Clinical trials, systematic reviews and guidelines compare beneficial and non-beneficial outcomes following interventions. Often, however, various studies on a particular topic do not address the same outcomes, making it difficult to draw clinically useful conclusions when a group of studies is looked at as a whole . This prDevelopment of consensus is required around a set of well-defined, relevant and feasible outcomes for all trials concerning particular obstetric and gynaecologic health conditions, such as preterm birth, incontinence, infertility and menstrual problems. With so many subspecialties involved, this is no easy task. Duplication of effort can be avoided by working with the Core Outcome Measures in Effectiveness Trials (COMET) Initiative, which is working towards core data sets for all medical specialties . ProductJournal editors now invite researchers to take the lead in beginning this work. What will we do as editors to support them and their colleagues? First, we are drawing wide attention to The CROWN Initiative by publishing this editorial in the journals listed below. We shall ensure that the global research community, which includes our many reviewers, is aware of the need for core outcome sets. Submissions which describe development of core outcome sets, if deemed acceptable after peer review, will be effectively disseminated.http://www.comet-initiative.org): “The existence or use of a core outcome set does not imply that outcomes in a particular trial should be restricted to those in the relevant core outcome set. Rather, there is an expectation that the core outcomes will be collected and reported, making it easier for the results of trials to be compared, contrasted and combined as appropriate; while researchers continue to explore other outcomes as well.” We also expect that as new or superior ways of capturing outcomes emerge, core outcome sets will themselves need updating.Our collaboration is not for enforcing harmony at the expense of innovation. To quote from the COMET home page  Initiative by Professor Khalid Khan with permission from the Royal College of Obstetricians and Gynaecologists and John Wiley & Sons Ltd.Reproduced from th May 2014, up to date list available at http://www.crown-initiative.org):Acta Obstetricia et Gynecologica ScandinavicaAmerican Journal of Obstetrics & GynecologyAmerican Journal of PerinatologyArchives of Gynecology and ObstetricsAustralian and New Zealand Journal of Obstetrics and GynaecologyBest Practice & Research: Clinical Obstetrics & GynaecologyBirth: Issues in Perinatal CareBJOG: An International Journal of Obstetrics and GynaecologyBMC Pregnancy and ChildbirthBMC Women's HealthClimactericClinical Obstetrics and GynecologyClinics in PerinatologyCochrane Menstrual Disorders and Subfertility GroupCochrane Pregnancy and Childbirth GroupContraceptionContraception and Reproductive MedicineCurrent Opinion in Obstetrics and GynecologyEuropean Journal of Obstetrics & Gynecology and Reproductive BiologyFertility and SterilityFertility Research and PracticeFetal Diagnosis and TherapyGinekologia PolskaGynecological SurgeryGynecologic OncologyGynecologic Oncology ReportsHuman FertilityHuman ReproductionHuman Reproduction UpdateHypertension in PregnancyInternational Journal of Fertility and SterilityInternational Breastfeeding JournalInternational Journal of Gynecology & ObstetricsInternational Urogynecology JournalJournal of Family Planning and Reproductive Health CareJournal of Gynecologic OncologyJournal of Lower Genital Tract DiseaseJournal of Midwifery & Women's HealthJournal of Obstetrics & GynaecologyJournal of Obstetrics and Gynaecology CanadaJournal of Obstetric, Gynecologic & Neonatal NursingJournal of Ovarian ResearchJournal of Perinatal & Neonatal NursingJournal of Perinatal MedicineMaternal Health, Neonatology, and PerinatologyMaturitasMCN The American Journal of Maternal Child NursingMenopause Review Menopause: The Journal of The North American Menopause SocietyNeurourology and UrodynamicsObstetrics & GynecologyPaediatric and Perinatal EpidemiologyPlacentaPrenatal DiagnosisReproductive HealthThe Breast JournalThe European Journal of Contraception and Reproductive Health CareThe Obstetrician & Gynaecologist (TOG)Twin Research and Human GeneticsUltrasound in Obstetrics & GynecologyWomen’s Midlife HealthThe CROWN Initiative includes the following journals, in alphabetical order (correct on 13


