As COVID-19 gradually shifts to an endemic state, stakeholders throughout academia have started settling in with an emerging “new normal” paradigm for science, dominated by the virtual environment. Journals make no exception to this phenomenon and are now striving to keep up with a myriad of developments in many fields, partly due to enhanced global focus on science. Despite pandemic-related hardship, the Journal of Medicine and Life (JML) has continued to grow, receiving significantly more and higher quality submissions from researchers worldwide.To accommodate the heightened interest for dissemination in JML, as well as a strong incentive coming from the academic community to promote content at better standards in general, our editorial board and ownership has committed to an ambitious reform pathway, which we hope will rise JML to a top-tier publication in the coming years. In this editorial, we are uncovering exciting developments coming into effect starting this first issue of 2021, hoping to ignite interest in the journal and invest in working with us to disseminate your work.First and foremost, we would like to present some information about this publication to readers who are not aware of the background of JML. It was founded as a peer-reviewed journal in 2008 and has published over 1.200 pieces of original work addressing topics from various fields of medicine and life sciences, including original research, systematic reviews, special reports, case presentations, major medical breakthroughs, and expert communications. We have also been continuously advocating for an educational and community-building environment by ensuring designated sections that inform our readers about exciting international congresses, teaching courses, and other relevant academic events.Despite having reached some important indexing milestones , we are now looking to expand our reach toward new horizons by preparing to apply for inclusion in Clarivate Analytics Web of Science databases.In doing so, we have started a profound reorganization of the journal, addressing several important facets. The most striking change is our brand-new visual identity, which had already been rolled out within our website during last year. Starting this first issue of 2021, we have also given our print edition a modern, fresh look, matching our authors’ immersive content. To support efforts on this front and improve the overall quality of journal output, our editorial team has also been expanded with several consultants .Moreover, we have decided to increase our publication frequency from four to six issues per year (bi-monthly). We hope this will attract potential authors looking for a shorter average duration from submission to publication. The Journal of Medicine and Life will continue to be provided under the Creative Commons license, supported by our fees, which have now been consolidated under a single article processing charge, change which we hope will incentivize high-quality submissions to the journal. A crucial planned area of intervention is also consolidating and updating our editorial board, which will develop a brand-new, attractive editorial process that will enable the journal to reach the highest impact in the coming years.https://www.theneurotech.eu). The NeurotechEU Alliance, bringing together eight leading universities and 250+ partners from all regions of Europe, aims to build a trans-European network of excellence in brain research and technologies to increase the competitiveness of European education, research, economy and society. In the upcoming 2021 issues, we plan to showcase our partners’ contributions to the broader topic of widening access and participation in the higher education arena.Finally, we would like to highlight our recent partnership with NeurotechEU – the European University of Brain and Technology (office@medandlife.org.We thank our readership and authorship for your most valuable contribution in supporting us come thus far and genuinely hope you will consider joining our efforts to bring this new chapter of JML’s existence to its full potential. If you are interested in joining our efforts in any way possible, please write us a letter at


An essential effect of environmental stiffness on biological processes in cells at present is generally accepted. An increase in arterial stiffness with advanced age has been reported in many publications. The aim of the present review is to summarize current information about possible chemical reactions and physical processes that lead to tissue stiffening and result in age-related diseases in order to find methods that can prevent or retard time-dependent tissue stiffening. The analysis of published data shows that bound water acts as a plasticizer of biological tissues, a decrease in bound water content results in an increase in biological tissue stiffness, and increased tissue stiffness leads to NF-kB activation and triggered actin polymerization—NF-kB activation is associated with age-related diseases. It can be suggested that changes in bound water content through changing tissue stiffness can affect cellular processes and the development of pathologies related to aging. Both age-related diseases and COVID-19 may be associated with tight-junction disruption and increased tissue stiffness and permeability. Proteins and other macromolecules in living tissue are functional provided that they contain water molecules as an integral part. Biological macromolecules are inactive in the absence of bound water. The changes in bound water and free water ratios have been associated with age-related and protein-conformational diseases and have an important role in time-dependent processes in biological tissues ,2,3. TheThe behavior of unfrozen water below the equilibrium free water freezing temperature has been discussed by Wolfe and co-authors . Free waIt has been concluded that water in close contact with macromolecules is no longer liquid but somewhat structured . Bound wThe effect of living tissue plasticization by water on tissue stiffness is typically neglected in the current state of the art. The aim of this paper is to review mechanisms that demonstrate how changes in bound water content can lead to changes in tissue stiffness, promoting the aging process and triggering related pathologies.Discher and co-authors reported in 2005 that cells respond to the rigidity of their planar substrate . It has It is important to take into account that mechanical properties depend on the content and state of water bound to biological macromolecules ,3. The bThus, it is very important to prevent or retard the transformation of intracellular tightly bound water into extracellular loosely bound and free water in order to mitigate an increase in tissue stiffness.It has been shown that tissue stiffness increases as a result of tissue crosslinking by advanced glycation end products (AGEs). Dehydration and glycation may be considered as subsequent steps during the aging process ,25. A deA number of AGE inhibitors that are effective in decreasing AGE accumulation in tissues have been proposed, such as aminoguanidine, pyridoxamine, aspirin, ALT-711 , thiamine, tenilsetam (antidementia drug), and others ,34,35,36Polyphenols have been suggested as inhibitors of AGEs ,40,41,42Higher flavonoid consumption has been associated with a lower arterial stiffness. Isoflavones and anthocyanins have been recommended as the most efficient for the improvement of vascular health . PolypheIn an in-vitro study, it was shown that bovine coronavirus infections can be neutralized by theaflavins extracted from black tea . Ueda etA more hydrated and intact intervertebral disc tissue has been observed in rats who consumed an omega-3 fatty acids diet . At the Arterial wall stiffness is related to cardiovascular diseases . For exaAntihypertensive drugs, such as angiotensin-converting enzyme inhibitors, have been suggested to decrease arterial stiffness ,61. The It can be expected that AGE content will increase in the presence of high-molecular-weight chitosan . HoweverN-(2-hydroxypropyl)-3-trimethylammonium chitosan chloride (HTCC) was proposed as an inhibitor of human coronavirus HCoV-NL63. It was suggested that HTCC blocks human coronavirus interaction with the cellular receptor, angiotensin-converting enzyme type 2 (ACE2) proteins [proteins .Association between vitamin D deficiency and arterial stiffness determined from pulse wave velocity measurements was reported in a number of publications. Vitamin D deficiency was associated with the increase in arterial stiffness in children with type 1 and type 2 diabetes and with chronical kidney disease as well as in the elderly population ,73,74,75Generally, the results from randomized controlled trials have been inconsistent and more large population studies should be conducted. It was reported that high-dose—but not low-dose—vitamin D supplementation was able to decrease arterial stiffness . The decIt has been recently reported that vitamin D supplementation could reduce the risk of death from the current COVID-19 epidemic. It is commonly accepted that vitamin D deficiency weakens the immune system. Vitamin D deficiency also increases tissue stiffness, and maintaining tight junctions was mentioned as one of mechanisms of vitamin D function . MaintaiVitamin D intake is very low in Italy, Spain and France . The mosVitamin E prevented an increase in femoral artery stiffness in diabetic Wistar rats . It was It was suggested that supplementation with vitamin D and vitamin E may increase resistance to novel coronavirus SARS-CoV-2 .Vitamin C decreases arterial stiffness ,86. A coVitamin K2 decreases arterial stiffness by preventing and reversing calcification of arteries and inhibition of inflammation in the vascular wall ,88,89,90It was reported that chitosan oligosaccharide ascorbate corrected deficiency in vitamin A, E, C and B groups .It was observed that salt consumption increases arterial stiffness and the risk of cardiovascular diseases ,93,94,95Optimal hydration retards development of age-related diseases . IncreasDehydration activates NF-κB . DehydraMatrix stiffening promotes F-actin polymerization regulated by small GTPase RhoA, Rho GTPases control cell proliferation and cytoskeleton remodeling and they are commonly hyper-expressed in tumors. Y-27632 also suppresses formation of stress fibers in cultured cells and decreases hypertension in several hypertensive rat models ,110,111.It is very important that children have an extremely low rate of hospitalization presented by the U.S. Department of Health and Human Services in It has been also reported that obesity as an underlying condition was observed in 48.3% COVID-19 patients. It was earlier demonstrated that obesity results in increased arterial stiffness .Strong dependence on a patient’s age allows us to conclude that current methods of treatment of age-related diseases have to be verified, as well for application in the treatment of COVID-19. Thus, COVID-19 is unique and differs from previous epidemics by the fact that children are very rarely infected by novel coronavirus . COVID-1Changes in water status lead to the change in extracellular matrix stiffness. The age-related decrease in tightly bound water content due to reduction of binding forces between water molecules and biological macromolecules results in the development of age-related and protein-conformational diseases due to an increase in tissue stiffness and permeability. It has been recently demonstrated that the ratio of extracellular water/intracellular water increases with advanced age and especially accelerates after the age of 70 years; a high value of this ratio is a major risk indicator for all-cause mortality.The role of bound water in biological processes is underestimated in the current state of the art. It is evident that taking into account the effect of bound water on protein conformations and stability can facilitate understanding mechanisms of biophysical processes in age-related diseases and protein-conformational diseases. It is more correct to say that bound water is essential to life than to say that liquid water is essential to life.Release of tightly bound water triggers glycation reactions that have a very important role in the aging process. Correlation between arterial stiffness and tightly bound water mobility can be expected. An increase in arterial stiffness is commonly accompanied by a decrease in glycocalyx coverage.The relationship of bound water with tissue stiffness has scarcely been studied for the time being and can be suggested for in-depth study in future.Biochemical glycation reactions and biophysical changes in hydration status are equally important in the development of physiological processes during aging. The possibility to regulate the hydration status of extracellular matrix could help in the development of new, more effective therapeutics for treatment of age-related and protein-conformational diseases.NF-κB activation was associated with matrix stiffening and the development of age-related diseases. Rho GTPases control cell proliferation and cytoskeleton remodeling, and the inhibition of RhoA/ROCK inhibits stress fiber formation and has the potential to decrease the stiffness of extracellular matrix and, in such a way, to treat a wide number of pathologies. In order to understand Rho GTPase inhibitors’ functional properties more clearly, it would be reasonable—in parallel with tissue stiffness—to study changes in the content of tightly bound water using nuclear magnetic resonance.Polyphenols, omega-3 fatty acids, vitamins D, E, C and K2, and statins have been suggested as de-stiffening agents. Research on tightly bound water’s role in de-stiffening is still in its infancy and needs a lot more attention. This review is focused on the role of dehydration and stiffness development in aging tissues. Evidently, other processes—for example, those involved in water–salt metabolism—have an important role in aging, and they could be discussed in special reviews.


