 65 PTM-RTx recipients were identified. Post-RTx recurrence rate was 5%, while 20% developed second de novo malignancy, comparable to 14% in the control group. PTM-RTx recipients had a noticeable lower five-year death-censored as well as overall graft survival and Cox proportional hazard modeling showed a correlation between PTM and inferior graft survival. Although underlying reasons remain not fully understood, this study is the first to show inferior graft survival in PTM-RTx recipients and advocates necessity to focus on more meticulous graft monitoring in PTM recipients in addition to heightened surveillance for cancer recurrence. Renal transplantation (RTx) is currently the only definitive therapy for patients with end-stage renal disease (ESRD), providing the possibility to avoid livelong renal replacement therapy (RRT) . Since RIn an aging society, including RTx-recipients, incidence of cancer increases and several lines of evidence demonstrate an increased risk of cancer and cancer-related death in potential transplant candidates. Currently, 7.0% of all solid organ transplant (SOT) recipients in population-based studies have a PTM . This nuA retrospective single center study with a double case control matched pair analysis and a follow-up period of 60 months after transplantation was conducted. Ethical approval from local ethics committee was obtained . All organs were procured on behalf of Eurotransplant in different hospitals of the Eurotransplant (ET) area. Transplantations were only performed at the Department of General, Visceral and Transplantation Surgery at the University Clinics Münster, Germany. The study was conducted in accordance with the ethical principles of the Declaration of Helsinki. Donor and recipient data were extracted from Eurotransplant Network Information System (ENIS), in-house transplant data files and patient charts . Only deThe initial screening included all RTx recipients at the Department of General, Visceral and Transplant Surgery, University Hospital Münster, Germany between 1 January 2000 and 31 July 2012, with a five-year follow-up ending on 1 July 2017. Exclusion criteria were recipient age < 18 and recipients with a previous history of transplantation of any kind. Remaining patients were stratified based on the history of PTM. For the first analysis, all RTx recipients with a history of PTM were matched 1:1 in a case control matched pair analysis to corresponding recipients without PTM. The data was then compared in terms of incidence of recurrence and de-novo as well as secondary de-novo malignancy  after RTx. Matching criteria were age, sex, and duration of immunosuppressive therapy. Since using time under immunosuppressive therapy as matching criteria allows inclusion of one of the most established risk factors for developing cancer following SOT, it also entails a bias when analyzing patient and graft survival times. Therefore, a second match-pair analysis was conducted. Here, the PTM-RTX cohort was again matched 1:1 to RTx-recipients without PTM to analyze patient and graft survival. Matching criteria in the second matched pair analysis were age, sex and underlying cause of ESRD.The extracted recipients variables included age, sex, body mass index (BMI), cold and warm ischemia time, underlying cause of ESRD , interstitial nephritis, vasculitis, other  and unknown), dialysis vintage, immunosuppressive therapy (duration and protocol), Human Leukocyte Antigen (HLA) mismatch , history of hypertension, diabetes or coronary artery disease (CAD). For the PTM-RTx cohort, the following additional disease specific parameters were retrospectively collected by electronic record review: primary malignancy entity (non-melanoma skin cancers (NMSC), malignant melanoma, urothelial cell, gynecologic, kidney, gastrointestinal, prostate, thyroid, breast, head & neck, lung, hematologic, neuroendocrine tumors (NETs)), tumor nodus metastasis (TNM)-stage, treatment algorithm, time of diagnosis between malignancy and transplantation and oncological follow-up according to current German oncological guidelines. The donor data included age, sex, body mass index  AB0-status, last serum creatinine pre procurement, type of graft donation (donors after brain death (DBD) or living donors (LD)) and possible allocation within the Eurotransplant Senior Program (ESP). Since the Kidney Donor Profile Index (KDPI) and Kidney Donor Risk Index (KDRI) have been recently validated for an ET cohort, both were used to assess likelihood of graft failure, metric for donor characterization and risk .RTx-recipients received grafts from DBD or LD. Grafts were allocated AB0- and HLA compatibly in case of DBD. AB0-incompatible transplantations were performed only in living donations. A negative pre-transplantation cross-match was mandatory for all cases. If possible, grafts were transplanted in the right or left iliacal fossa with arterio-venous anastomoses to the external iliacal artery and vein, respectively. Ureteral anastomosis was modified according to Lich-Gregoire and stented using a double J-catheter. Double J-catheters were left in situ for six weeks and removed transurethrally. Until 2005, patients received a suprapubic catheter and a transurethral catheter for two weeks in order to achieve no pressure urinary diversion. After 2005, only transurethral catheters were placed for five days post transplantation. The usual immunosuppressive regime for AB0-compatible RTx-recipients included a triple therapy, consisting of tacrolimus, mycophenolate mofetil and steroids in reduced doses. Basiliximab or anti-thymozyte globulin were used as induction therapy. Use of cyclosporine A was abandoned in 2006. ganciclovir was used for CMV-prophylaxis depending on CMV status of donors and recipients. Cotrimoxazol was used for Pneumocystis carinii prophylaxis for 100 days following transplantation.Primary outcome for the first matched pair analysis was incidence of post-transplant malignancy . Primary outcome for the second matched pair analysis was 5-year-graft and -patient survival. Secondary outcome parameters included frequencies of delayed graft function , frequencies of primary nonfunction , episodes of biopsy proven acute rejection (BPAR) within one year after RTx and 1- and 5-year creatinine and eGFR .t-test. For continuous variables which are not normally distributed, median and quartiles  are given and a comparison between groups was performed with the Mann-Whitney U test. For categorical variables, the Fisher’s exact test was used. One and five-year patient survival, death-censored graft and overall graft survival were estimated by Kaplan-Meier methodology [p-values ≤ 0.05 were considered statistically noticeable. Cox proportional hazards regression models with univariable and multivariable logistic regression analyses of matched cohorts were used to determine independent factors influencing patient, death-censored, and overall graft survival at five years [p-value less than 0.05 in the likelihood ratio test) was performed for the final multivariable model. Results are shown as hazard ratios (HR) with 95% confidence interval (CI) and p-value of likelihood ratio test. All statistical analyses were performed with IBM SPSS® Statistics 24 for Windows .Normally distributed continuous variables are presented as mean ± standard deviation (SD) and groups were compared utilizing the student’s hodology  and compve years . Univarin = 36, 55.0%) of the PTM-RTx cohort was male (n = 10) were the two most common types of cancer, followed by gynecological malignancy (n = 7) (n = 17). Only one patient was diagnosed with a TNM-stage T4 prostate cancer pre transplantation. RTx was performed on average 105.6 ± 78.1 months after diagnosis of malignancy. Age at RTx was 62.5 ± 8.6 years and PTM-RTx-recipients had been on renal replacement therapy (RRT) for 60.6 ± 31.7 months . Majority of malignancy had been diagnosed pre RRT (44 before (67.7%); 21 during (32.3%) RRT). Patients suffering from gynecological cancers had the longest time of RRT, whereas patients with a history of thyroid cancer had the shortest time of RRT and oncological follow-up, respectively. PTM-RTx-recipients with history of urothelial tumors had the third longest time of RRT. Yet, in these cases, it is unknown if the urothelial carcinoma itself was the reason for RRT. Triple immunosuppressive therapy was used in the majority of cases consisting of steroids (95.4%), mycophenolic acid (93.8%) and tacrolimus (80.0%). Cyclosporine A (15.4%) and mTOR-inhibitors (1.5%) were only used in selected cases. Induction therapy was given in 54 (83.1%) cases . In the matched cohorts without history of PTM, induction therapy was given to 54 (83.1%) and 55 (83.3) patients, respectively.Between 1 January 2000 and 31 July 2012, 1217 patients received a RTx at our center. A total of 838 RTx patients met the inclusion criteria and were included in further analysis. Of these, 65 (8.0%) patients had a history of PTM. The average age of patients at the time of cancer diagnosis was 53.5 ± 10.9 years and the majority  . AssuminAfter 12 months, 88.1% of patients were still on steroid-based therapy combined with mycophenolic acid (78.0%) and tacrolimus (62.7%) or cyclosporine A (16.9%) while five patients were switched to an mTOR-inhibitor (8.5%). There was no difference in doses applied and drug levels targets between patients with and without a previous history of malignoma. In addition, frequency of induction therapy and used drugs was also similar between PMT-RTx and RTx patients. In case of BPAR, steroid pulse therapy was applied.To analyze the incidence of post-transplant malignancy, a matched pair analysis was conducted based on age (±5 years), sex and time under immunosuppressive drugs (±1 year). PTM-RTx and RTx recipients were matched 1:1 achieving similar results for age , sex  and time under immunosuppressive drugs  .When donor characteristics were compared, PTM-RTx- and RTx-donor cohorts did not differ noticeably regarding demographic factors  and frequencies of living donation or ESP allocation. In addition, cold and warm ischemia times as well as HLA matching were comparable . When ann = 9), but also lung (n = 2), breast (n = 1) and esophageal (n = 1) cancer. Nine (13.9%) RTx-recipients reported a history of de-novo malignancy after RTx. Leading malignancy among these were skin (n = 5) and one of the following: renal, lung, lymphoma and Barrett’s carcinoma. Kaplan-Meier analysis was used to display disease-free survival for patients with tumor recurrence, de-novo, and second de-novo malignancy  and 86.2% (RTx) of patients showed an uneventful post-transplantation course over the study period of 60 months without developing any form of malignancy . Howeverlignancy . Intereslignancy . In addip-value = 0.438)) or overall graft-survival . Kaplan-Meier analysis revealed comparable outcomes in 5-year patient, death-censored and overall graft survival. When analyzing the post-transplant graft-specific outcome, no differences were found regarding PNF, DGF, BPAR or one- and five-graft function (using eGFR as surrogate) or death censored graft loss  renal transplantation (RTx)-recipients and RTx-recipients. Survival rates of PTM-RTx-recipients and RTx-recipients with recurrence (red), second de-novo (blue) or de-novo (green) cancer after renal transplantation (RTx) were estimated by Kaplan-Meier methodologyUsing time under immunosuppressive therapy as matching criteria allows inclusion of one of the most established risk factors for developing cancer following SOT. However, it