AXAH activity in the endosperm appears soon after the onset of germination and resides in dimers putatively containing two isoforms, AXAH1 and AXAH2. Upon grain imbibition, mobilisation of arabinoxylan and starch spreads across the endosperm from the aleurone towards the crease. The front of arabinoxylan degradation precedes that of starch degradation. Incubation of grains with LAB decreases the rate of loss of both arabinoxylan and starch, and retards the spread of both degradation processes across the endosperm. We propose that starch degradation in the endosperm is dependent on cell wall degradation, which permeabilises the walls and thus permits rapid diffusion of amylolytic enzymes. AXAH may be of particular importance in this respect. These results provide new insights into the mobilization of endosperm reserves to support early seedling growth.Starch degradation in barley endosperm provides carbon for early seedling growth, but the control of this process is poorly understood. We investigated whether endosperm cell wall degradation is an important determinant of the rate of starch degradation. We identified iminosugar inhibitors of enzymes that degrade the cell wall component arabinoxylan. The iminosugar 1,4-dideoxy-1, 4-imino- Starch mobilisation in the endosperm of barley seedlings provides the substrate for early growth following grain germination. The process is understood in detail in terms of control of expression of genes encoding key enzymes, and the structure-function relationships of these enzymes12345It has long been speculated that the rate of starch mobilisation depends not on the rate of synthesis of α-amylase, but on the accessibility of endosperm starch granules to this enzyme following its release into the endosperm89101191516d-glucans (referred to as β-glucan), and 20% of the mass is arabinoxylan (referred to as AX) consisting of β-1, 4-xylan chains with α-l-arabinofuranose substitutions at O-3 and/or O-2 positions of the xylanopyranoside (d-Xylp). Lesser amounts of cellulose, glucomannan, xyloglucan, pectic polysaccharides, callose and arabinogalactan proteins are also present1011In barley grains, about 70% of the mass of endosperm cell walls is -β-d-xylan xylanohydrolase XYN-1  and β-d-xylosidase  increased sharply from about three days post-imbibition (dpi) but endo-xylanase activity  did not start to increase until five dpi  structurally resembles l-Araf substituents of the xylan backbone of AX  is sufficient to inhibit starch degradation when applied to intact, germinating barley grains20d-xylosidase activity in endosperm extracts did not bind to the MonoQ column, but bound to a cation exchange column (MonoS) and eluted as a single peak  . GA alsoWe tested whether LAB affects starch mobilisation through an unanticipated direct effect on starch degrading enzymes. Activity of α-amylase in extracts from barley seedlings was unaffected by LAB  whereas LAB inhibited the activity of purified recombinant maltase (Agl97), the enzyme responsible for the conversion of starch-derived maltose to glucose in the endosperm . HoweverWe used two approaches to investigate whether AX degradation in the endosperm was reduced when grains germinated in the presence of LAB. First, endosperm cell wall preparations from dry grains were treated with bacterial endo-xylanase then fractionated by high-performance anion-exchange chromatography (HPAEC). To identify which peaks corresponded to AX oligosaccharides, some samples were treated with bacterial AXAH prior to fractionation. This treatment resulted in major alterations in the oligosaccharide profile detected by HPAEC, and in the accumulation of free arabinose and xylose . Four peSecond, we used immunofluorescence microscopy to determine the distribution of AX in endosperm cell walls during seedling growth up to six dpi. The monoclonal antibody LM11 recognises unsubstituted and relatively low-substituted xylans and binds strongly to wheat endosperm AXIncubation of grains with LAB substantially retarded the loss of fluorescence from endosperm cell walls. The presence of LAB had little effect on the distribution of fluorescence up to two dpi , but at To visualise the pattern of starch mobilisation in the endosperm, sections from grains at six dpi were stained with iodine solution. At this time point no cellular structures were visible in the endosperm adjacent to the aleurone in grains incubated in water only, indicating extensive mobilisation of all cellular material . Starch Given the apparent importance of AX degradation for starch mobilisation in the endosperm, we investigated further the nature of the