Liver transplantation is the only curative treatment option for end-stage liver disease; however, its use remains limited due to a shortage of suitable organs. In recent years, ex vivo liver machine perfusion has been introduced to liver transplantation, as a means to expand the donor organ pool.To present a systematic review of prospective clinical studies on ex vivo liver machine perfusion, in order to assess current applications and highlight future directions.ClinicalTrials.gov registry was performed.A systematic literature search of both PubMed and ISI web of science databases as well as the clinicaltrials.gov registry, and these focus on use of ex vivo perfusion in extended criteria donors and declined organs.Twenty-one articles on prospective clinical trials on ex vivo liver machine perfusion were identified. Out of these, eight reported on hypothermic, eleven on normothermic, and two on sequential perfusion. These trials have demonstrated the safety and feasibility of ex vivo liver machine perfusion in both standard and expanded criteria donors. Currently, there are twelve studies enrolled in the Ex vivo liver machine perfusion seems to be a suitable strategy to expand the donor pool for liver transplantation and holds promise as a platform for reconditioning diseased organs. Liver transplantation is the only curative treatment option for acute liver failure and advanced hepatobiliary malignancies. Unfortunately, this therapy is limited by the supply of suitable donor livers for transplantation , as reflTo ameliorate the gap between supply and demand for liver transplantation, the use of marginal livers has increased –10. MargHistorically, liver machine perfusion had been considered for organ preservation, but early approaches were not widely adopted. Recently, with much technological progress being made, there is a renewed interest to implement this technique to not only assess, but also to improve the quality of liver grafts Fig. 35–37].–37.35–37As such, one of the primary goals of ex vivo liver machine perfusion is to mitigate the effects of graft IRI , 39. ThrA systematic literature review was performed following the Preferred Reporting Items for Systematic reviews and Meta-Analyses for Protocols 2015 (PRISMA-P) . In case of discordance, the corresponding author NR was consulted, and consensus was made via discussion. During stage one of data extraction, the titles and abstracts of all retrieved records were reviewed and unsuitable studies were excluded. During stage two, full text articles of remaining studies were read carefully and assessed for inclusion criteria. For the identification of clinical trials, the study description was reviewed by NR and MP and results discussed with the remaining authors. Extracted data were reviewed and analyzed by all authors.Systematic literature search of the National Library of Medicine database and the ISI Web of Science database identified 1.956 unique records. Based on to title and abstract, 1.887 papers could be removed, leaving 69 articles for full text analysis. Of these, 48 publications did not meet inclusion criteria. Two of the identified publications were video-articles and therefore excluded. One publication using hypothermic oxygenated perfusion (HOPE) was removed given that its primary focus was on the evaluation of the safety and feasibility of normothermic regional perfusion prior to HOPE in DCD donors with extended warm ischemic times rather than the assessment of this technique for the purpose of preservation. Further five studies were identified as follow-up studies on perfusate, liver or bile duct samples of pre-published clinical trials and consequently excluded. Taken together, 21 publications on prospective clinical trials using ex vivo liver machine perfusion for graft preservation or graft assessment in liver transplantation were identified Fig. . Eight rClinicalTrials.gov identified a total number of 22 trials, out of which 12 met the inclusion criteria . One-year patient and graft survival were 90% in both groups, with no death related to graft function. However, the HMP group had significantly shorter hospital length of stay (HLS) (p = 0.006), and significantly lower levels of post-operative peak aspartate aminotransferase (AST) (p = 0.011), peak alanine aminotransferase (ALT) (p = 0.044), total bilirubin (p = 0.042), and serum creatinine (p = 0.013) ) that no differences in clinical outcome such as post-transplant death or severe complications following machine perfusion could be identified. Raigani et al. have demonstrated that steatotic liver grafts suffer from deficits in antioxidant capacity, efficient energy utilization, and lipid metabolism during normothermic liver machine perfusion. . Thus, aWhen comparing hypothermic and normothermic machine perfusion approaches, it is important to note that normothermic systems enable metabolic characterization and viability assessment of the grafts by “simple” read out parameters like bile production, bile pH, or lactate clearance, while reduced metabolic activity during hypothermia restricts the viability assessment capacity of HMP systems. Although specific surrogate parameters or biomarkers might be able to fill this gap, as shown by Muller et al. , normothLastly, as success in transplantation remains dependent on multidisciplinary care, decreasing strain on treatment teams remains important. By allowing for prolonged preservation periods up to 30 h , NEVLP hIn line with our conclusions, the Italian Society of Organ and Tissue Transplantation (SITO) recently released their evidence-based position paper on machine perfusion in liver transplantation , recommeIn conclusion, ex vivo liver machine perfusion has significant potential to revolutionize the field of liver transplantation. In a field challenged by a persistent organ shortage and continued donor demographic changes threatening the quality of the already scarce pool of available grafts, the use of this technology may help in addressing the daunting waitlist mortality. Within its first decade of clinical introduction, it has proven to be safe and feasible, for hypothermic and normothermic perfusate temperatures, and demonstrated improved early graft function, compared with SCS preserved grafts. These findings have allowed reintroduction of DCD programs in countries with long mandatory no touch periods and salvage of grafts that otherwise would have been discarded. Considering the early stage of the technology, it remains critical to gain further insight into the underlying mechanisms that lead to observed clinical change, and the possibility of expanding use of this technique for treatment and reconditioning of otherwise unusable donor grafts.


Hopea hainanensis, a Dipterocarpaceae tree species dominant in lowland rainforests in Hainan (China) but endangered due to anthropogenic disturbances, is dispersed mainly by wind because of its sepal-winged samara. Here, we measured dispersal-related intraspecific samara traits of H. hainanensis, and analyzed their variation and correlation in relation to plant height, DBH (diameter at breast height), and elevation plant location. Great variations in the samara traits existed, and the variations were larger within than among individuals, which indicated a “bet-hedging” strategy of this species. Plant height, DBH, and elevation explained slight variation in the samara traits. Samara dispersal potential is mainly affected by the samara mass and morphological traits. Samara settling velocity was significantly positively correlated with fruit mass, seed mass, length and width, as well as samara wing loading, and negatively correlated with wing mass ratio, wing area, and wing aspect ratio. Substantial proportions of intraspecific variation in samara dispersal are explained by the samara mass and morphological traits. Natural regeneration with human-aided dispersal is necessary for recovering the H. hainanensis population. This finding contributes to the generalization of trait-based plant ecology, modeling of seed dispersal in tropical forests, and conservation and recovery of rare and endangered species such as H. hainanensis.Propagule dispersal is a crucial life history stage, which affects population recruitment and regeneration as well as community structure and functions. The windborne process of samara dispersal is affected not only by samara traits and other plant traits, but also by environmental factors. Therefore, studying samara traits related to its dispersal and intraspecific variation in relation to other plant traits and environmental factors could help to understand population distribution and dynamics.  Plant functional traits are morphological, physiological, and phenological characteristics by which plants interact with their environment during evolutionary processes. Such traits link ecological processes on multiple scales, from individual, population, community, and ecosystem to landscape . Trait-bRare and endangered plant species usually have populations composed of few individuals restricted to local communities and ecosystems. Based on the mass ratio hypothesis , communiPropagule  dispersal is one of the most important stages in plant life history . As sessSamara  is present in 25 orders, 45 families, and 140 genera of angiosperms . It contDue to evolutionary adaptation, seed trait tradeoffs , such asHopea hainanensis Merr. et Chun, is a Dipterocarpaceae species distributed in the northern tropic (Hainan in China and Nghe An in Vietnam). Its samaras (seeds) dispersed mainly by wind due to its sepal-winged samaras. It was once a dominant species in lowland tropical rainforests; nevertheless, it is currently listed as an endangered species in IUCN  What are intraspecific variations of samara traits? (2) Are such traits related to intrinsic or extrinsic factors? (3) Is there any tradeoff among samara traits in H. hainanensis? and (4) Which samara traits affect samara settling velocity in H. hainanensis? in IUCN  and firs in IUCN . Currentmination , habitatmination , and popmination . AlthougHopea hainanensis is an evergreen tree, ca. 25 m in height, naturally distributed in valley and windless lower foothills at 300–900 m a.s.l. in lowland rainforests. A samara of H. hainanensis is composed of an ovoid main body (seed covered by pericarp hereinafter referred to as seed) and two sepal-wings oppositely attached to the seed  is the main distribution region of  forests .H. hainanensis trees from natural populations in Bawangling Nature Reserve. A total of 3,207 samaras were collected, including 876 samaras (51.5 ± 3.9 samaras per trees in average) with intact wings randomly collected from each tree. For the 17 trees, we also measured DBH , height (13–33 m), and elevation of locality (270–840 m. a.s.l.). All fresh samples were carefully numbered and transported to the laboratory as soon as possible for further measurements.We sampled ripe samaras from 17 mature, healthy, and high seed-setting rate Fmass, g), then each intact samara was used to determine samara settling velocity  and seed mass , respectively. Seed length  and width  were