also entails a bias when analyzing patient and graft survival times. Therefore, a second 1:1-matched pair analysis was conducted to investigate patient and graft survival. Matching criteria were age, sex and underlying end-stage renal disease . There w2; RTx 26.9 ± 3.8 kg/m2). When graft specific outcomes were compared, frequencies of PNF, DGF, BPAR as well as one and five-year eGFR were similar (p = 0.1). Yet, when 5-year death-censored and overall graft survival were estimated, noticeable differences were revealed. Patients in the PTM-RTx cohort had a lower death-censored  as well as overall graft survival  five years after RTx. The leading cause of death-censored graft loss in both groups was rejection, with a comparable median time from RTx to event .When analyzing recipient characteristics , a highe similar . However similar . Kaplan- similar a, death- similar b and ove similar c stratifp-value = 0.007) as well as overall graft survival .Next, Cox proportional hazard regression models with univariable and multivariable logistic regression analysis were used to further investigate the influence of PTM on patient and graft survival. Unadjusted Cox proportional hazard modeling showed that PTM-RTx patients had a 4.198  as well as late (5-year) graft loss in PTM-RTx recipients in the Eurotransplant region.The prevalence of cancer is steadily rising in the general population as well as among RTx-recipients. Due to RTx-specific risk factors, including ESRD, RRT, viral infections, immunosuppressive therapy, as well as general risk factors , RTx-recipients suffer from an increased risk for malignancy ,18,19,20Most data on outcome from PTM-Tx recipients have been provided by large registry analyses. Israel Penn was first to build a transplantation registry, the Israel Penn International Transplant Tumor Registry, now Cincinnati Tumor Transplant Registry (CTTR). It reports recurrence rates of up to 21.0% and the development of secondary malignancy in approximately 33.0% of PTM-RTx-recipients, which is in line with more recent data from Acuna et al. ,10. BratThis study provides further evidence for inferior survival rates in PTM-RTx recipients by reporting worse five-year overall patient survival. Of interest, survival rates in the RTx cohort are comparable to larger cohorts, previously published from our center, in which 5-year overall survival rates between 87.1% and 92.4% were noted. (CITATION) This provides further evidence that choosing a highly selective cohort for the matched pair analysis did not introduce a bias in the analysis. In general, our survival data confirm previous reports indicating worse outcomes in terms of cancer mortality, all-cause mortality and outcome of posttransplant de novo malignancies in PTM-RTx-recipients ,9,24. BrRegarding graft specific outcomes, Livingston-Rosanoff et al. only recently used the UNOS database to analyze outcomes from RTx patients with PTM . In accoThe ever-widening disparity between supply and demand for transplantable grafts has led to ongoing discussions about when PTM patients should be considered eligible for RTx. Analyzing data from the US transplant registry over a 30 year period, Blosser et al. could not find changes in cancer-associated mortality in RTx-recipients despite advances in both immunosuppressive, but also cancer therapy, yet without including patients with a PTM history . BrattstOne advantage for using a case control matched-pair single center data in comparison to large registry data is the ability to provide a more sophisticated analysis on graft specific outcomes by providing granular analysis of associations. The current study provides evidence for inferior one- and five-year overall and death-censored graft survival for PTM-RTx-recipients. This was confirmed by uni- and multivariable analysis, demonstrating a direct correlation between PTM and graft loss. The reason for this remains unknown. One reason might be the acceptance of inferior grafts for PTM-patients. However, data here presented showed no noticeable difference regarding baseline donor characteristics (including KDPI and KDRI) or HLA-mismatches. Another reason for worse graft outcome may be subtle impairment of recipients caused by previous cancer-specific treatment in combination with the impact of RRT and waiting time on the immune system of RTx-recipients . HoweverMost studies that analyze Tx-recipients either compare them to the general population, thereby not taking immunosuppressive therapy and its influence into account, or to patients with HIV or other immunosuppressive diseases . In thesIn conclusion, this matched pair analysis based on retrospective single center data from a German transplant center within the ET region provides first evidence for inferior graft outcomes in RTx-recipients with PTM. While exact causes for an increased graft loss remain unknown, this study advocates focusing on meticulous graft monitoring in PTM patients in addition to a heightened surveillance for cancer recurrence or development of second de novo malignancy.


G-quadruplexes are believed to have important biological functions, so many small molecules have been screened or developed for targeting G-quadruplexes. However, it is still a major challenge to find molecules that recognize specific G-quadruplexes. Here, by using a combination of surface plasmon resonance, electrospray ionization mass spectrometry, circular dichroism, Western blot, luciferase assay, and reverse transcriptase stop assay, we observed a small molecule, namely, oxymatrine (OMT) that could selectively bind to the RNA G-quadruplex in 5′-untranslated regions (UTRs) of human vascular endothelial growth factor (hVEGF), but could not bind to other G-quadruplexes. OMT could selectively repress the translation of VEGF in cervical cancer cells. Furthermore, it could recognize VEGF RNA G-quadruplexes in special conformations. The results indicate that OMT may serve as a potentially special tool for studying the VEGF RNA G-quadruplex in cells and as a valuable scaffold for the design of ligands that recognize different G-quadruplexes. G-quadruplexes are four-stranded structures found in DNA/RNA sequences that are rich in guanine residues. Compared with double-helical DNAs, G-rich RNAs are relatively unconstrained and easily form G-quadruplexes . Both anRadix sophorae flavescentis, can efficiently repress the expression of VEGF and has an effective anticancerous effect (in vitro was confirmed and characterized through surface plasmon resonance (SPR), electrospray ionization mass spectrometry (ESI-MS), and circular dichroism (CD) melting studies. The results showed that OMT was a highly specific binder for the VEGF RNA G-quadruplex. No affinity of OMT was found with other G-quadruplexes or with DNA double strands. Furthermore, the results of luciferase assays confirmed the binding of OMT with the VEGF RNA G-quadruplex in cells. Western blot experiments showed that OMT selectively repressed the expression of the VEGF protein. SPR and luciferase experiments also showed that OMT can recognize the VEGF RNA G-quadruplexes in different conformations. Altogether, OMT showed recognizability for the VEGF RNA G-quadruplex. The results may provide a valuable scaffold for further developing the ligand binding–intended G-quadruplex with high specificity, which may serve as the specific inhibitor for VEGF.Accumulating evidence suggests that oxymatrine (OMT) , a naturs effect . Here, wTM, China. Detailed sequence information is described in TM), MgCl2, and dNTPs were then introduced into the reaction mixture. Then, the reaction mixtures with the reacting solutions were placed into an incubator . The reaction product (1 µL) was used as a template for the next quantitative PCR (qPCR) analysis. The primers for qPCR were designed as a forward primer [5′-GCTAGCTCGGGCCGGGAGGA-3′] with a reverse primer [5′-CATGGTTTCGGAGGCCCGACCGGG-3′] for the VEGF RNA G-quadruplex, and the forward primer [5′-TAATACGACTCACTATAGGG-3′] with the reverse primer [5′-CATCCTTCCCAGAGGAAAAGC-3′] for the BCL-2 RNA G-quadruplex template. Then, qPCR analysis was performed on a CFX ConnectTM detection system (BIO-RAD) with SYBR® Green. All ΔCt values were calculated by subtraction of the Ct value of qPCR with the compound from the Ct value without the compound.The sequences for reverse transcription were ordered and purchased from GenePharmaFor the RTase reaction-based stop assay, a reaction mixture of template RNA (0.3 µM), OMT (varying concentration) and the fluorescently labeled primer  was kept at 80°C for 3 minutes, followed by ReverTra Ace reverse transcriptase (TOYOBO), MgCl2, and dNTP introduction into the reacting solution at room temperature, and left for 30 min. The product was subsequently analyzed using 1.2% agarose gel.CD spectra/melting curves for G-quadruplexes were evaluated using the AVIV model 430 CD spectrophotometer. The samples  were purchased from Takara™ and GenePharma™. The oligonucleotides were maintained at 95°C for 5 minutes, followed by a gradual return to ambient temperature. The melting curves were obtained by monitoring at the 265/295-nm CD band. The CD spectra were recorded from 220 to 320 nm. The Tm value of each sample was calculated by the following method: ellipticity was plotted with a function of temperature and fitted in GraphPad Prism 7 software using a nonlinear sigmoidal dose-response model with a variable slope. The solutions for CD and UV spectra were prepared as 0.45-ml samples (10 μM) in the presence of KCl/NaCl (100 mM) with Tris buffer . Melting curves of duplex DNA analysis with or without OMT was performed on a Shimadzu model UV2600 UV–Vis spectrophotometer with a temperature controller. The melting curves were acquisitioned by monitoring at 265-nm band by scanning continuously from 40 to 75°C.TM, Canada). All 5′-biotin–labeled and annealed nucleotides (1000 RU) were purchased from TakaraTM and introduced into the running buffer . Nucleotides were immobilized on the biotin sensor chip surface using a biotin–streptavidin sensor chips kit obtained from NicoyalifeTM, Canada. Flow cell 1 served as the control surface, where no target oligonucleotides were captured. Diluted OMT solutions  were set up with the running buffer. The samples were then injected at a flow rate of 25 μL/ min during the association phase at 25°C. Data were fitted with TraceDrawer software.All binding studies based on the SPR phenomenon were performed on a two-channel open SPR instrument  mass spectra were measured, under the condition modified from the previous report, using an Exactive Orbitrap® mass spectrometer  in the negative ion mode. . Dataset−5 mbar .2+), dNTP (concentration: 200 μmol/L), and 5 U Pfu DNA polymerase. The PCR assembly was conducted as follows: denaturation at 95°C for 3 min followed by 30 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 30 s and termination by incubation at 72°C for 5 min. The product from the PCR reaction assembly was further amplified using primers P1 and P34 in a 50-µL reaction solution containing 0.5 µL of the PCR assembly mixture, 200 μmol/L of each dNTP solution, 0.5 μmol/L of each primer, 2.5 U Pfu DNA polymerase, and 10 × Pfu Buffer (containing 2 mmol/L Mg2+). PCR was conducted as follows: denaturation at 95°C for 3 min followed by 30 cycles at 94°C for 30 s, 55°C for 30 s, and 72°C for 30 s and termination by incubation at 72°C for 5 min. Fragment B was synthesized under the same conditions with the oligonucleotides P33 to P68 and amplified with the primers P33 and P68. Fragments A and B were