enzymes responsible for this process. We used size exclusion chromatography to fractionate extracts of endosperm during seedling establishment (4–10 dpi). A single AXAH activity peak was detected in all samples . Its estAXAH1 and AXAH2 are much more highly expressed in the embryos of germinating seeds than AXAH3 or AXAH4, and AXAH5 transcripts are almost undetectable in this organ21XYN-1 is strongly expressed in the aleurone, whereas XYN-2  or c-Myc (3xcMyc) epitope tags at the C-terminus. Chimeric proteins accumulated only in infiltrated areas of the leaf. They were active enzymes, and they could be specifically and quantitatively precipitated from leaf extracts with affinity matrices for the corresponding epitope tag  affinity matrix. Immunoblot analysis of immunoprecipitated proteins with α-FLAG and anti-HA (α-HA) antibodies showed that both AXAH1-FLAG and AXAH1-3xHA were present  also resulted in a substantial reduction in the ratio of glucose to maltose but had little or no effect on starch degradationIn order to examine the importance of cell wall degradation for the mobilisation of starch in the endosperm of barley grains, we looked for small-molecule inhibitors active against important enzymes of cell wall degradation but not against enzymes directly involved in starch degradation. The iminosugar LAB met these criteria. It inhibited the AX-degrading enzyme AXAH but not the endo-xylanase and xylosidase also implicated in AX degradation. Importantly it did not inhibit key enzymes of starch degradation including α-amylase, the enzyme responsible for the initial attack on starch granules, and β-amylase and limit dextrinase, enzymes which further process the products of α-amylolysis. LAB inhibited the downstream enzyme maltase, but this inhibition is highly unlikely to be responsible for its effects on starch degradation. First, transgenic lines of barley carrying an RNA interference silencing cassette for Despite its lack of direct effect on enzymes important for starch degradation, the application of LAB to germinating barley grains markedly reduced the extent of degradation of both AX and starch over the first ten dpi. Two independent lines of evidence support this conclusion. First, direct measurements of endosperm starch content and the amounts of some specific products of AXAH degradation of endosperm cell walls show that incubation of grains with LAB retarded the rate of loss of both starch and AX during the first six to ten dpi. Second, visualisation of the distributions of starch and AX in endosperm sections revealed that incubation of grains with LAB retarded the spread of a wave of degradation of both polymers that proceeded from the aleurone towards the crease in the first six dpi. These spatial analyses also revealed that major loss of starch from regions of the endosperm was preceded by substantial or complete loss of AX. Taken together, these results are consistent with the view that LAB retards loss of AX from the endosperm, and does so specifically via its inhibitory effect on AXAH, and that the progression of starch degradation across the endosperm follows the loss of AX.The fact that LAB retards the loss of both AX and starch from the endosperm suggests strongly that permeabilisation of cell walls through degradation of cell wall polymers is a prerequisite for starch degradation. It seems highly likely that the diffusion of α-amylase from the aleurone into the endosperm is prevented or strongly retarded by intact cell walls. Previous authors have shown that the temporal and spatial patterns of cell wall and starch degradation are similar and proposed that cell wall degradation may be important for starch degradationd-xylosidase, β-d-glucosidase, α-d-mannosidase and β-d-galactosidase activities, but we have not examined its effects on enzymes that degrade β-glucans. However the clear and strong effects of LAB on AXAH activity in vitro and on AX and starch loss in vivo are consistent with the idea that AX degradation per se has a major impact on cell wall permeability and integrity, and that AXAH is a key enzyme for endosperm mobilisation as a whole. It is widely acknowledged that the arabinose substituents hinder the hydrolysis of the xylan backbone of arabinoxylan during its degradation. Enzymes that remove these substituents act synergistically with endo-xylanase in the complete degradation of the polymer32Our results do not rule out the possibility that LAB retards loss of cell wall integrity by inhibiting more than one key enzyme of