measured using a Vernier caliper. Detached-wing length , width , and area  were assessed using the WinFOLIA Leaf Analysis Software . To reduce variations in weight and morphology caused by seed desiccation, all measurements were completed within 1 week after collection.After measurements of SSV, samara wings and seeds  were carefully separated and weighed to determine wing mass  was calculated as SL/SW. Wing aspect ratio was calculated using WL/WW. Wing loading (WL) was calculated by dividing the samara’s mass by its wing area  was determined using ing area . To expl R 4.0.2 .H. hainanensis showed substantial intraspecific variations, both among and within individuals , WMR , and wing area , showing weak but significant negative correlation with DBH; and wing loading showed weak, significant positive correlation with DBH . The relationships between samara traits and tree height showed almost the same trends as those of samara traits and DBH, except regarding wing area ratio, which showed a weak positive correlation ; whereas SSV showed a weak negative correlation  with tree height , nor between SSV and SMI . SSV was positively correlated with fruit and seed mass, SL and seed width, and wing loading, but negatively correlated with WMR, wing area and aspect ratio  and extrinsic  factors . In thisnsis age . It is dnsis age . Interspnsis age . HoweverDacrydium pierrei in this area  or extrinsic (elevation) factors we explored in this study. Other factors such as genetic and edaphic factors need to be considered and might explain those intraspecific variations in future.Two contrasting hypotheses have been used to explain the relationship between seed traits and elevation – the “stress-tolerance” hypothesis and the “energy constraints” hypothesis . The “sthis area . In thishis area  for H. hH. hainanensis is highly affected by samara traits, which may have profound implications for population demography and genetics of H. hainanensis.Seed dispersal is a mechanism that allows plants to cope with environmental change, stochasticity, and uncertainty . Seed trH. hainanensis may have better seedling performance but inferior dispersal, as found in other species   . AnotherH. hainanensis. The findings in our study might be applied to other populations only with caution due to potential population genetic divergence. As such, future studies on samara trait variations of H. hainanensis among multiple populations are needed. Another limitation is that we only studied SSV in still air; the flight behavior and dispersal distance of H. hainanensis samaras would be more complicated in windy environments. Thus, field investigation of the spatial distribution pattern of seed rain and seed germination characteristics and wind-tunnel experiments  and extrinsic (elevation) factors could explain little regarding variations in samara traits. We verified that the samara dispersal potential of H. hainanensis was mainly affected by its mass and morphological traits.Substantial intraspecific  variations of samara traits in The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Y-BS and MD contributed to the study conception and design. Y-BS and X-LS-T collected the data. All authors wrote and reviewed the manuscript and have read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


We observed 794 predicted SARS-CoV-2 epitopes of which 52% were private and 48% were public. Ninety-nine percent of the public epitopes were shared with SARS-CoV and 5.4% were shared with either one of four common coronaviruses, 229E, HKU1, NL63, and OC43. Moreover, to assess the potential risk of self-reactivity and/or diminished T cell response for peptides identical or highly similar to the host, we identified predicted epitopes with high sequence similarity with human proteome. Lastly, we compared predicted epitopes from coronaviruses with epitopes from influenza virus deposited in IEDB, and found only a small number of peptides with limited potential for cross-reactivity between the two virus families. We believe our comprehensive in silico profile of private and public epitopes across coronaviruses would facilitate design of vaccines, and provide insights into the presence of pre-existing coronavirus-specific memory CD8+ T cells that may influence immune responses against SARS-CoV-2.While individuals infected with coronavirus disease 2019 (COVID-19) manifested a broad range in susceptibility and severity to the disease, the pre-existing immune memory to related pathogens cross-reactive against SARS-CoV-2 can influence the disease outcome in COVID-19. Here, we investigated the potential extent of T cell cross-reactivity against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that can be conferred by other coronaviruses and influenza virus, and generated an  Faced by unprecedent health and economic crisis from the coronavirus disease 2019 (COVID-19), the scientific community is pushing forward with efforts to develop vaccines and treatments to mitigate its impact. While the severity of symptoms have been reported to be associated with age, gender, and comorbidities such as cardiovascular diseases and chronic respiratory diseases , 4, and As immune memory by related pathogens has shown to help reduce severity and spread of the diseases –7, pre-eThe strains infecting humans belong to either alpha and beta genera of coronavirus. The alphacoronavirus contains human coronavirus 229E (HCoV-229E) and HCoV-NL63, while the betacoronavirus contains HCoV-OC43, and HCoV-HKU1, Middle East respiratory syndrome coronavirus (MERS-CoV), SARS-CoV, and SARS-CoV-2 . It is kin silico profile of public and private predicted epitopes. We also expanded the map of cross-reactivity from exact matching peptides to those with a high sequence similarity, resulting in addition of 264 and 283 public SARS-CoV-2 predicted epitopes by allowing one and two amino acid mismatches, respectively. Moreover, to assess the risk of self-reactivity and immunopathology, we compared SARS-CoV-2 predicted epitopes with human proteome sequences and detected 10 predicted epitopes that are single amino acid variant from their counterparts in the human proteome. Lastly, to explore the potential for development of vaccines against coronavirus and influenza virus, we compared our list of predicted epitopes from coronaviruses with epitopes from influenza virus deposited in Immune Epitope Database and Analysis Resource (IEDB) and detected only a limited number of epitopes with a modest sequence similarity that are shared across multiple coronavirus strains and influenza.In this study, we investigated the level of T cell antigen cross-reactivity across the seven alpha and betacoronavirus strains, evaluated the risk of self-reactivity from SARS-CoV-2 predicted epitopes, and identified targets for vaccine developments against coronavirus and influenza virus. We first predicted the potential of peptides to be presented by 10 prevalent HLA alleles and eliciting CD8+ T cell responses, and generated a comprehensive The proteome sequences of coronavirus strains were obtained from NCBI. The reference numbers for these sequences are NC_002645.1 (229E), NC_006577.2 (HKU1), NC_019843.3 (MERS-CoV), NC_005831.2 (NL63), NC_006213.1 (OC43), NC_004718.3 (SARS-CoV) and NC_045512.2 (SARS-CoV-2).For multiple sequence alignment and phylogenetic tree generation, the open reading frames were grouped by their encoded proteins—spike protein (S), envelope protein (E), membrane protein (M), nucleocapsid protein (N), replicase polyprotein (Orf1ab), and other encoded regions (Other)—prior to analysis. The multiple sequence alignment was conducted using “msa” function from R msa v1.4.3 package and visualized by “msaPrettyPrint” function. The phylogenetic tree was produced by plotting “identity” distance generated by “dist.alignment” function from R seqinr v3.6.1 package.Each encoded proteins were fragmented into 9-mer peptides by scanning the proteome with a window of nine amino acids and step length of one amino acid. For strains containing two open reading frames annotated with the same functional protein e.g. 229E, MERS-CoV, SARS-CoV, and SARS-CoV-2 having two open reading frames annotated for Orf1ab, 9-mer peptides were generated from both encoded proteins and unique set of peptides were selected for subsequent analysis.The antigen presentation of MHC was predicted using NetMHCpan v4.0 , a modelhttps://github.com/masato-ogishi/Repitope.git).The immunogenicity potential was predicted by R package Repitope for peptides that passed NetMHCpan filtering i.e. those predicted to bind at least one HLA allele. The Repitope utilizes amino acid descriptors and TCR-peptide contact potential profiling (CPP)-based features to label immunogenicity. The Repitope package was retrieved from GitHub repository . Based on sequence alignments . Generally, there was a relatively high number of peptides predicted to bind HLA-C alleles while HLA-B alleles had the lowest number of predicted binders . These include HLA-A*0101, 0201, 0301, 2402, HLA-B*0702, 4001, 0801, and HLA-C*0702, 0401, 0701 . PrevaleRepitope  and identified in total 4,894 out of 16,096 (~30%) unique predicted binders to be immunogenic (subsequently referred to as predicted epitopes), and the proportion of such predicted epitopes was comparable across different strains . We retrieved all 7,869 linear epitopes in IEDB (as of 05-05-2020), presented on MHC-I, reported positive in T cell assays and have human as the host organism then compared with the list of 4,894 unique predicted epitopes from SARS-CoV-2.For a validation of predicted epitopes, we have compared the list of predicted epitopes with epitopes characterized by From the 7,869 IEBD immunogenic peptides, there were 34 unique peptides derived from coronavirusstrains, one from OC43, two from 229E, and the remaining from SARS-CoV. We identified 25 unique9-mer predicted epitopes from four coronavirus strains, 229E, NL63, SARS-CoV, and SARS-CoV-2 having matching pattern with 24 IEDB peptides  SARS-derived immunogenic peptides in IEDB matched with our 23 SARS-derived 9-mer predicted epitopes  SARS-CoV-2 predicted epitopes shared with four common coronaviruses with a high sequence similarity  deposited in IEDB.Due to a relative sequence dissimilarity between influenza virus and coronavirus, there were no peptides with identical match between two strains and all peptides were distinct by at least three amino acids  and reassortment among subtypes of virus (antigenic shift), there have been multiple attempts to develop generic vaccines protective against all influenza viruses to reduce the severity of infection and spread of disease , 28. Aloin silico profile of predicted epitopes shared across coronaviruses, we evaluated the potential risk of self-reactivity imposed by high sequence similarity with the human proteome. Considering the reports of lung, heart, liver, intestine, genital and kidney failures by autoimmune disorders in COVID-19 patients  increase in protein expressions after 20 h compared to 2 h post infection, with nucleocapsid and membrane proteins having the highest and the replicase polyprotein 1ab having the lowest fold change, respectively . TherefoComparing predicted epitopes with those characterized and deposited in IEDB, we could detect 64% of coronavirus IEDB epitopes from the prediction. In regards to the accuracy of predictive models utilized in this study, the algorithms to predict MHC presentation has matured significantly in the last decade by training with extensive datasets, especially for the most common HLA types. On the other hand, it is worth noting that predicting immunogenicity is challenging and not a fully solved problem. For example, these algorithms do not take into account other factors influencing the immunogenicity of a given epitope, such as abundance, expression pattern, and localization. Therefore, although the best performing models have been used for classifying immunogenicity, the predictions are suboptimal and should be taken with caution.