purified using agarose gel. The vector pcDNA3.1(+) Invitrogen™ was digested with the enzymes BamHI and EcoRI (Fermentas). Fragments A and B were finally cloned into pcDNA3.1(+) using the ClonExpress® Entry One Step Cloning Kit. The resulting plasmid was named as pcDNA3.1(+)-Rluc-IRES-Fluc. Mutant plasmids were conducted using mutant sequences -Rluc-IRES-Fluc plasmid. Briefly, the two luciferase genes, namely, Renilla luciferase (RLuc) and firefly luciferase (FLuc), were controlled by the cytomegalovirus promoter (CMV) and separated only by the VEGF IRES sequence (nts 746–1038) . The resequences  in PCR a6 cells per well. On day 1, the cells were exposed to transfection vectors using the Lipofectamine™ 2000 Transfection Reagent , according to the manufacturer’s protocol. On day 2, such cells were co-incubated with OMT. Following 24 h of incubation, the cell cultures in each well were subjected to lysis, and activities of firefly/Renilla luciferases were gathered using the Dual-Luciferase® Reporter Assay System . The enzyme activities were normalized  accordingly.On Day 0, HeLa cultures were incubated on 6-well plates at 1.0 × 10TM Cambridge, United States; 1:10,000 dilution). The immunodetecting step was performed using the secondary antibody/ECL. Proteomic expression profiles were quantitatively analyzed through Odyssey (LI-COR™) or enhanced chemiluminescence (Pierce™).HeLa cultures were exposed to OMT for 2 days. Consequently, cell lysis employed Solarbio LIFE SCIENCES™ R0010® buffer (Estonia). The lysis solution was centrifuged at 12,000 × g/min (20 min/4°C), and the supernatant was extracted. The samples were segregated over 10% SDS-PAGE gel and consequently placed onto a PVDF membrane. Subsequently, membrane immunoblotting was performed, using specific antibodies for VEGF, BCL-2, and NRAS ; and specific antibodies for GAPDH  using the following primers: F (5′-TGCATTGGAGCCTTGCCTTG-3′); R (5′-CGGCTCACCGCCTCGGCTTG-3′) for VEGF and F (5′-GCACCGTCAAGGCTGAGAAC-3′); and R (5′-TGGTGAAGACGCCAGTGGA-3′) for GAPDH, the latter serving as the reference/normalization gene.HeLa cultures were exposed to OMT for 2 days. RNA extraction employed the TRIzol® kit (Transgen Biotech™). The reverse transcription step employed EasyScript® one-step gDNA removal and cDNA synthesis super mix. qRT-PCR runs were conducted over the CFX Connect® . The analyses employed GraphPad Prism® Software .HeLa cultures were exposed to varying OMT doses for 2 days, with OMT cytotoxicity being consequently evaluated by the use of the CellTiter-Glo1 Luminescent Cell Viability Assayin vitro RTase reaction-based stop assay. As shown in To test whether OMT can bind to G-quadruplexes, we used the reverse transcriptase (RTase) reaction-based method modified from a previous study to compare the binding of OMT with different G-quadruplexes  Katsuda. We mixeD value estimated as 3.75 × 10−5 M, whereas no SPR signal was observed for BCL-2 nor NRAS RNA G-quadruplexes  of the G-quadruplex measured by CD in the presence or absence of OMT showed that OMT increased the Tm of the VEGF RNA G-quadruplex by 3.6°C . In contThe ESI-MS is a useful method for the quantitative study of noncovalent complexes . Therefoin vitro, we next tested whether OMT can bind to the G-quadruplex and regulate the protein expression in cells. First, we prepared a dual-luciferase construct, in which the entire VEGF IRES was cloned between Renilla and firefly luciferases (Encouraged by the results of experiments iferases . Althoug50: 10.3 mM) compared with TMPyP4 (16.35 µM)   (Cheng a6.35 µM) . Such re6.35 µM) . Accordi6.35 µM) , the sliThe binding selectivity of OMT toward the VEGF RNA G-quadruplex leads to the possibility that OMT can recognize the different conformations of RNA G-quadruplexes. As previously reported, the VEGF RNA G-quadruplex is a kind of “tunable” G-quadruplex. With different combinations of G-tracts in the sequence segment, the 17-nt sequence can form G-quadruplexes in different conformations. Several mutations of G to U can force the sequence to adopt G-quadruplexes in specific conformations, and such G-quadruplexes exert different effects on VEGF IRES activity. Based on that, we generated a series of mutated sequences as previously reported  Morris . Then, wSPR experiments also presented similar results. As shown in The OMT structure is distinct from the previously characterized G-quadruplex ligands. It is nonplanar and non-aromatic. In some reported literatures, nonplanar and non-aromatic molecules are also described as potent G-quadruplex ligands, such as the steroid derivatives  and alkaG-quadruplexes are considered a potential biomedical target. Finding molecules that selectively bind to the intended G-quadruplex and exhibit special activity is required. Here, we discovered OMT as the first selective binder for the VEGF RNA G-quadruplex. Both results of CD, SPR, and luciferase assays and ESI-MS showed the ability of OMT in binding to the G-quadruplex structure, and results also showed that OMT could recognize the VEGF RNA G-quadruplex in different conformations. OMT showed very low cellular toxicity, which makes it potentially promising for the drug development process. OMT affected the expression of the VEGF protein with the mRNA level of VEGF unchanged, which indicates that the translation is controlled by interaction of the VEGF RNA G-quadruplex with OMT. Thus, OMT may serve as a special tool for understanding the VEGF RNA G-quadruplex in the cell and help in the design of the ligands that recognize different G-quadruplexes.


Tetradesmus obliquus microalgae for the production of antioxidant starch films as flexible packaging material. Starch was plasticized with glycerol and blended with 1 w% polyallylamine chosen as an agent to modify the film physical properties. The addition of polyallylamine improved film water stability and water vapor transmission rate as well as mechanical stiffness and tenacity. The dried Tetradesmus obliquus microalgae, which showed an EC50 value of 2.8 mg/mg DPPH , was then used as antioxidant filler. The addition of microalgae provided the films with good antioxidant activity, which increased with microalgae content increasing. To our knowledge, this is the first study reporting the development of sustainable bioactive packaging films composed of almost 100% starch, and follows the European union’s goals on plastics strategy concerning the promotion of bio-based, compostable plastics and the setting up of approaches to prevent food waste with a simple plastic packaging.This study combines the use of corn starch and  Specific EU policies include packaging recycling in a cost-effective manner (https://ec.europa.eu/environment/strategy/circular-economy-action-plan_en (accessed on 5 December 2021)) and the use of bio-based, biodegradable and compostable plastics (https://ec.europa.eu/environment/topics/plastics/bio-based-biodegradable-and-compostable-plastics_en (accessed on 5 December 2021)). In the last two decades, the EU action plans boosted a number of research studies focused on the development of biodegradable plastics from renewable sources in response to the increasing demand of eco-friendly packaging materials. Indeed, the packaging industry, which accounts for about 40% of the total world production of plastics, is now facing a major challenge, that is the replacement of traditional oil-based plastics, like polyethylene (PE), polypropylene (PP), polystyrene (PS) or polyethylene terephthalate (PET), with biodegradable plastics, possibly deriving from renewable resources. In order to replace the traditional plastics in either flexible or rigid packaging, biodegradable polymers should possess a series of requisites like: be light, inexpensive, recyclable, tough, strong and easy to process. Additional characteristics are required for food packaging, where polymers should also have proper barrier properties against permeation by water and waterborne microorganisms to protect food from spoilage and prolong product shelf-life. In this context, the European Committee of the Regions expressed a clear demand for “future research on the relation between packaging and food preservation and possible approaches to prevent food waste without the use of complex plastic packaging” [Plastic plays a key role in different sectors of the modern society, including in the food sector where plastic packaging helps food preservation, reduces transport cost and overall contributes to lower the environmental impact of food production . Besidesckaging” .Among natural biodegradable polymers, in the last two decades, starch has received considerable attention as food packaging material due to its renewability and low cost .The main biodegradable polymers under investigation as alternatives to synthetic oil-based plastics are polylactive (PLA), polyhydroxyalkanoates (PHAs) and starch. A notable feature of PHAs is their production from microbial anaerobic digestion process ,6. StarcInitially, dry granular starch (GS) was investigated as biodegradable filler for commodity plastics, especially low density polyethylene (LDPE) and linear low density polyethylene (LLDPE), either with or without processing aids . Basicalper se is already low [More recently, starch has started to be used as thermoplastic starch (TPS) that is obtained by heating water swollen starch granules up to 90–120 °C, so that the structure of H-bonded starch chains is disrupted, and granule crystallinity is progressively reduced. Following this, so called, gelatinization process , starch eady low ,17. In aeady low . The graeady low ,20. Howeeady low ,22.Blending with synthetic polymers can improve the physico-mechanical properties of TPS. Lately, LDPE/TPS blends, compatibilized with either maleic anhydride-grafted-PE or maleic anhydride-grafted-PP, have been proposed as sustainable alternative to pure polyethylene ,24. AlthTPS was also blended with different biodegradable polyesters such as polylactic acid , polycapw/w) was investigated by evaluating film transparency, water vapor transmission rate, water uptake, soluble fraction, contact angle and mechanical tensile tests.The present study aims at producing 100% TPS bioactive films with physico-mechanical properties suitable for packaging applications by combining the biodegradability of starch and the bioactivity of microalgae . The novTetradesmus obliquus microalgae  as film bioactive filler. Indeed, Tetradesmus obliquus microalgae is known to possesses chlorophylls, carotenoids  [T. obliquus was chosen for this study because it is a renewable source that can be easily produced at larger industrial scale to replace fossil derived chemicals with bio-based compounds. As compared to conventional plant sources, T. obliquus has the advantage to be cultivable on non-arable lands and with wastewater attaining biomass productivity up to 50–90 t/ha per year [T. obliquus as the best one in antioxidant properties when compared to other 40 microalgal strains [T. obliquus has such a relevant potential, its integration in plastic films was studied only in a single previous work [An additional novelty of the work is the use of dried arotene)  and phenarotene) . T. obliper year , almost  strains . Althougous work , while tMW(C6H10O5)n = 218 g/mol, 73% amilopectin and 27% amylose) , glycerol , poly hydrochloride  , 2,2-Diphenyl-1-picrylhydrazyl , methanol, Potassium nitrate , Sodium sulfate , Magnesium sulfate , Calcium chloride , Potassium phosphate dibasic , Sodium bicarbonate , Ferric sodium ethylenediaminetetraacetate , Ethylenediaminetetraacetic acid disodium salt dihydrate , Magnesium Chloride , Zinc sulfate , Cobalt chloride , Copper sulfate , Sodium molybdate  and 3-methyl-2-benzothiazolinone hydrazone (MBTH) were used as received.Corn Starch  used in the experiments is equipped with a Linkam HSF 91 thermoregulation device, which permitted to study starch gelatinization in the 25–90 °C temperature range.Tetradesmus obliquus, generally known as Scenedesmus obliquus and recently reclassified by Wynne and Hallan [3 10 mM; Na2SO4 0.7 mM; MgSO4·7H2O 1 mM; CaCl2·2H2O 0.5 mM; K2HPO4 2.5 mM; NaHCO3 10 mM; NaFeEDTA 28 μM; Na2EDTA·2H2O 80 μM; MnCl2·4H2O 19 μM; ZnSO4·7H2O 4 μM; CoCl2·6H2O 1.2 μM; CuSO4·5H2O 1.3 μM; Na2MoO4·2H2O 0.1 μM. The reactors were illuminated 24 h with 35 W led light lamps  supplying 200 µmol s−1 m−2 photons and fed with filtered (0.2 µm) 1 L/min air and 10 mL/min CO2. The cultivation was repeated in six batches. Initial biomass concentration was set at 0.3 g/L and biomass was harvested (5 min centrifugation at 3000× g) after 3 days when 2.4 ± 0.3 g/L biomass was attained. Harvested biomass was rinsed twice with distilled water and dried at 50 °C overnight. The resulting powder was reduced in size by grinding in an agata mortar and sieved through a 90 µm sieve. Biochemical characterizations were performed on the dry powder as follows. Proteins were determined as N% 6.25, based on N% determined with an elemental analyzer (EA 1110 CHNS/O). Starch was determined by using the total starch assay kit , following the protocol as described by de Jaeger et al. [2 flux. Pigments (chlorophylls + carotenoids) were extracted from 5 mg biomass with methanol at 70 °C for 10 min and quantified with the equation reported by Lichtenthaler [A strain of the microalgae d Hallan , isolated Hallan  was cultr et al. . Total cr et al. . Lipids r et al. , modifieenthaler . After mw/v starch suspension was submitted to a process of gelatinization at 90° for 30 min under magnetic stirring (500 rpm) and then to retrogradation by lowering temperature up to 40 °C. At this stage, additives were incorporated in the starch solution. Following 20 min of stirring at 40 °C, the solution (25 mL) was poured into a Teflon Petri dish (6 cm in diameter) and dried for 48 hr at room temperature to obtain thin films.Starch 2D-matrices (films) were prepared by the solvent casting and evaporation method. First, 2% Three sets of films were obtained according to the types of additive mixed with starch.w/w).Set 1: Starch films (S) containing glycerol (Gly) as plasticizer. Films were named S_GlyX where X stands for the amount of glycerol .Set 2: Starch films containing glycerol plus poly hydrochloride  as film property modifier (antiplasticization agent). Films were named S_PAH1_GlyX where X stands for the amount of glycerol .Set 3: Starch films containing glycerol (30%), poly hydrochloride (1%) and dried microalgae The whole microalgal biomass was used as filler in order to exploit the full bioactive property of microalgae biomolecules. Instead, the utilization of pure extracts would make loss some other biomolecules, synergic effects among biomolecules and it could increase their degradation rate.Thickness of the films was determined by a digital caliber, making three measurements on three distinct points for each film.−1 and co-adding 200 scans.Fourier-transform Infrared Spectroscopy (FTIR) was performed in Attenuated Total Reflection (ATR) by a Nicolet 6700  equipped with a Golden Gate ATR accessory (angle of incidence 45°), at a resolution of 4 cmThermal stability of starch films was evaluated by thermogravimetric analysis (TGA) by employing a Mettler TG 50 thermobalance. A thermal scanning interval of 25–500 °C and a scanning speed of 10 °C/min were set. Analyses were carried out under nitrogen flow, on 5–8 mg sample weight. The degradation temperature was determined in correspondence of the peak of the first derivative of the weight with respect to temperature.The ultraviolet and visible light barrier properties of the film were analyzed by a spectrophotometer HP DIODE ARRAY , following the ASTM D1746-09 standard procedure (Standard Test Method for Transparency of Plastic Sheeting). Measurements were carried out in absorbance mode at 2 nm intervals in the 190–820 nm range. The transparency of the films was calculated from the percent transmittance (T%) of light at 580 nm, according to the following equation:Three parallel experiments were performed for each film and data were reported as value ± standard deviation.2) and T is the film thickness (mm).Water vapor permeability tests were carried out following the ASTM method E96  with some modifications. Circular films were sealed as a patch onto a glass container (diameter = 1.4 cm and height = 5 cm) containing 10 mL of water. The weight of the assembled system was monitored over time, under constant temperature, and the water vapor transmission rate (WVTR) was determined according to the equation :WVTR  determination, 2 h-swollen films were dried at room temperature up to constant weight and weighted. SF was determined as:Three parallel experiments were performed.To determine surface wettability of starch films, the static contact angle was evaluated by the drop method. Particularly, a water droplet (Milli-Q-water) was deposited onto the film surface and a picture was captured. The resulting image was processed by Software Motic Image Plus 2.0 ML in order to determine the contact angle (CA) as follows:Mechanical features like Tensile Strength (TS), Young’s Modulus (E), Elongation at break (ε) and Tenacity (T) of films were determined by tensile tests by using an ISTRON 4502 instrument . For the analysis, films were cut into rectangular specimens (6 cm × 1 cm × 0.1 cm length × width × thickness) and fixed between the two Instron flat plates. Measurements were carried out at 100 mm/min deformation rate and with a 2 kN load cell.Tetradesmus obliquus microalgae as well as of the pure microalgae powder was evaluated by assessing the material ability to scavenge the DPPH  free radical [The antioxidant activity of films containing  radical . DPPH is0 is the absorbance of the initial DPPH solution while A is the absorbance of the solution after reaction with the antioxidant material.For each material, different concentrations, expressed as material weight (mg) per DPPH (mol), were assayed. Specifically, a weighted amount of sample, either polymer film or microalgae powder, was put in contact with a DPPH solution in methanol . After 30 min, the suspension was centrifuged, the supernatant was collected and submitted to UV-Vis spectroscopy. The amount of residual DPPH was determined by a previously obtained calibration curve (Abs vs. DPPH concentration), while the Scavenging Activity  for each sample amount was defined as:50) defined as the amount of compound able to decrease by 50% the initial DPPH concentration [The scavenging activity was then plotted vs. the antioxidant concentration (mg/DPPH mol) to determine the Efficient Concentration  is a polysaccharide deriving from different kinds of plants, from which is extracted in granular form. The size and morphology of starch granules strongly depends on the botanical source and may range from 2–3 μm to ca. 100 μm . The corUnder polarized light B, the MaIn order to set up the procedure for preparation of starch/additives compounds, the processes of starch gelatinization and retrogradation were followed by optical microscopy (OM) by heating the sample from 30 °C to 95° C at a 20 °C/min speed. In As it can be observed, gelatinization begins at ca. 75 °C and resulted to be complete at ca. 90 °C. By lowering temperature at 40 °C, set as the retrogradation temperature, granules’ recrystallization was not significant for the first 60 min. Therefore, the temperature of 90 °C was adopted for the starch gelatinization phase while 40 °C was chosen as the temperature for mixing the additives to the starch solution.w/w content (X); (2) the S_PAH1_GlyX series, containing 1% w/w PAH plus glycerol ; and (3) the S_PAH1_Gly30_Y series, containing 1% w/w PAH, 30% w/w Gly and dried microalgae Tetradesmus obliquus as bioactive filler at either 0.1, 0.5 or 1% content (Y). The obtained films were physico-chemically characterized in order to find out a correlation between their performance and the additive type and content.In order to investigate the effect of polyallylamine chloride (PAH) on the physico-mechanical features of glycerol-plasticized starch films as well as to endow the films with antioxidant properties, in this study, three sets of starch films were prepared: (1) the S_GlyX series, containing only glycerol (Gly) as plasticizer at either 10, 20 or 30% Flexible packaging films should be transparent and light in color to meet the aesthetic needs of consumers. However, light can accelerate photochemical degradation reactions that induce food deterioration . In genep < 0.05, In The introduction of 1% wt PAH in the films improved film transparency at all Gly contents b. HigherTetradesmus obliquus microalgae, the transparency was studied only for the S_PAH1_Gly30 film, containing 1% PAH and 30% Gly, because this film was shown to possess the best physico-mechanical properties as will be widely described later on. As can be observed in As far as the starch films containing the −1 is associated with the free hydroxyl groups, whereas peaks in the range 3000–2800 cm−1 are associated to the C-H stretching and the peak at 1639 cm−1 is attributed to the bending of the bond (H-O-H) of the absorbed water.−1, the bands related to C-O-C, and C-O-H vibrations are present while at 1334 cm−1 the bending vibrations of the bond (C-O-H) are observed [In the spectral region between 1200 and 900 cmobserved .−1 region, as a result of starch interactions with glycerol or PAH. Specifically, around 1000 cm−1, three bands are present in the starch spectrum at 1042 cm−1 (shoulder), 1015 cm−1 (shoulder) and at 995 cm−1 (peak). When glycerol was added in the films, the peak at 995 cm−1, related to C-OH stretching, was shifted to 998 cm−1  present in the films.Very small differences in the profile shape of peaks can be observed in the starch fingerprint, and particularly in the 1100–900 cm S_PAH1, , may be Thermogravimetric analysis (TGA) was carried out to determine the thermal stability of the different starch formulations and thus gaining information about the establishment of potential interactions between starch and additives. In the thermogram of the native starch film, a first weight loss of 10% is observable at around 100 °C related to evaporation of absorbed water and a second weight loss at ca. 300 °C related to polymer chain degradation. The presence of Gly at all of the explored contents did not affect significantly starch thermal stability, although a weight loss in the 200–250 °C temperature range is present in the S_Gly20 and S_Gly30 samples A, presumThe possibility of establishing strong interactions between plasticizers and an auxiliary component has been already explored in the literature for several purposes, like the control of drug release in pharmaceutical formulations  or preseThe study of film behavior in water environment is very important for packaging application ,63.In As far as films containing PAH are concerned, the non-plasticized sample (S_PAH1) shows a very low soluble fraction, even lower than the pure starch film, related to the antiplasticization effect of PAH consisting in the formation of strong H-bonds between PAH NH-groups and starch OH groups . When glWater swelling measurements