γ secretion in the PBMC from HLA-A2 SARS-recovered donors ex vivo . While the presence of multiple 9-mers matching with the same IEDB epitope is not surprising, exclusion of some matches by the MHC binding and immunogenicity filtering may facilitate determining the regions of peptide that are most likely to elicit the T cell response.In this study, to validate our In a separate study, Nelde et al.  predicteNotably, this study further demonstrated presence of T cell responses in unexposed individuals to 31% of their validated HLA class I epitopes, which hints to presence of CD8+ T cell cross-reactivity with common coronaviruses. In agreement with our prediction, Orf1ab was the protein most recognized by the class I epitopes they found in healthy donors, while membrane (M) and nucleocapsid (N) proteins were not recognized. This points to the potential cross-reactivity with other coronavirus strains and suggests the significance of conceptualizing the map of cross-reactive potential among coronavirus strains.We believe that our comprehensive profile of private and public predicted epitopes across coronaviruses will assists biologists with targeted function validation, and facilitate design of vaccines capable of protecting against multiple prevalent virus strains. Further validations of the predicted profile would help estimate the extent of cross-protection and pave the way for better understanding of heterogeneity in the susceptibility and severity to the disease.All datasets presented in this study are included in the article/HK and GN conceived and designed the study. CL conceived and designed computational approaches, conducted data analysis, and generated figures and tables. PB conducted immunogenicity prediction. IW conducted MHC presentation prediction. CL MP, GO, AS, HK, and GN wrote the manuscript, and IW and PB contributed to methods. AS and HK supervised the project. All authors contributed to the article and approved the submitted version.GO has served on advisory boards or holds consultancies or equity with Eli Lilly, Novartis, Janssen, Sanofi, Orbit Discovery, and UCB Pharma, and has undertaken clinical trials with Atopix, Regeneron/Sanofi, and Roche.The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


In multivariable Cox regression analyses, a high CaScore was associated with overall graft failure (p = 0.004) and death with a functioning graft (p = 0.002), but not with death-censored graft failure and graft function decline. This study demonstrated that pre-transplant aorto-iliac calcification is associated with one-year eGFR in univariate, but not in multivariable linear regression analyses. Moreover, this study underlines that transplantation in patients with a high CaScore does not result in earlier transplant function decline or worse death censored graft survival, although ongoing efforts for the prevention of death with a functioning graft remain essential.While the association of vascular calcification with inferior patient outcomes in kidney transplant recipients is well-established, the association with graft outcomes has received less attention. With this dual-centre cohort study, we aimed to determine the clinical impact of recipient pre-transplant aorto-iliac calcification, measured on non-contrast enhanced computed tomography (CT)-imaging within three years prior to transplantation (2005–2018). We included 547 patients  years), with a median follow-up of 3.1 (1.4–5.2) years after transplantation. The aorto-iliac calcification score (CaScore) was inversely associated with one-year estimated-glomerular filtration rate (eGFR) in univariate linear regression analysis (standard β −3.3 (95% CI −5.1 to −1.5,  Atherosclerosis is an important pathophysiological process responsible for an increased disease burden in kidney transplant candidates and recipients, and is associated with decreased overall survival and increased cardiovascular morbidity and mortality ,2,3. CurKidney graft outcomes can be divided in short-term outcomes, such as early failure or one-year graft function, and long-term outcomes, such as (death-censored) graft failure and graft function decline . Death-cA recent meta-analysis on pre-transplant aorto-iliac calcification found an association with decreased overall survival and overall graft failure. Statistical significance was lacking for the association with death-censored graft failure and graft function. This lack of statistical power with regard to kidney graft outcomes can be attributed to the limited number of studies available, the various methods used for identification of calcification and the small sample sizes of the existing publications . Hence, We previously reported that the aorto-iliac calcification score (CaScore), the adjusted Agatston score, was independently associated with patient outcomes, e.g., risk of early (cardiovascular) death and cardiovascular events . In the We performed a dual-centre cohort study in The Netherlands  and Erasmus University Medical Center (Erasmus MC)), in adult kidney transplant recipients. All patients who underwent screening prior to transplantation by means of a non-contrast enhanced CT between January 2005 and December 2018 were included. CT procedures were performed according to the pre-transplant screening protocol in both transplant centres; selecting patients based on age (>50 years); comorbidities  and dialysis vintage (>2 years). Exclusion criteria were age <18 years at the time of transplantation and an interval of more than three years between CT and transplantation. In the inclusion period, both transplant centres participated in the Eurotranplant Senior Program (ESP), a Eurotransplant allocation scheme matching deceased donors above 65 years-of-age to recipients in a similar age range [The applied method for quantifying aorto-iliac calcification, as the Agatston score adjusted for the aorto-iliac trajectory (aorto-iliac CaScore) for non-enhanced CT images, has been described previously . In shor2), smoking status , hypercholesterolemia , total cholesterol level (mmol/L), systolic blood pressure (mmHg), use of antihypertensive medication, dialysis status , dialysis vintage (months), and number of previous kidney transplantations. The transplant variables included the type of donation , or donation after brain death (DBD)), donor gender and age, number of total human leukocyte antigen (HLA)-mismatches, warm ischemia time (minutes), and cold ischemia time (minute). Follow-up variables included eGFR at six-months and one-year, calculated with the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation (mL/min per 1.73 m2), laboratory values at one-year after transplantation , and urinary protein excretion), and primary cytomegalovirus (CMV) infection [Details of this cohort, including baseline cardiovascular risk factors, with history and follow-up of cardiovascular disease, have been published previously . The recnfection . Short-tnfection ,20.p-value <0.05 was considered significant.For baseline characteristics, patients were stratified into two groups based on the aorto-iliac CaScore, being a low CaScore (0–5600 HU) and a high CaScore (>5600 HU). These cut-off values are based on the initial aorto-iliac CaScore analyses, with the high CaScore derived from the highest tertile of CaScores . Data we(Death-censored) graft failure-free and graft function decline-free were calculated using Kaplan-Meier survival curves. The low and high aorto-iliac CaScores were compared for living and deceased-donor kidney transplant recipients with Log-rank testing. Cox proportional hazards regression analysis, for living and deceased-donor kidney transplant recipients, was used to establish the association of the high aorto-iliac CaScore with (death-censored) graft failure, death with a functioning graft and graft function decline. Linear regression was used to establish the association of the aorto-iliac CaScore (continuous) with one-year eGFR. Models were built with a priori selected covariables. In Model 1, we adjusted for transplant centre (UMCG or Erasmus MC) and time between CT and transplantation. In model 2, we adjusted for model 1 plus donor gender, donor type , cold ischemia time, number of HLA mismatches, recipient gender, diabetes mellitus, smoking, dialysis vintage, number of previous transplantations, statin use pre-transplantation. In model 3, we adjusted for model 2 plus recipient age and in model 4 for model 3 plus donor age. For linear regression, laboratory results at one year after transplantation  and an episode of acute rejection were included as additional variables in model 2. Analysis of interaction terms for the association of the aorto-iliac CaScore (continuous) with one-year eGFR by the covariables recipient age, gender, diabetes, donor age, gender, and donor type, was performed. To quantify the magnitude of confounding by donor age on the association between the aorto-iliac CaScore and one-year eGFR, we used the regression methodology as proposed by Janes et al. . In addiStatistical analyses were performed with R: A Language and Environment for Statistical Computing, version 1.0.153 for Mac , using the software R-Packages “MASS”, “MICE” “mediation”, “survival”, “ggplot2”, and “survminer”.n = 462) of the CaScore groups differed for eGFR at six-months and one-year, calcium, phosphate, glucose, PTH, and urinary protein excretion (n = 9 (4.9) vs. n = 5 (1.4), p = 0.027), but no differences in delayed graft function or acute rejections  years). The mean time from CT to transplantation was 0.91 (0.72) years and the median follow-up was 3.1  years after transplantation. xcretion . For shojections . In ninen = 462) was 51 (21), with 48 (21) in the high and 53 (20) in the low CaScore group. The CaScore (continuous) was inversely associated with one-year eGFR, with a standard β −3.5  when adjusting for transplant centre and time CT to transplantation (model 1). When adjusting for patient, transplantation and follow-up covariables, with the exception of donor age, this inverse association remained significant , and model 3 (including recipient age), standard β −2.7 ). This association lost statistical significance after adjustment for donor age ) (p < 0.0001), but not for the covariables