and contact angle determination also evidenced a peculiar behavior of the S_PAH1_Gly30 film. Indeed, this film showed the lowest equilibrium swelling percentage 90%,  and the Water vapor permeability is an important feature for food packaging application. Usually, low permeability values are required for packaging films in order to reduce the transfer of moisture from the environment to food and to ensure longer storage times . HoweverGas transmission through a packaging material can take place mainly by two mechanisms: (i) pore effect, for which the gas crosses the material through small holes or cracks of the structure; (ii) solubility-diffusion effect, for which gas transmission is related to the concentration gradient between the two packaging sides and the gas solubility in the material. Gas diffusion depends on the characteristics of both the penetrating molecule  and the packaging material  . The pre2 day [2 day for S_GlyX films and 55–77 g/m2 day for S_PAH1_GlyX films. Therefore, the use of S_PAH1_GlyX films should be limited for the packaging of fresh foods characterized by high transpiration rate such as strawberries, asparagus, mushrooms [2 day target water vapor transmission rate of packaging materials has been suggested to avoid condensation of water vapor inside the packaging [In general, our films showed WVTR values higher than LDPE, that is the most used polymer in flexible packaging. Indeed, LDPE WVTR values range from 6.7 and 8.7 g/m2 day  while inushrooms , for whiackaging . Indeed,ackaging .The effects of glycerol and PAH on the mechanical properties of the films were investigated by performing tensile tests. In As for as the S_Gly series is concerned A, a sligInterestingly, the S_PAH1_Gly30 sample not only had a ductility higher than S_Gly30, but also showed greater Young modulus, strength at break and tenacity , highligT. obliquus microalgae as antioxidant filler.The S_PAH1_Gly30 film has the tensile strength (4 MPa), elastic modulus (60 MPa) and elongation at break (ca. 80%) lower than LDPE   but of tIn the Both the plasticizer glycerol and the anti-plasticizer PAH can establish hydrogen bonds with the starch in replacement of the water-starch H-bonding and starch-starch inter- and intra-molecular H-bond interactions. However, the strength of the PAH-starch H-bonding was hypothesized to be higher than that established between starch and glycerol, based on the electronegativity of the pair of acceptor and donor involved in H-bonding. Indeed, if starch is the acceptor of the hydrogen, with either C-O-C glycosidic linkage or a hydroxyl group, the hydrogen-donating effect of the amine group in PAH is higher than that of glycerol or water. On the contrary, if starch is the H-donor, with is OH groups, the electronegativity of the PAH nitrogen is lower than that of oxygen in glycerol. Therefore, in the competition of H-bonding with starch, glycerol is always weaker than PAH, thus justifying the higher tenacity of the S_PAH_Gly films with respect to the S_Gly ones . 2 and light levels, which could induce photoinhibition. T. obliquus is a fast-growing species able to grow under high light intensity with low sensitivity to photoinhibition [T. obliquus contains different bioactive molecules such as pigments (carotenoids and chlorophylls), phenols, proteins, polysaccharides and fatty acids [T. obliquus antioxidant and antiviral properties [T. obliquus was used in this study as antioxidant filler for the S_PAH1_Gly30 film. T. obliquus was produced from cultivation under optimal environmental conditions, for inducing high growth rate, high photosynthetic yield and high content of pigments and phenols [Microalgae are photosynthetic microorganisms often exposed to conditions inducing oxidant stress during growth, as high Ohibition . To facety acids . These boperties . For the phenols . Biomass phenols ,76. The  phenols .Expect starch (8%) and ash (1–3%), all the other microalgal components are well known to have antioxidant and other bioactive properties  ,77. In oT. obliquus microalgae are reported. As it can be seen, the introduction of microalgae did not significantly affect the film soluble fraction  corresponding to 2.2 mg/mL. Unfortunately, it’s difficult to compare the obtained EC50 value with literature data. Indeed, the antioxidant activity of T. obliquus (Scenedesmus obliquus), reported by Guedes and colleagues [karyotic . The higlleagues , ranged Heterochlorella luteoviridis and Dunaliella tertiolecta biomass in the starch film reduced proportionally the oxidation rate of oil during storage [Once entrapped in the S_PAH1_Gly30 film, the microalga was able to maintain its activity, as shown in  storage . The sta50 value of the microalgae entrapped in the film was determined by plotting the film radical scavenging activity vs. the amount (mg) of microalgae present in the film normalized for DPPH amount (mmol)  B.50 value of 1252 mg/mmol DPPH very similar to the EC50 value it showed when tested as a free powder. This finding suggests a good interaction of the DPPH radical with the microalgae when entrapped in the starch film, which is presumably related to a good dispersion of the microalgae in the film itself. These good results are promising in the view of the exploitation of T. obliquus also for other bioactive properties, as the antiviral activity, which is worth of investigation in future studies.Interestingly, when entrapped in the starch film, the microalgae showed an ECT. obliquus biomass as bioactive filler endowed the films with a remarkable antioxidant activity, which increased proportionally to the microalgae content (up to 1%), maintaining at the same time good physico-mechanical properties of the film. T. obliquus biomass is a source of several bioactive compounds, easily producible at large industrial scale, without the needing for arable lands. Specific biodegradability tests were not carried out in this study. However, we expect a biodegradability behavior similar to the native starch, or even higher when the microalga is introduced in the film due to the presence of nitrogen, which can act as a nutrient for microorganisms. This study has shown how its antioxidant potential can be exploited in the production of a bioactive starch-based films. The obtained films hold the potential to be used as flexible packaging.In this study, an approach to obtain bioactive 100% TPS flexible films with physico-mechanical properties as much as closer to LDPE has been set up. The use of a polymeric amine, polyallylamine, in a very low content (1%) in glycerol-plasticized starch films permitted to improve film water stability, transparency and water vapor transpirability as well as to increase the film mechanical resistance and tenacity. The starch film with 30% Gly content and PAH 1% exhibited an elastic modulus and tensile strength of the same order of magnitude of LPDE and a good elongation at break even if lower than LDPE. The introduction of the


This article presents the results of finite element analyses of the influence of reaction forces on stresses and strains at the contact points of the rollers of prism supports with cylindrical surfaces of the main journals of large-sized crankshafts. The analyses of strains and stresses, as well as the depth of their occurrences, in the case of the shaft journal and support rollers were carried out using Hertz contact theory and the finite element method. These calculation results proved to be highly consistent. Additionally, they provide a basis for stating that, in the case under consideration, permanent deformations do not significantly affect the values of the measured geometrical deviations nor the profile forms of the supported main crankshaft journals. A crankshaft is responsible for the correct operation of an engine and the conversion of linear motion into rotary motion . TherefoAfter a crankshaft has been manufactured, it should be properly verified to ensure that it meets the geometrical specification of the product. One of the commonly used methods of fixing and supporting cylindrical machine elements during measurements, is to support them in prisms . This meIn the case of fixing in prisms, unambiguous determination of the reference element is troublesome due to the shaft displacements that occur during its rotation . These dZhang et al.  presenteAmor et al.  presenteAidoudi and Bessrour  presenteSaha and Jackson  proposedAn et al.  presenteWen and Tang  presenteWang et al.  presenteA review of the literature shows that considering the roughness of contact surfaces is of great importance. Therefore, this approach was used in this article, and the influence of the contact between the shaft journal and the rollers of the measuring prisms on the results of crankshaft journal roundness measurements was examined. The article presents finite element analyses of the stresses and deformations of a rough shaft journal in contact with the smooth rollers of prisms made of steel and bronze. The obtained results were compared with analogous analyses for a smooth shaft journal. The influence of the shaft weight and contact deformations on the results of roundness measurements were examined using an outlined analysis.This paper is structured as follows: Depending on the adopted support conditions, during the measurement of shafts, deflections and elastic deformations occur. These issues are of particular importance in the case of measuring large-sized, slender shafts, such as the crankshafts of marine engines. Depending on the adopted support conditions, deflections and elastic deformations of the shaft can affect the results of measurements in terms of geometric quantities. This makes evaluation of the geometry of the shaft in these conditions unreliable. An effective method in eliminating deflections and elastic deformations of shafts is the use of a measurement system equipped with a so-called flexible support for the measured object. Such a system is presented in One of the most important elements of the proposed system is the elastic support. The purpose of the elastic support is to implement variable values of reaction forces at the contact of rollers with the main journals during crankshaft rotation. The implemented values of the reaction forces guarantee zero deflection values on the individual supported main journals. The rollers are mounted on pneumatic cylinders and a force sensor is placed between the actuator and the rollers. The actuators are equipped with precise current-controlled valves, cooperating in a feedback system with force sensors, thanks to their implementation, variable reaction forces can be performed in a controlled manner.The support heads are constructed in the form of roller prisms, as such, there are minimum values of frictional resistance at the contact points between the rollers with the journals, corresponding to the rolling friction resistance. The resistances depend on the value of the forces occurring at the contact between the rollers with the pin, and thus on the value of the force activated by the support. The problem of surface pressures and stresses is directly related to the problem of forces and frictional resistance at the roller–journal contact. By analyzing the impact of frictional resistance on the results of measurements of geometric quantities, it was found that the impact was negligible. However, a separate problem is the influence of surface pressures.These pressures may cause, not only elastic deformation of the material of the pin or roller, but also a permanent deformation of cooperating elements. From the point of view of the measurement technique, including measurements of the deviations and shape contours, the influence of this factor on the measurement of geometrical quantities cannot be ignored and it should be individually analyzed, depending on the parameters of the measuring system and those of the measured object.As emphasized earlier, a feature of the presented measurement system is that it allows to eliminate the elastic deformations of the crankshaft, thanks to the support system which realizes variable reaction forces that guarantee zero deflection values on the supported main journals. Therefore, it is justified to carry out an analysis of the influence of reaction forces on elastic or permanent deformations in the contact places of the rollers of the prism supports with the cylindrical surface of the supported main pins.Considering the above, the required values of the reaction forces were determined for the adopted test object—a crankshaft from a medium-speed main drive engine from a Buckau Wolf R8 DV136 ship, which is 3630 mm long and weighs 9280 N, with ten main journals with diameters of 149 mm and eight crank journals with a 144-mm diameter. The determination of the reaction forces was carried out using the finite element method.Modeling was carried out using Midas 2019  . The finNext, a gravity load was applied to the model. An analysis consisting in the determination of shaft deflections and reaction forces acting on a supported shaft  was performed using the linear static Nastran solver SOL101 ,25). The. The25])Based on the model, calculations were made for the reaction forces, guaranteeing the elimination of the deflections on the main journals; a graphical presentation of this is shown in According to According to the diagram presented in E = 210 GPa and Poisson ratio ν = 0.28. In the second case, the shaft was made of steel with the above-mentioned material properties, while the support roller was made of aluminum–bronze with a Young’s modulus E = 210 GPa and Poisson ratio ν = 0.34.Next, the Hertz formulas  were useThe calculated stress values for the shaft journal were: in the case of a steel shaft cooperating with rollers of prisms made of steel, 215.2 MPa, while in the case of a steel shaft cooperating with rollers made of bronze, 175.3 MPa.The presented results of the calculations of stresses and the depth of their occurrence are correct in terms of the linearity of stresses as a function of elongations (the applicability of Hooke’s law).To build a reliable finite element model for the analysis of contact phenomena, the model had to demonstrate the compliance of the obtained calculation results with the results obtained from theoretical formulas in the field of linearity. Obtaining this compliance was the basis for switching to calculations in the nonlinear range, where permanent deformations appear. The necessity to take finite element analysis of the contact between the surfaces of the shaft journal and the support roller into account required the modeling of elements to the centers of rotation of the shaft and the support roller .The finite element model for contact analysis was build using eight-node six-sided isoparametric finite elements, and CPENTA six-node five-sided isoparametric elements. The model results consisted of 58,184 finite elements and 351,474 degrees of freedom. The shaft modeling part consisted of 44,312 finite elements and that of the roller consisted of 13,872 finite elements. In the contact area of the shaft and the support roller, a refined mesh was used to accurately reflect the values of the maximum contact stresses and the depths of their occurrences . The resR = 1253 N was determined (using non-linear Nastran solver—SOL106) [Based on the built finite element model, the case of the shaft journal and support roller made from steel using the values of the maximum stresses and the depths of their occurrence with the reaction force P0 is determined as follows:meanP is:b is given by:r is given by:E′ is defined as follows:Considering the analyzed case from the point of view of the Hertz contact theory, it can be reduced to the contact analysis of two cylinders; according to which, Hertz contact stress Based on this, it is possible to determine the values and locations of the maximum shear stresses and the maximum von Misses stresses. The detailed procedure for determining these values is presented in .A comparison of the results of the finite element analyses and the Hertz contact theory is presented in R = 1253 N.R = 1253 N.R = 1253 N are shown in Finite element calculation results, showing reduced stresses in the shaft and support roller according to the von Mises hypothesis for the case of a steel shaft journal and a bronze support roller, under the action of a reaction force of E = 210 GPa, while the yield stress was eR = 250 GPa and the Poisson ratio was ν = 0.28. In terms of elasticity, the material tensile diagram is rectilinear with a steep inclination angle, the tangent value of which, in the adopted reference frame, corresponds to a Young’s modulus of E = 210 GPa. The steeply sloping waveform is valid until:As stated earlier, for the steel of the shaft the Young’s modulus was Above the yield point, the tensile diagram is still rectilinear, but with a smaller inclination angle, the tangent of which, in the adopted reference frame, corresponds to a value of 100 MPa .R = 150 N, “zero” position;Stage 1—initial shaft pressure on roller the with a force of R = 4000 N;Stage 2—rotation of the shaft by an angle of +4.5°, a gradual increase in the pressure force to a value of R = 4000 N, “zero” position;Stage 3—rotation of the shaft by an angle of −4.5° under the pressure of the force R = 4000 N;Stage 4—rotation of the shaft by a further angle of −4.5° under the pressure of the force R = 0 N.Stage 5—rotation of the shaft by an angle of +4.5° and a gradual decrease in the pressure force to Research in the field of non-linearity included a simulation of contact cooperation during rolling of the shaft journal over the support roller for a distance equal to 5 mm, which corresponded to an angular rotation of the roller equal to approximately 9°. Since the assumed initial value of the reaction force was equal to 1253 N, there were no permanent deformations and reaction force R was increased to 4000 N (simulating a larger or more slender shaft). The calculations were carried out in five successive stages :Stage 1—With these stages, the cycle of turning the shaft journal “back and forth” over the supporting roller over a distance of 5 mm was carried out. The calculations with the use of the non-linear Nastran solver—SOL106 required 5 iterations in stage 1 and the 50 iterations in the remaining stages. The change in the angular position of the shaft was simulated using a prescribed displacement, which, according to the adopted boundary conditions, resulted in shaft mesh rotation.The “turning” procedures were carried out for a shaft journal with an ideal circle profile (not burdened by irregularities in the shape outline), and a shaft journal with a real profile, with an irregular roundness outline. These procedures were carried out, as previously mentioned, for the cooperation of the shaft journal and the steel supporting roller, and the steel shaft journal and the supporting roller made of aluminum–bronze.Therefore, considering that there are no permanent deformations in the support roller, the stress analyses were limited to residual stresses occurring in the surface layer of the shaft. Of course, not only were the stress values analyzed, but the permanent deformations were analyzed as well. R = 4000 N, permanent stresses remained in the surface layer of the shaft material. The distribution of these stresses is shown in After turning the steel journal with no irregularities over the steel support roller with a force of R = 4000 N. Permanent stresses remain in the surface layer of the shaft material, the distributions of which are shown in A similar form of permanent stress was shown in the results of the analyses for the case of the cooperation of the steel shaft journal, which is not burdened with irregularities in the profile, with the bronze support roller with a force of In the case of a shaft journal with an irregular outline, the actual profile of journal No. 10 was selected for the analyses of permanent stresses and deformations. The actual measured profile of journal No. 10 was recorded during 1024 measurements with equidistant points located on its circumference. The detailed procedure and description of the SAJD measurement system can be found in . The SAJThis profile was evenly distributed over a length of 471 mm, which corresponded to the circumference of a wheel with the nominal diameter of 150 mm for the measured main journal .x = 200 mm. This coordinate determined the maximum height of the measured profile, equal to 0.035 µm. An enlarged fragment of this selected section of the real profile, 6 mm long (from −3 mm to 3 mm), was transformed into the new coordinate of the peak occurrence, x = 0 mm, and was presented against the background of the shaft profile with no outline irregularities (shown in In the tests of permanent stresses and deformations of the actual profile, they were limited to a segment of the actual shaft profile, near the contact with the support roller around coordinate shown in .The distribution of the permanent stress values in the surface layer of the steel shaft journal with the actual profile of journal No. 10, after this profile has been turned over the support roller made of aluminum–bronze shown in .R = 4000 N, for the journal profile not burdened with outline irregularities, and the actual one after turning these contours on support rollers made of steel and of aluminum–bronze are shown.A cumulative comparative diagram of the reduced stresses according to von Misses, remaining in the shaft journal made of steel as a function of the depth of their occurrence is shown in This article presents the results of analyses on the influence of reaction forces on the stresses and strains at the contact points of the rollers of prism supports with cylindrical surfaces of the main journals of a large-sized crankshaft.The finite element models used in the research were verified using the Hertz contact theory in terms of the values and locations of the maximum stresses, and showed a high consistency. Comparing the accuracy of the developed models to those presented in the literature, and also verified based on the Hertz contact theory, it can be deduced that similar results were achieved ,32,33.Then, based on the verified finite element model, nonlinear static analyses was carried out, including the rolling of smooth and irregular shaft journals over steel and bronze rollers. The results of the analyses showed that rolling a shaft with irregularities on the steel support roller caused large permanent deformations on the surface of the shaft journal that were almost twice as compared to the case of rolling on a roller made of aluminum–bronze. This also applied to turning of the shaft journal, not burdened with outline irregularities . The maxThe presented comparative assessment showed that the material used to make the roller has a large impact on the value of the permanent stresses; this is very visible in From the point of view of the aim of the undertaken research, there is no concern that, during the measurements of the tested objects, permanent deformations may occur because of the movement cooperation of the rollers of the prism supports with the supported crankshaft journals. Thus, it can be assumed that, in the case under consideration, permanent deformations do not significantly affect the values of the measured geometrical deviations and the outlines of the shapes of the main journals of the crankshaft. Therefore, it can be concluded that the aim of the study has been achieved.Further research, based on the model presented in this article, will include comprehensive modeling of the system presented in