recipient age, gender, diabetes, donor gender, and donor type (p > 0.05). When quantifying the magnitude of confounding, the direct effect of the aorto-iliac CaScore on one-year eGFR was −3.3 (95% CI −5.1 to −1.5) and the effect on the confounder (donor age) was 5.0 (95% CI 3.8 to 6.2). The direct effect of the confounder on one-year eGFR was −0.6 (95% CI −0.7 to −0.5), with a bootstrapped indirect effect of −2.9 (95% CI −4.0 to −1.9). Donor age was identified as a confounder of the association between the aorto-iliac CaScore and one-year eGFR, resulting in an 87.2% of the association (Mean eGFR at one-year after transplantation ( 0.077)) . There wociation . In Tablp < 0.001) )  events of death-censored graft failure occurred, with 23 (12.6%) in the high and 41 (11.2%) in the low CaScore group . Kaplan–< 0.001) . Kaplan– 0.711)) . No diff 0.711)) .p < 0.0001) (p < 0.0001) (p = 0.006)) (p = 0.048) and for living-donation   events of overall graft failure occurred, with 54 (29.7%) in the high and 64 (17.5%) in the low CaScore group . Kaplan– 0.0001) . Kaplan– 0.0001) . In Cox  0.006)) . When st 0.0036) .p < 0.0001) (p < 0.0001) (p = 0.224))   events of death with a functioning graft occurred, with 31 (17.0%) in the high and 23 (6.3%) in the low CaScore group . Kaplan– 0.0001) . Kaplan- 0.0001) . In Cox  0.224)) . When stctively) .p < 0.0001) (p = 0.014) (p = 0.223)  events of graft function decline occurred, with 51 (14.0%) in the high and 30 (16.5%) in the low CaScore group . Kaplan– 0.0001) . Kaplan–= 0.014) . In Cox = 0.223) . No diff= 0.223) .In this dual centre cohort study, we identified a non-causal inverse association between pre-transplant aorto-iliac calcification and one-year eGFR in univariate analysis. This association was not significant after adjustment for patient-specific, transplantation-related, and follow-up covariables, including donor and recipient age. The increased donor age in older recipients, due to the allocation of older donor kidneys to older recipients in the Eurotransplant Senior Program, was a significant confounder. For long-term graft outcomes, no association could be established between a high aorto-iliac CaScore and an increased risk of death-censored graft failure or graft function decline, in both living and deceased donor kidney transplantation. Whereas the identified association between a high aorto-iliac CaScore and graft failure (death-censored) remained significant after adjustment for various covariables, including donor and recipient age. Also, the high aorto-iliac CaScore was associated with death with a functioning graft.n = 374), aorto-iliac stenosis on CT-imaging was not an independent risk factor for graft failure, but was associated with an increased risk of overall graft failure [n = 119) with lateral lumbar radiography-based calcification assessment, transplant recipients with and without calcifications differed with regard to graft failure-free survival [n = 100), no statistically significant difference in eGFR at one-year after transplantation was found for the quartiles of aorto-iliac calcification. Also, survival analysis for graft failure, with nine events after a median follow-up of 2.6 years, did not show differences between the quartiles [Our study has both similarities and important differences compared to four previously published studies on pre-transplant imaging-based visualization of calcification. In the largest study , differences in graft failure were observed for transplant recipients with and without intimal microcalcification of the iliac artery; however, these differences were not tested in multivariable analyses. For graft function decline, an independent association with the combination of intimal microcalcification with positive intimal fetuin-A was identified [n = 699), calciprotein particle maturation time (T50), a measure for blood calcification propensity, were shown to be associated with all-cause mortality, cardiovascular mortality, and graft failure [The outcomes of our study are in line with previous non-imaging studies on calcification. In a histology-based cohort study  laboratory values at one-year after transplantation were not available. Complying with general standards for dealing with missing data, a complete case analysis was found to be most reliable. In addition, linear regression was performed after multiple imputation, with imputations for serum PTH and proteinuria. Fifth, the interpretation of the implications of a statistically significant difference should always include a reflection of the clinical significance of the observed difference. In this study, a mean difference in eGFR of 5 mL/min per 1.73 m2 was found between a high and a low aorto-iliac CaScore (mean 48 (21) vs. 53 (20)). With a mean eGFR of 51 in the total cohort, this is a 10% difference in one-year graft function.Our study has a few limitations that need to be addressed. First, we retrospectively selected patients in which a CT procedure was already performed based on predefined criteria, making the study prone to selection bias and confounding. Although the presented findings are applicable to the majority of transplant recipients with comparable baseline characteristics (median age of 60 (51–68) years), they should not be extrapolated to relatively young patients with limited comorbidities. Second, the two transplant centres in this cohort study are part of Eurotransplant Senior Program ,38. ThisNonetheless, this study has its unique strengths, which justify attention. This is the first study evaluating the association between aorto-iliac calcification and one-year graft function in multivariable analysis, including patient, transplantation, and follow-up covariables. Second, the aorto-iliac CaScore is an easy-to-implement assessment, as the software is readily available in standard imaging analysis packages and pre-transplant CT-imaging is advised for high-risk transplant candidates ,40. ThirIn conclusion, this study demonstrated that pre-transplant aorto-iliac calcification is associated with one-year eGFR in univariate, but not in multivariable linear regression analyses. The increased donor age in older recipients is a significant confounder for this association. A high CaScore was not associated with death-censored graft failure and graft function decline, whereas independent associations with overall graft failure and death with a functioning graft were identified. These results underline that transplantation in patients with a high CaScore does not result in earlier transplant function decline or worse death censored graft survival, although ongoing efforts for the prevention of death with a functioning graft remain essential.


Dynamic EMG analysis showed improved phasic gastrocnemius activity (p < 0.0001). Equinus gait was improved with the reduction of peak plantar flexion (p < 0.0001), as well as an increase in peak dorsiflexion (p = 0.006) during walking was observed. Ankle joint quasi-stiffness  decreased significantly (p = 0.0017) after SDR. Moreover, even though the total time of the gait cycle did not change (p = 0.99), the time interval from maximum dorsiflexion to maximum plantar flexion  increased significantly (p = 0.0068) after SDR. In conclusion, the decreased ankle quasi-stiffness and the enhanced time interval in the gait cycle due to SDR indicate better motor control and joint stability. Our findings suggest that the long-term contracture formation occurring even after surgical interventions may be related to the stiffening of non-contractile structures.Selective dorsal rhizotomy (SDR) is a neurosurgical technique performed to reduce muscle spasticity and improve motor functions in children with cerebral palsy (CP). In long term, muscle contractures were observed even after SDR. To better understand what is contributing to contracture formation, it is necessary to assess the effects of SDR on joint stiffness. We hypothesized that ankle passive range of motion (ROM) increases and the quasi-stiffness of the ankle joint decreases after SDR in children with CP. This retrospective study included 10 children with diplegic CP (median age 6 years 2 months) who had undergone SDR and for whom gait analysis data were collected 3 months before (Pre-SDR) and 13 months after (Post-SDR) surgery. Additional to clinical measures, ankle quasi-stiffness (the slope of the ankle moment vs. ankle angle plot) was analyzed from gait data. Passive ankle ROM at 0° ( Selective dorsal rhizotomy (SDR) is a well-established neurosurgical technique performed to reduce muscle spasticity and improve motor functions in children with cerebral palsy (CP) –3. DurinPatients with CP develop muscle contractures that are thought to be multifactorial, including chronic muscle shortening  and decrThe present study was approved by the Institutional Review Board of Mayo Clinic, Rochester, MN. A retrospective medical record review was performed to identify children (18 years of age or younger) with a diagnosis of spastic diplegic CP who had undergone SDR. Children were included if they had received gait analysis evaluations within 18 months before (Pre-SDR) and 18 months after (Post-SDR) surgery. Children were excluded from the study if ankle kinetic or dynamic EMG data were missing.One to two-level laminectomy procedures were applied to patients. Electrical stimulation was used to identify sensory roots from L2-S1 levels on both the left and right sides. The sensory roots at these levels were then separated into rootlets, with the number of rootlets sectioned per level varying depending on the electrophysiological response. In general, a sensory nerve was separated into 3–10 rootlets with one-third to three-fourths of the rootlets sectioned per side at each level. Taking into account all the sensory rootlets stimulated from L2-S1 levels, ~40–60% of rootlets were sectioned.The Gross Motor Functional Classification System (GMFCS)  was usedA set of 3D markers was placed on the body of each subject, including the sacrum and bilaterally on the acromion processes, lateral epicondyle of the elbows, center of the dorsum of the wrists, anterior superior iliac spines (ASIS), lateral femoral condyles, lateral malleoli, mid-thigh, mid-shank, heels, and the spaces between the first and second metatarsal heads. The model used was the same as described by Kadaba et al.  except fFor gait analysis, a 10-camera motion capture system  was used. The 3D coordinates of the markers and force plate were used as inputs to a commercial software program , to calculate the joint kinematics and kinetics. The Visual3D program was used to define the joint center and segment coordinate systems from the 3D marker trajectories, as well as the subsequent rigid body kinematic/kinetic calculations. Embedded right-hand Cartesian coordinate systems were used in this model to describe the position and orientation of the rigid body segments of the lower extremity. A three-dimensional coordinate system was used for the foot with an anteriorly directed x-axis, y-axis pointing to the body's left side, and a superiorly directed z-axis. With these embedded coordinate