FEBS Open Bio celebrates its 10th birthday. To celebrate the journal's first decade, we present this special anniversary issue, comprised of editorials, reviews, and research articles especially commissioned for the occasion. In this introductory editorial, we invite the reader to join us as we reminisce over the journal's past, celebrate its present, and look forward to its future.This month,  To celebrate the journal’s first decade, we present this special anniversary issue, comprised of editorials, reviews and research articles especially commissioned for the occasion. This issue has been a year in the making, and we are sure there is something to interest everyone in its virtual pages. But first, please join us as we reminisce over the journal’s past, celebrate its present and look forward to its future. FEBS Open Bio celebrates its 10th birthday. From publishing only four articles in its first volume in 2011 to publishing over 260 articles in 2021, the journal has come a very long way. Many factors have contributed to the journal’s success, including the authors who choose to submit their work, the academic editors and reviewers who generously contribute their time and expertise, the readers who download, share and build on the work, and of course, the tireless efforts and commitment of the editorial office staff, the FEBS Publication Committee, the FEBS administrative staff and our publishing partners, Elsevier (2011–2015) and Wiley (2016–). The greatest contributing factors to the journal’s success are the dedication, experience and knowledge of the journal’s original Executive Editor, Mary Purton, who for eight years managed and guided the journal from its very beginnings to a recognised and respected journal and who continues to oversee FEBS Open Bio and its companion journals as FEBS Press publisher.This month, The FEBS Journal is now 54 years old , FEBS Letters is a bit younger, at 53, and Molecular Oncology is now 14. FEBS Open Bio was FEBS’ first fully open‐access journal. We are proud to be part of this illustrious publishing heritage and fully subscribe to the FEBS Press motto of ‘science publishing by scientists’: all submissions to FEBS Open Bio are considered by editors who are active scientists and are reviewed by researchers working in the field of study.The journal is the youngest of four siblings: To celebrate the journal’s first decade, we present this special anniversary issue, comprised of editorials, reviews and research articles especially commissioned for the occasion. This issue has been a year in the making, and we are sure there is something to interest everyone in its virtual pages. But first, please join us as we reminisce over the journal’s past, celebrate its present and look forward to its future.Following the move to digital publication at the end of the 20th century, the movement towards open‐access publishing began to gain momentum at the start of the 21st century. By 2011, recognising that the move towards open access could not be ignored, the Federation of European Biochemical Societies (FEBS) set up a fully open‐access and online‐only journal. Further details of the journal’s genesis are provided in the following editorial [The FEBS Journal (Richard Perham), FEBS Letters (Felix Wieland) and Molecular Oncology (Julio Celis).The first issue of the journal was published in December 2011, with Mary Purton appointed as Executive Editor. Initially, most articles were transferred from other FEBS Press journals, but direct submissions were also received. In those early days, the journal benefitted from the expertise and support of the Editors in Chief of The FEBS Journal and FEBS Letters to form FEBS Press, allowing the journals to be better consolidated with the other activities of FEBS. Molecular Oncology transferred a year later , finally bringing all four journals together under the FEBS Press banner.In 2016, the journal transferred to Wiley, joining In 2017, the journal started a new Education section, for the publication of research on biochemistry education. This section, which is edited by Angel Herráez and Luciane V. Mello, was introduced to encourage research into innovative teaching methods for biochemistry and the dissemination of best practice. The publication costs of all Education articles published in the journal are covered by FEBS, and so, there is no cost to either author or reader. At the time of writing, 16 Education articles have been published in this ‘journal within a journal’, many of which have been well‐read and cited. We are proud to be the venue for many excellent and freely accessible education articles, as effective training of the next generation of biochemists and molecular biologists is essential for the future of research.st January 2020 [As the journal settled in with its new publisher, direct submissions continued to grow and the editorial office support was expanded with the appointment of Kerry Dresser as Editorial Assistant in 2017 and Duncan Wright as Editorial Associate in 2018. Jacob Weller replaced Kerry in 2018. By 2019, it was apparent that the time had come to separate the role of Executive Editor into two distinct positions, an academic Editor‐in‐Chief to set journal strategy and a full‐time Editorial Manager to handle the day‐to‐day operations of the journal. In 2019, the FEBS Publications Committee appointed Miguel A. De la Rosa as the journal’s first Editor‐in‐Chief, with his term commencing on 1Under Miguel’s Editorship, the journal has continued to grow, not only in terms of number of submissions and acceptances but also in visibility and variety of journal content. Downloads of articles published in the journal continue to increase dramatically year by year: downloads from the journal website alone (not including downloads of the articles from other sources) increased by a third from 2019 to 2020, and by the end of August 2021, downloads were already 6% greater than in the total of 2020, suggesting that articles published in the journal are useful to the community.Miguel has also implemented several new initiatives to improve journal performance and diversify its content, which are summarised below.FEBS Open Bio was first founded, its editorial board was comprised entirely of members of the editorial boards of the other FEBS Press journals; many of these original editors are still with the journal, and we are exceedingly grateful for their long service and dedication. As submissions to the journal increased, it soon became apparent that more editors were needed, to handle not only a greater quantity of manuscripts but to also ensure we had the expertise to handle manuscripts on diverse subjects. In addition, we have considered the geographic spread and gender balance of the board in our appointments, to help ensure that our editors best reflect our authors and readers. The editorial board was expanded in 2013 and 2018, and in 2020 and 2021, we appointed fifteen and two new editors, respectively. We are very pleased that all of our new appointments have assimilated well into the journal environment, and we look forward to continuing to work with them for many years.When FEBS Open Bio and Molecular Oncology. Ruzhica has brought many years of postdoctoral experience to the role; she completed her PhD at the University of Freiburg, followed by postdoctoral positions at University Medical Center Freiburg and subsequently the German Cancer Research Center (DKFZ). Ruzhica quickly became an integral part of both the journal teams, spearheading new initiatives and drawing on her experience and network to promote the journals. We are exceptionally grateful to Ruzhica for her initiative, professionalism and attention to detail, which has elevated the journal. We have also recently appointed Ioannis Tsagakis as a second Assistant Editor working across both FEBS Open Bio and Molecular Oncology; Ioannis completed a very successful internship at FEBS Press last year, at which time he impressed everyone with his diligence and enthusiasm.At the beginning of 2021, Ruzhica Bogeska was appointed as Assistant Editor working across When it comes to journal performance, our top priority is to maintain thorough and timely peer review of all submissions. To ensure that we always have willing, available and expert reviewers, we introduced a new Editorial Advisory Board (EAB) in 2020. The members of our EAB are committed to providing assistance to our editors, through offering independent evaluations when reviewers disagree, providing thorough peer review at short notice and ensuring that authors have appropriately revised their manuscript when the original reviewer is no longer available to check. At present, the EAB is comprised of 34 members, and we hope this number will increase in future to better reflect the broad scope of the journal.With the journal regularly receiving over a hundred submissions a month, it is important that we have access to a large pool of dedicated and experienced reviewers with diverse research interests. While the EAB is very valuable in this regard, its numbers are too few to review the majority of submissions to the journal. To meet the growing need for reliable reviewers, we reinvigorated our volunteer reviewer pool in 2020 by inviting new members, ensuring that our records on reviewers’ research interests and contact details were current and encouraging our editors to make full use of our volunteers. We continue to publish an annual Reviewer Acknowledgement in December to thank all of our reviewers, and this year is no exception – the Acknowledgement is included at the end of this issue.The journal had its first editorial board meeting in October 2020 – while we had all been looking forward to meeting each other in Seville and enjoying a drink in the warm weather, fate had other plans. Once it had become apparent that there was to be no travel that year, the decision was made to have a virtual meeting and postpone the in‐person meeting to a later date. We did find that the virtual meeting had its boons – it was far better attended than could be expected of a physical meeting, with editors attending from all over the world and not just Europe. We consider this editorial board meeting a major milestone for the journal, and it provided a sense of community, involvement and journal ownership to our editorial board.The journal has also started to publish special ‘In the Limelight’ issues, which include review articles focused on a topic of especial interest. Our first ‘In the Limelight’ section focussed on the important issue of Bioplastics and was commissioned to accompany the FEBS Special Session on Science & Society – Plastics: revolution, pollution and substitution at the 45th FEBS Congress, which at the time was scheduled for July 2020. We fully expected that the special section would materialise after the Congress, little expecting that the event would be postponed a year and then transformed into a virtual event. As it happened, the Bioplastics section preceded the Science and Society session