systems, the joint angles were determined using the floating-axis or Euler angle convention. The ankle center was located by a vector directed medially from the lateral malleolus marker at one-half the distance of the measured ankle width. The segmental joint forces and moments were derived based on the calculated joint centers.A 16 channel surface EMG system  was used for data collection and synchronized with the kinematic data. EMG data were collected at 2,400 Hz from the rectus femoris, hamstrings, tibialis anterior, peroneus longus, and gastrocnemius muscles rectified and passed through a fourth-order low pass Butterworth filter with 6 Hz cutoff using custom Matlab  programming. Placement of EMG electrodes is confirmed by real-time visual observation of the EMG signal while muscle activation is achieved through resisted motion.Ankle passive ROM was recorded at two knee angles: (i) The patient was positioned supine with the knee fully extended (at 0°) and (ii) the patient was positioned prone with the knee at 90°. The foot was positioned in neutral inversion and eversion. The proximal arm of the goniometer was aligned with the lateral midline of the lower leg using the head of the fibula as a reference point. By aligning the distal arm of the goniometer parallel to the fifth metatarsal, the examiner changed the ankle angle slowly to measure max PF and max DF.To quantify the effects of SDR on the ankle angle used during walking, the maximum PF and DF angles were calculated from the average of three trials for each participant.The Visual3D computer software package along with a set of custom Matlab  programs were used for data reduction and database archival. Three trials were collected from each participant. The period of a gait cycle (stride) was defined from the initial contact of one lower extremity to the next initial contact of that same extremity. Kinematic measurements included the motion of the body and limb segments during representative walking strides. The kinetic analysis included the net forces and torques (moment) exerted on the body as a result of the combined effects of the ground reaction force, inertia, and muscle contraction. The joint moments were described using an internal moment convention. 3D kinematics was reported in degrees, and kinetics was normalized to body mass. By using raw time-series kinematics and kinetics, the ankle moment was calculated as a function of ankle PF-DF angles for each trial. During the first max PF in early stance to max DF in midstance ; the shank moving forward is controlled by the eccentric contraction of the gastrocnemius and soleus. Davis and DeLuca  describeMoreover, as there has been concerns raised regarding the precision of the term “dynamic stiffness” in this context, previous literature  introducr ≥ 0.80), the calculated slope was recorded as ankle joint quasi-stiffness. If high correlations were observed for more than one trial for each limb, the average of the slopes of trials was recorded as quasi-stiffness.The slope of the moment (Nm/kg) vs. ankle angle (degrees) at this specific time interval was determined using a linear regression model . The PeaEnhanced gastrocnemius muscle tone is defined as one of the major determinants of ankle joint stiffness. Therefore, the phasic character of the gastrocnemius muscle was scored by using dynamic EMG data with respect to the gait cycle: 0, Phasic; 1, Increased Phasic Activity; 2, Continuous Low-Level Activity; 3, Continuous with Phasic Pattern; 4, Continuous High-Level Activity . Scoringrs) was calculated between the absolute change in ankle passive ROM and quasi-stiffness due to SDR. Differences were considered significant if p < 0.05.For statistical analyses, SAS 9.4  software was used. To take into account that right and left limbs may provide correlated response data, generalized estimating equations were used separately to evaluate if significant changes occurred in (i) ankle joint passive ROM, (ii) dynamic EMG scores, (iii) max PF and DF angles during walking, (iv) joint quasi-stiffness, (v) the time interval from max DF to max PF, and (vi) total gait cycle time between Pre-SDR and Post-SDR. To test the correlation between the effects of SDR on passive and dynamic parameters, Spearman's rank correlation coefficient . Pre-SDR and post-SDR clinical tests and gait analyses were performed 3 months  before surgery and 13 months  after surgery, respectively. Two children were operated on unilaterally. The data collected from their non-operated legs were excluded from the analyses. Pre-SDR GMFCS levels ranged from I to III. GMFCS levels of half of the children improved post-SDR .p < 0.0001)    and max DF increased significantly (p < 0.0001) .After SDR, max PF during walking decreased significantly  after SDR surgery (p = 0.99), the time interval from max DF to max PF  significantly increased (p = 0.0068) after SDR surgery  decreased significantly ( surgery . Moreove surgery .rs = 0.36 for ankle ROM at 0° knee angle vs. stiffness (p = 0.30) and rs = 0.24 for ankle ROM at 90° knee angle vs. stiffness (p = 0.41)].No significant correlation was found between the changes in ankle passive ROM and the changes in quasi-stiffness [The present study showed increased passive ankle joint ROM and decreased quasi-stiffness for children with CP after SDR. These results support both of our hypotheses. Furthermore, we found that the time interval from max DF to max PF during gait was elevated substantially by SDR. Numerous studies , 6, 11 hIncreased passive ROM of ankle joint both at knee flexion and extension indicates improvement in passive mechanical characteristics of all plantar flexor muscles. Additionally, gastrocnemius EMG revealed an improved phasic activity after SDR. This also contributed to a reduction in passive joint stiffness. As the children with CP underwent physical therapy (per our program's post-SDR therapy protocol), the improvement in passive ROM may be more associated with the physical therapy intervention, than SDR alone. A previous study  showed tWe found that the dynamic gait characteristics of the ankle joint improve after SDR. Specifically, a significant increase in max DF and reduction of max PF indicate a substantial shift of ankle angles used during walking. The shift in ankle angles suggests a reduction in equinus gait after SDR in children with CP, which is consistent with previous gait analysis data , 29. ThuOn the other hand, the changes in passive ankle ROM do not correlate with the joint quasi-stiffness. This could be due to the fact that the passive behavior of joints comprises not only the passive muscle characteristics but also the amount and the stiffness of non-muscular structures such as fascial connective tissues, aponeurosis, ligaments, and nerves , 31. Thein vivo is unrelated to the stretch reflex for spastic vastus lateralis of spinal cord injured patients by using EMG and dynamometer data collected during knee flexion and extension. From the biopsied muscle samples, they showed fiber type alterations and higher passive tension at the cell level due to spasticity as well  (#16-010165).FA, JB, and KK contributed to the conception and design of the study as well as the interpretation of the results. FA performed data analysis and manuscript writing. KK contributed to writing, editing, and critical appraisal of the manuscript. All authors approved the final submitted manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


R2 = 0.96) with the melting enthalpies of retrograded starches, which, in turn, were negatively correlated with the extent (R2 = 0.92) and rate (R2 = 0.79) of in vitro digestion. It was concluded that AMM has the potential to be used to deliver novel starch functionalities and enhance its nutritional properties.The fine molecular structure of starch governs its functionality and digestibility, and enzymatic approaches can be utilized to tailor its properties. The aim of this study was to investigate the in situ modification of starch by amylomaltase (AMM) from Thermus thermophilus in model starch systems subjected to hydrothermal treatments under standardized conditions and the relationship between molecular structure, rheological properties and in vitro digestibility. When low dosages of AMM were added to a wheat starch suspension prior to submitting it to a temperature-time profile in a Rapid Visco Analyzer, the increased peak viscosity observed was attributed to partial depolymerization of amylose, which facilitated starch swelling and viscosity development. At higher dosages, the effect was smaller. The low cold paste viscosity as a result of the activity of AMM reflected substantial amylose depolymerization. At the same time, amylopectin chains were substantially elongated. The longer amylopectin chains were positively correlated ( Starch is the most abundant glycemic carbohydrate in the human diet. In the gastrointestinal tract, it is mainly digested by salivary and pancreatic α-amylase and intestinal mucosal α-glucosidases. Pancreatic α-amylase internally hydrolyzes α-1,4-glycosidic bonds, thereby releasing short-chain hydrolysis products, namely maltose, maltotriose and α-limit dextrins. The latter contain undigested α-1,6-glycosidic bonds . UndoubtStarch consists of two polymers, i.e., amylose (AM) and amylopectin (AP), which differ in susceptibility to digestive α-amylases.AM is an essentially linear polymer. It consists of glucose units predominantly linked by α-1,4-glycosidic bonds. AP is a highly branched polymer with approximately 5% of its glycosidic bonds being of the α-1,6 type , and in Likewise, the molecular structure of AP, and structural aspects such as average chain length and branching density, have a huge impact on starch digestion. The parabolic relationship between (i) the ratio of short to long AP chains of cooked and retrograded maize starch, and (ii) the content of slowly digestible starch, shows that starches dominantly containing either short or long AP chains contain high levels of slowly digestible starch . High coThermus thermophilus is active at temperatures exceeding those at which starch gelatinizes  holds potential in this respect. This 4-α-glucanotransferase catalyzes the transfer of a segment of α-1,4-D-glucan to a new 4-position in an acceptor carbohydrate via a disproportionation reaction . When AMis 72 °C ) and is is 72 °C . In addiis 72 °C  that leais 72 °C . Howeveris 72 °C , and theis 72 °C , it is eAcidothermus cellulolyticus 11B led to a decrease in rapidly digestible starch content from 80% to 60%, and an increase of slowly digestible starch content from 12% up to 20% , porcine pancreatin  and isoamylase from Pseudomonas sp (08124) preparations purchased from Sigma-Aldrich , unless otherwise specified. Wheat starch was obtained from Tereos Syral . A liquid preparation of AMM from w/v maltotriose and 0.1% w/v maltotetraose was incubated with 0.1 mL properly diluted enzyme preparation in the same buffer for up to 30 min at 70 °C. Samples were then boiled for 30 min to inactivate the enzyme. For each time point , a different tube was prepared. The released glucose was measured using a colorimetric assay . One enzyme unit (U) of AMM enzyme activity is defined as the amount of enzyme that under the assay conditions produces 1 μmol of glucose per min at pH 6.5 and 70 °C.The AMM activity was assayed in triplicate according to van der Maarel et al. . Thus, 0w/w dry matter (dm)] was suspended in 100 mM sodium maleate buffer (pH 6.0) containing 5.0 mM CaCl2 and AMM  to obtain 25.0 g final sample weight. The starch suspension was equilibrated at 40 °C for 1 min and the temperature was then increased to 95 °C in 12 min, held constant at 95 °C for 5 min and decreased to 50 °C in 7 min before final isothermal holding at 50 °C for 30 min. The slurries were stirred at 160 rpm throughout analysis. Peak viscosity is the maximum viscosity during heating and the cold paste viscosity that at the end of the rapid visco analysis (RVA) profile. Breakdown viscosity is the difference between peak and minimum viscosity during the 95 °C holding phase. The latter is the hot paste viscosity and the difference between cold and hot paste viscosity is the setback. After the RVA run, the starch slurries were stored at 5 °C for 24 h, frozen with liquid nitrogen and freeze-dried. The dried material was gently ground with mortar and pestle, sieved (aperture: 250 μm) and stored in closed containers at room temperature until further analysis. In addition, for another set of samples used to study the molecular structure of AM and AP, the RVA profile was stopped when the peak viscosity was reached and the starch slurries were immediately frozen with liquid nitrogen and freeze-dried.The rheological properties of starch suspensions during in situ starch modification by AMM were studied in triplicate with a dose-response approach with a Rapid Visco Analyzer Super 4  based on Derde et al. . Wheat sThe fine molecular structure of AM and AP was studied as in Dries et al.  by using3 (DP 8), 6.2 × 103 (DP 38), 10.0 × 103 (DP 61), 23.0 ×103 (DP 141), 48.8 × 103 (DP 301), 113.0 × 103 (DP 697), 200 × 103 (DP 1234), 348 × 103 (DP 2148) and 805 × 103 (DP 4969). The logarithm of the molecular weights of the standards versus their corresponding elution volumes were plotted and fitted with a third order polynomial equation (R2 = 0.999). The first (DP < 100) and second  peak of the weight distributions corresponded to debranched AP and debranched AM, respectively [The chain length distributions of debranched AM and AP samples were studied with HPSEC as described by Dries et al. . Debrancectively . Furtherectively . w/w) sample dm:water] was added. Pans were hermetically sealed and left at room temperature for 45 min. They were then further equilibrated at 0 °C before being heated from 0 to 120 °C at 4 °C/min. The peak temperatures [Tp (°C)] and enthalpies [ΔH (J/g starch dm)] of the melting of AP crystals formed as a result of retrogradation were determined with the TA Universal Analysis software.The extent of retrogradation of starch slurries first prepared in the RVA and then stored for 24 h at 5 °C was studied by differential scanning calorimetry (DSC) with a Q2000 DSC  instrument. The samples were first frozen with liquid nitrogen and freeze-dried. Freeze-dried starch samples were accurately weighed (2.0–3.0 mg) in aluminum pans  and water [1:5 (2 (200 mM) and MgCl2 (0.49 mM) was added. The digestion mixture was incubated for 90 min at 37 °C under magnetic stirring (200 rpm). At predetermined time points, 0.1 mL aliquots were transferred into Eppendorf tubes containing 0.2 mL 300 mM sodium carbonate to terminate starch digestion. Following shaking and centrifugation , the reducing sugar contents in the supernatants were quantified by the method of Moretti and Thorson [The in vitro starch digestion assay was based on Korompokis et al. . An aliq Thorson  using tht: the percentage of starch digested (%), t: incubation time (min), C∞: percentage of starch digested (%) at infinite time and k: rate constant (min−1). For fitting purposes, the baseline value at t = 0 min was subtracted from the subsequent values at t time. The fitting was performed with the statistical software JMP Pro 14.0 . The different digestion curves were fitted to a first-order equation as described by Goñi et al.  in EquatStatistical significance of differences between mean values was assessed by one-way analysis of variance (ANOVA) followed by post hoc Tukey’s test. The confidence level was 95% and the statistical analysis was performed with JMP Pro 14.0 . Different dosages of AMM influenced the viscosity of starch slurries differently. Three AMM dosage categories were considered: low (0.45 U/g starch dm), intermediate (1.8–9 U/g starch dm) and high (18–45 U/g starch dm) . The low dosage A did notAnalysis of the branches of control and AMM treated starches for unmodified native wheat starch revealed two characteristic distributions in the HPSEC profiles . The firWhile the low AMM concentration only had a minor effect on the AP chain length distribution, it did result in considerable shortening of the long AM chains (DP range decreased to a range of 100–2500) A. At intp readings drastically increased with AMM dosages. While Tp was 50 °C for the control starch, Tp readings were higher  for AMM treated samples.Control starch and starch modified with the low AMM dosage resulted in low ΔH (about 0.4 to 0.8 J/g starch dm) readings . At inteAMM action during the RVA run had a profound impact on AM and AP molecular structure. The AM population was gradually eliminated and the AP population increasingly had longer chains . AMM canStarch structure was not only studied for samples withdrawn at the end of the RVA run but also when the viscosity reached its peak. Already at this point in the RVA run were the HPSEC profiles similar  to thoseIn this study, a model starch system was used to study the potential of AMM to modify starch in situ during hydrothermal processing. To some extent, doing so mimicked food processing conditions wherein enzymes are included in the initial ingredient mix and act when starch gelatinizes during heating. At the same time, while such conditions do not fully represent the conditions for producing a whole range of various starch-containing food products, similar model systems have been effectively utilized in studies of the enzyme functionality in bread systems ,29,30.The viscosity of the starch slurries was clearly affected by in situ action of AMM in a dose-dependent way. The low enzyme dosage led to higher hot paste, lower breakdown and setback and moderately higher cold paste viscosity A. PartiaFurther modifications at intermediate enzyme dosages led to improved swelling and viscosity development but also to weaker gels B. To theWhen high enzyme dosages were used, there was a minor increase in the peak viscosity and a continuous decrease in hot paste, cold paste and setback viscosities. Peak viscosities were reached earlier as a result of the enzyme action C. The hiThe decreased peak viscosities after the use of high AMM dosages compared to those resulting from the use of intermediate AMM dosages may indicate more extensive AP degradation as a whole. Indeed, amylolytic activity has been reported at high AMM dosages . Once AMOverall, the impact of different AMM dosages on the viscosity of starch slurries could be directly related to the extent of AM structure modification. Low dosages shortened AM chains to some extent and thus made them more mobile. This positively impacted hot and cold paste viscosities due to enhanced AM—AM interactions. At intermediate levels, the effect on AM depletion was more dominant. This led to more drastic changes in rheological behavior and highlighted the role of AM in starch swelling and viscosity development. High levels of AMM did not induce further changes in the molecular structure of starch polymers and consequently, only had limited additional impact. However, they did result in an increased rate of swelling and viscosity development. p readings were detected due, reasonably, to (hardly) unaltered AP structures. The use of intermediate dosages resulted in both drastically increased ΔH and Tp values. Starch with elongated AP chains has a higher propensity and/or potential to retrograde. As also reported by Hansen et al. [R2 = 0.96) correlated to the ΔH values  and k (R2 = 0.93), respectively (∞ (At intermediate concentrations, both the extent and rate of digestion significantly declined . That coectively . Higher ∞ R2 = 0.1 and k (ively (∞ A and ΔH ively (∞ B showed Upon digestion, pancreatic α-amylase randomly cleaves α-1,4-glycosidic bonds in an endoactivity pattern and releases starch chains of various DP which are further completely digested to release maltose, maltotriose and α-limit dextrins ,43. WittLastly, at high dosage there was no additional effect on starch digestibility compared to that resulting from the intermediate dosage. This was expected since there were no major additional modifications on AP structure governing its ability to form crystalline structures and, thus, to lower starch digestibility. The slight amylolytic activity had no effect on starch digestibility. It is clear from the above that the extent of AP chain length modification by AMM governs starch digestibility. At low AMM dosage, there was no significant impact on AP structure, and consequently, only a limited impact on starch digestion. At intermediate dosages, elongated AP chains were more abundant and formed more and/or more easily crystalline structures upon cold storage at 5 °C for 24 h, and starch digestibility was evidently decreased. Since the high dosages had no additional effect on AP structure, no additional impact on starch digestion was observed. From a nutritional point of view, then, adequately chosen AMM dosages can sufficiently elongate AP chains and lower starch digestibility.In this work, the impact of various AMM dosages on the molecular structure of starch and the subsequent effect on starch rheological properties and digestibility was studied for the first time. As schematically presented in , as a reIn conclusion, the use of AMM induces novel starch functionality and the considerable decrease in starch digestibility imparts AMM with a great application potential. The holistic approach in this study gives a basic understanding of how different AMM dosages can variably influence the physicochemical properties and digestibility of starch. Insights from this fundamental study can be of great use in the context of the application of AMM for producing starch-containing foods. AMM can thus be utilized in the production of a range of starchy food products as a processing agent to manipulate viscosity when deemed useful. Furthermore, AMM can be used in the production of starchy foods with lower starch digestibility and sustained glucose release that provide the consumer with healthier options. The use of starch-modifying enzymes in foods with the goal of lowering their glycemic potential is not commonly practiced in food industry. Hence, knowledge on enzymes such as AMM can contribute to novel approaches to improve food nutritional properties without compromising quality.