by four months, and we hope it was able to stimulate interest in the event .The reviews for the ‘Membraneless organelles’ section were also commissioned to tie in with a symposium to be held in Seville in March 2020, which was postponed and then unfortunately cancelled. Despite this setback, the silver lining was that the accompanying issue was published in September of this year. The three review articles in this issue highlight very different factors that are involved in the condensation of membraneless organelles . We are To present articles published in the journal to a lay audience, we started a new series of articles called ‘Insights’ in 2021. This series, coordinated by our editor Cornelia de Moor, aims to present findings with broad significance to a general audience. In these Insight articles, the main findings are summarised and presented within a wider context. To date, we have published four Insight articles, two of which were written by postdocs interested in science communication, while the other two were written by editorial office staff. We consider it very important for the journal to be involved in outreach activities and are delighted that this series has had such a productive start in its first year. We invite any researchers interested in contributing to this series to contact the editorial office.th anniversary of the journal was an ideal time to give our editors a platform to discuss their research and other topics of interest. Our editor Beáta Vertessy agreed to help coordinate this series, dubbed ‘An open chat with…’, and to date, we have published two interviews, with more ready to be published in 2022.We are extremely proud of our prestigious Editorial Board. Although they are the central pillars of the journal, their contributions are largely ‘behind the scenes’ and not visible to the reader. We decided that the 10FEBS Open Bio prizes for the Speed Talks presented at the FEBS Congress. At this year’s 45th FEBS Virtual Congress, a jury comprised of members of the local organising committee and the journal’s editorial office selected two excellent Speed Talks, the presenters of which won €200 each. The winners were:Lígia Fão of University of Coimbra, Portugal, for the speed talk ‘Altered autophagy‐dependent c‐Src/Fyn degradation in Huntington’s disease – impact on NMDAR activity’.Natalia Kruglova of the Institute of Gene Biology RAS, Moscow, Russia, for the speed talk ‘Cell‐to‐cell transmission of SARS‐CoV‐2 is resistant to neutralizing antibodies’.Since 2018, the journal has published the talk and poster abstracts for the annual FEBS Congress as a special supplement issue. Building on this tradition, Miguel has introduced new initiatives to strengthen the ties between the journal and the Congress, as well create ties with the annual Young Scientists’ Forum (YSF), which precedes the Congress. The first of these new developments was the introduction of two Martin Toul of Masaryk University, Brno, Czech Republic, for the poster ‘Engineering protein dynamics for the understanding of divergent evolution of the Renilla luciferase’.In addition, the journal continues the tradition of awarding a prize to the presenter of an excellent poster; this year’s FEBS Open Bio Poster Prize was awarded to:The 20th FEBS YSF, originally planned for Lovran, Croatia, in 2020, and then held as an online event in 2021, brought  together about 100 PhD students and postdoctoral researchers, to present their research, discuss their experiences, participate in workshops on career skills  and attend lectures by distinguished speakers. At this year’s event, the winners of the best short oral presentation and best poster received discounts on the article processing charge for future submissions to the journal. The winners were as follows:Best short oral presentation (SOP) prize: Angela Garcia‐Mato of the Instituto de Investigaciones Biomédicas ‘Alberto Sols’ (IIBM), Madrid, Spain, for the talk ‘IGF‐1 via PI3K/AKT activation promotes survival and anabolic metabolism in HEI‐OC1 auditory cells’.Best poster prize: Carmen Escalona‐Noguero of IIMDEA‐Nanoscience, Madrid, Spain, for the poster ‘Nanoparticle‐mediated delivery of Cpf1 for the generation of improved gene editing tools’.We would like to take this opportunity to congratulate the winners above as well as the winners of the Congress daily poster prizes and wish them all the best with their future careers.FEBS Open Bio Article Prize, which is awarded to an early‐career researcher who has authored (either as first or corresponding author) an article of especial interest published in the journal in a 12‐month period from July to June of the previous year.This year also saw the introduction of the FEBS Open Bio Article Prize was awarded to the paper’s first author, Arpit Katiyar, who is now a postdoctoral researcher at SGPGI Lucknow, India. Arpit has been invited to submit an abstract on his current work for presentation at the 46th FEBS Congress to be held in Lisbon, Portugal, 9–14 July 2022. In addition, Arpit has also created a video describing the award‐winning work, which can be viewed here: https://febs.onlinelibrary.wiley.com/journal/22115463/prize‐winnerA jury of three members of the journal’s Editorial Board, Stuart Ferguson (Oxford), Takashi Gojobori (Mishima) and Alex Wlodawer (Frederick), selected the following as the winning paper: ‘HSF1 is required for induction of mitochondrial chaperones during the mitochondrial unfolded protein response’, authored by Arpit Katiyar, Mitsuaki Fujimoto, Ke Tan, Ai Kurashima, Pratibha Srivastava, Mariko Okada, Ryosuke Takii and Akira Nakai of Yamaguchi University School of Medicine, Ube, Japan . In thisCongratulations to Arpit, and we look forward to seeing you in Lisbon!th anniversary between the journal’s management team and Mary Purton. Miguel suggested that we mark the occasion in some way and Duncan proposed an entire issue dedicated to the 10th anniversary, with review content and interviews. We invited members of our editorial board to submit review articles and invited both editors and the authors of highly cited articles previously published in the journal to submit their latest research for consideration of inclusion in the special issue.The idea for the special issue emerged in October 2020, following discussion of the upcoming 10th anniversary, but also reflect the journal’s strong history of publishing sound science. Eventually, we decided that this history should be represented by the most cited article in the journal’s history: ‘Extracellular vesicle‐mediated transfer of long non‐coding RNA ROR modulates chemosensitivity in human hepatocellular cancer’ [To mark this special occasion, we knew that we had to have a fantastic cover image. It had to somehow represent the 10 cancer’ . This arth anniversary issue is an editorial by Pierre Cosson, a long‐standing member of our Editorial Board, on his experiences developing a curriculum for a new Bachelor‐Master course at the University of Geneva [We are very excited about the final issue line‐up. In addition to the aforementioned editorial on the journal’s history , the artf Geneva . This isth anniversary. The variety of exciting and cutting‐edge research topics in this issue reflects the diversity of our Editorial Board, and we are sure these articles will be of interest to our readership. Two review articles in this issue focus on neuronal function, from two different angles: biochemistry and phylogeny. In the first article, Sandro Sonnino and colleagues at the University of Milano discuss the beginnings of the elucidation of a ganglioside ‘code’ which may regulate the physiological function of neurons [We were delighted at how many members of our Editorial Board agreed to submit review articles for the 10 neurons . This re neurons . As physA fortuitously timed inclusion to this anniversary issue is Maria F. Drincovich’s review of metabolic diversity and reconfiguration in peach fruit  – while Autophagy is integral for cell function, and we are pleased to include two review articles on different aspects of this multifaceted process. Mitophagy is a specialised form of autophagy that primarily removes damaged and dysfunctional mitochondria, but it may also play a role in the development of specialised cell types through an independent mechanism. In the first of our review articles on autophagy, Ivana Novak and colleagues at the University of Split, Croatia, describe the role of the mitophagy receptor BNIP3L/NIX in the development of certain cell types and outline the gaps in our knowledge of the mechanisms involved . This miThe above case study highlights the importance of combined computational and experimental methods in the study of pathogens, and this is further emphasised in two excellent minireviews that focus on specific viral and bacterial proteins, respectively , 15. In We are also very pleased to introduce the four research articles that were commissioned especially for this issue. Tushar Patel, the author of the most highly cited article in the journal’s history  , reportsc through site‐directed mutagenesis [c, furthering our understanding of its extramitochondrial roles.We are also delighted to be able to include two exciting research articles from members of our Editorial Board. Dietmar Manstein and his colleague Simon Hennig of Hannover Medical School submitted an article demonstrating that combining quantum dot triexciton imaging (QDTI) with laser scanning microscopy can increase lateral and axial resolution . This meagenesis . MimeticFEBS Open Bio in the future because they anticipate a smooth, friendly publishing experience, and want their article to be free to read immediately on publication in a journal that is committed to maintaining the integrity of the publication record.Finally, we would like to thank everyone involved in the success of the journal. The journal is the sum total of its published articles, its editorial board and all the time and expertise contributed by authors, reviewers, editors, editorial office staff and the staff of our publishing partners. It is our hope that increasing numbers of authors will choose to publish their work in The former and current FEBS Secretary Generals, Israel Pecht and Vaclav Paces; the former and current FEBS Treasurers, Iain Mowbray, Alan Fersht and Frank Michelangeli; the former and current Publications Committee Chairs, Félix M. Goñi, László Fésüs and Johannes Buchner; and all other Executive and Publications Committee members past and present, for their significant help with the journal’s launch and success.All the members of our Editorial Board past and present, who regularly commit their time and energies to ensure the success of the journal;All the reviewers who kindly gave of their time and expertise to ensure fair and thorough peer review;Everyone who has submitted their work to the journal over the past decade;All former and current members of the FEBS Press Editorial Office, the FEBS administrative staff and all the support staff at both Elsevier and Wiley, for their hard work and encouragement.Very special thanks go to Mary Purton, as without her commitment, experience and many years of service, the journal would not be the success it is today. We hope that we can continue to maintain the very high standards Mary set for the journal, and it is an honour to continue this legacy.We would like to extend our gratitude to the following:th anniversary issue and will continue to support us during the next decade.And finally, we thank you, our loyal readers! We hope you enjoy this 10