Dear colleagues, dear editors, dear reviewers, dear authors,After more than 20 years of work with the International Journal of Legal Medicine, it is time for me to relinquish my role as Editor-in-Chief. Therefore, I would like to take this opportunity to thank you all for your trusting cooperation over the previous years. Without the commitment of all of you it would not have been possible to develop our journal as successfully as it is. It was always a pleasure for me to interact with authors and reviewers and to have the opportunity to accompany a high number of excellent manuscripts on their way to publication. I would like to thank Prof. Pfeiffer and Prof. Schmeling as well as Mrs. Beckhoff for years of trusting and friendly collaboration. Furthermore, I wish to thank all the friends from the presidium of the International Academy of Legal Medicine, in particularly the past president, Prof. Davide Ferrara, and the actual president, Prof. Angel Carracedo. I have really enjoyed working with you. Finally yet importantly, I would like to thank the Springer representatives who are/were responsible for the Journal from the publisher site. Dr. Pillmann an now Mrs. Ullmann supported the editors in all important organizational questions and had/has a significant influence on the expansion of the content and scope of the journal.At the beginning of next year, Prof. Schmeling will follow me as Editor-in-Chief, and my colleague and friend Prof. Tony Fracasso will follow me as associated editor who is responsible for morphology and clinical forensic medicine. As you all know, both are extraordinary scientists, one is a specialist in anthropology and the other in forensic morphology. I am sure that Prof. Fracasso, the new member of the team will support the editorial team by excellent work. I wish him all the best for this new task.Best wishes to all of you and for the journal.Thomas Bajanowski


The dominantly inherited spinocerebellar ataxias (SCAs) are a large class of neurodegenerative diseases. Transcranial magnetic stimulation has been used to evaluate the function of the pyramidal tract, and central motor conduction time (CMCT) is one index used to detect pyramidal tract dysfunction. We conducted a comprehensive search of PubMed, Embase and Web of Science. Eight eligible studies were included in the meta-analysis. For upper limb CMCT, the mean difference ) between the combined SCA group and the control group was 2.24 [1.76-2.72], while the mean differences (95% CIs) between the subtypes and the control group were as follows: 4.43 [3.58-5.28] for SCA1, 0.25  for SCA2, 1.04  for SCA3 and 0.49  for SCA6. Additionally, SCA1 significantly differed from SCA2 and SCA3 in terms of CMCT . We also compared lower limb CMCT between the SCA2 and control groups. The mean difference (95% CI) was 6.58 [4.49-8.67], which was clearly statistically significant. The differences in CMCT values among different subtypes suggests diverse pathological mechanisms. In general, CMCT is a promising objective index to judge the severity of disease deserving further investigation. The dominantly inherited spinocerebellar ataxias (SCAs) are a large class of neurodegenerative diseases . More thTranscranial magnetic stimulation (TMS) is a relatively safe and noninvasive means of electrically stimulating the brain by electromagnetic induction. This technique is currently being used in the diagnosis and treatment of many diseases. Because of its convenience and noninvasiveness, TMS has a high rate of patient compliance , 9. TMS Studies that address this question are scarce. In some previous studies, TMS was used in patients with SCAs, and CMCT showed different degrees of prolongation in patients compared with the control group . HoweverBy using keywords in the PubMed, Embase and Web of Science databases, we obtained 648 records after removing duplicate records. After the preliminary reading of the title and abstract, 16 articles that fit the topic were read in full-text form. Based on the exclusion criteria, 8 articles were eliminated , but after we excluded the study by T Yokota (1998), the heterogeneity of three subtype comparisons became extremely low (I2 = 0%). We observed that the sample sizes for SCA1, SCA2 and SCA3 were all 18 , and significant differences were observed , and the difference was statistically significant .Because there were very few data on lower limb CMCT, we analyzed only the data from the SCA2 and control groups. The results are displayed in This study is the first meta-analysis of CMCT in patients with SCAs. Compared with the control group, the SCA showed a trend toward prolonged CMCT of the upper limb, although there was some heterogeneity. To analyze and reduce the heterogeneity, we organized the analysis into groups according to SCA subtype. Among the various subtypes, the difference was most obvious for SCA1. In the SCA2, SCA3, and SCA6 groups, the CMCT of the upper limbs was not significantly different from that of the control group. The comparison among subtypes showed that SCA1 was significantly different from SCA2 and SCA3. For the lower limbs, patients with SCA2 showed significantly prolonged CMCT compared to the control group.Sensitivity analysis revealed that the three articles on SCA1 had good homogeneity, while the four articles on SCA3 had high heterogeneity. The reasons for the high heterogeneity in SCA3 might be as follows. First, the sample size was quite small. Second, The reasons for the heterogeneity introduced by those two studies are as follows. First, there were differences in the characteristics of the study populations and the control groups. For example, in the study of SCA2 by Ketan Jhunjhunwala 2013) [13 [20], SCA1, 2 and 3 are autosomal dominant disorders characterized by cerebellar ataxia; these disorders manifest as oculomotor nerve abnormalities, pyramidal or extrapyramidal features and peripheral neuropathy. CMCT is a quantitative neurophysiological measurement method for objectively evaluating corticospinal tract conduction. This measurement reflects the conduction between the primary motor cortex and spinal cord. There may be several mechanisms of CMCT prolongation. First, axons or synapses in the pyramidal tract may gradually decrease in number over the course of the disease. In addition, axons of the pyramidal tract may become demyelinated or atrophied. Moreover, decreased excitability of the α-motor neurons may prolong the time it takes for the membrane potential of those neurons to reach the threshold for an action potential . PreviouIn general, compared with the control group, CMCT of SCA1 and SCA2 types was significantly prolonged, while there was no significant difference in SCA3 or SCA6 types. Our meta-analysis provides a new direction for future studies on the pathophysiology of SCAs. First, the difference in CMCT subtypes could reflect the involvement of the motor cortex and conduction pathway, thus providing clinical evidence for the pathology of the disease. Second, there were obvious differences in CMCT among the SCA1, SCA2, SCA3 and control groups, suggesting that CMCT could be used as an auxiliary means of disease classification. Additionally, as a noninvasive examination, it could be used as a potential follow-up evaluation index alongside a clinical scale in cohort studies to estimate the progression of the disease and the effect of therapy.We have several suggestions for future research. Above all, it would be best to expand the sample size and ensure a lack of confounders between patients and controls. Furthermore, TMS methods and parameters should be standardized to facilitate the sorting and analysis of the results. Both upper limb and lower limb CMCT should be recorded. Moreover, we predicted that the upper motor neurons of SCA2 and SCA3 patients might also be damaged; therefore, we hope that CMCT studies of SCA2 and SCA3 with large samples will be carried out in the future. Finally, CMCT combined with clinical follow-up could be carried out in patients with SCA, especially those with SCA1. The sensitivity and stability of CMCT as an indicator of disease progression could be observed. Our research also had some limitations. For example, some studies were quite old. Additionally, there was heterogeneity among different studies , and the number of studies in subgroup analyses was low after the sources of heterogeneity were excluded. Nevertheless, we did include a larger sample than any previous study, and we were able to derive a meaningful conclusion. We hope that subsequent studies will further expand the data.CMCT prolongation has been observed in different subtypes of SCAs. The difference in CMCT values among different subtypes suggests diverse pathological mechanisms. Our study opens the possibility of performing quantitative analysis and dynamic observation of the extent of damage to the pyramidal tract in SCA patients. As an objective index with which to determine the severity of the disease, CMCT has broad prospects for application, although they need to be confirmed in the future.We conducted this meta-analysis based on the Meta-analysis Of Observational Studies in Epidemiology (MOOSE) guidelines  and the To comprehensively examine the relationship between SCA and CMCT, we conducted a comprehensive search of PubMed and Web of Science using the following keywords: [Mesh] “Spinocerebellar Ataxia” or “spinocerebellar ataxia” or “spinocerebellar ataxias” or “spinocerebellar degeneration” or “spinocerebellar atrophy” or “spinocerebellar degenerations” or “spinocerebellar atrophy” or “dominant ataxia” or “hereditary ataxia” or “SCA” or “olivopontocerebellar atrophy” or “Wadia-Swami Syndrome” or “Machado-Joseph disease” in combination with [Mesh] “Transcranial Magnetic Stimulation” or “transcranial magnetic stimulation” or “TMS” or “central motor conduction” or “central motor conduction time”. For Embase, the retrieval strategy used was as follows: (‘spinocerebellar’/exp OR spinocerebellar) AND (‘ataxia’/exp OR ataxia) AND  AND (‘magnetic’/exp OR magnetic) AND (‘stimulation’/exp OR stimulation). We limited the search to human studies published in English before April 2020. The references of the articles were also comprehensively searched to identify articles that were missed in the database search, and these articles were judged according to the same selection criteria as the search results.We applied several inclusion criteria. First, the study needed to involve SCA patients receiving TMS. The parameters of this group were noted. Second, patient information needed to be complete or nearly complete. Third, an age-matched control group was required, and the parameters of the control group needed to be specified.Studies were excluded if any of the following criteria were met: the sample size was too small (n<5), or the patient information was inadequate; the patients were not diagnosed with SCA by molecular techniques ; TMS wasData were extracted by two independent reviewers , and if there was any disagreement, the data were sent to a third reviewer (L.L.W.) for further exploration. The extracted information was as follows: names of authors, publication year, country, ataxia genotype, sample size, population features , measurement location (upper limbs/lower limbs) and CMCT.To evaluate the risk of bias, an independent analysis of all eligible studies was conducted using the modified Newcastle-Ottawa Scale assessment checklist , which wThe characteristics of each study are summarized in Forest plots were used for the statistical analysis of the data. Differences between two groups are reported as the mean difference . For all tests, P < 0.05 was deemed to be significant. The I² statistic measures the percentage of total variation introduced by heterogeneity rather than by chance. This statistic was used to quantify heterogeneity, with 25%, 50% and 75% indicating low, medium and high degrees of heterogeneity , respectSupplementary Figure 1Supplementary Table 1