cell wall degradation. We showed that it does not inhibit endo-xylanase, β-The known synergistic interactions during arabinoxylan degradation between endo-xylanases and enzymes that remove arabinose substituents of AX prompted us to investigate whether actions of AXAH and endo-xylanase may be coordinated through protein-protein interactions.The two gene products likely to account for AXAH activity in the endosperm, AXAH1 and AXAH2, are 83% identical, but nonetheless have different affinities for AX and arabinan substratesIn line with previous reports1924The consequences of AXAH action for the integrity, permeability and susceptibility to degradation of the endosperm cell wall as a whole are difficult to predict because of the paucity of information about wall structure. Based on the ability of xylanases and arabinofuranosidase to solubilise β-glucans from endosperm cell walls, Bamforth and colleagues proposed that the endosperm cell wall consists of β-glucan sandwiched between outer layers of arabinoxylan424347The extent of degradation of cell walls during the malting of barley (the level of modification) is very important for malt quality. High levels of degradation are generally desirable because degradation products are available for yeast fermentation, and because partially degraded cell walls lead to high viscosity and impeded filtration at later stages of beer productionwww.glycosyn.com) and acarbose from Toronto Research Chemicals Inc. (http://www.trc-canada.com/). DNJ was a kind gift from Dextra Laboratories Ltd or purchased from Callaghan Innovation Research Ltd (http://www.irl.cri.nz/), LAB  was chemically synthesised from d-xylosehttp://www3.gehealthcare.co.uk/). Grains were surface-sterilised2 wells of 25-well sterile plastic plates at 17 °C in the dark.Barley was the UK elite variety NFC Tipple. XyloDNJ was from GlycoSyn (HvAXAH1 and HvAXAH2 (NCBI accessions JQ303075.1 and JQ303076.1 respectivelyHvXYN-1 coding sequencewww.lifetechnologies.com/uk/) and amplified with primers VA36 and VA37. Constructs were cloned into pCR8/GW/TOPO vector, sequenced with primers VA48-VA51 (HvAXAH1), VA55 and VA56 (HvAXAH2), VA38 and VA39 (HvXYN1), and introduced by Gateway LR clonase II-mediated recombination (Life Technologies) in the following vectors: pGWB11 or pGWB14 or pGWB17 for C-terminal FLAG or 3xHA or 3x c-Myc epitope tags respectivelyAgrobacterium tumefaciens strain GV3101. Bacteria were infiltrated into leaves of three-week-old N. benthamiana plants and extracts were prepared two days after infiltration® M2, EZview™ Red anti-HA or EZview™ Red anti-c-Myc affinity matrices (Sigma-Aldrich). Bound proteins were collected and washed as describedPrimers are described in g for 30 min), ammonium sulfate was added to the filtrate and protein precipitating between 20 and 80% saturation was collected by centrifugation as above. The pellet was resuspended in medium A , desalted with Sephadex G25  equilibrated with medium A, then applied to a MonoQ 5/50 GL anion exchange column  equilibrated with medium A, on an AKTA FPLC system . The column was washed with medium A at a flow rate of 1 ml min−1, then eluted with a 40-ml linear gradient of 0–1 M NaCl in medium A. Proteins in the initial wash (unbound MonoQ fraction) were desalted with Sephadex G25 equilibrated with medium B , then applied to a MonoS 5/50 GL cation exchange column equilibrated with medium B. The column was washed with medium B at a flow rate of 1 ml min−1, and eluted with a 40-ml linear gradient of 0–0.5 M NaCl in medium B.For ion exchange chromatography, the roots and coleoptile of 30–40 seedlings at ten dpi were removed and the remaining tissue (referred to as the endosperm), was rapidly frozen and stored at −80 °C until use. Tissue was ground under liquid nitrogen, homogenised in 100 mM MOPS/KOH (pH 7.2), 1 mM EDTA, 1 mM DTT, 10% (v/v) ethanediol, with protease inhibitors  and 2-ml fractions were collected. The column was calibrated with protein standards in the range 29–700 kDa (Sigma-Aldrich) and the void volume determined with Blue Dextran .For size exclusion chromatography, endosperm tissue prepared as above was homogenised in 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, 1 mM DTT, with protease inhibitors . Following centrifugation and filtration through 0.22 μm filters, samples were applied to a Sephacryl S300HR 26/60 size exclusion column  equilibrated with 50 mM Tris/HCl (pH 7.5), 150 mM NaCl, at 1 ml minl-arabinofuranoside (pNPA); endo-xylanase assay, 4-nitrophenyl xylotrioside (OpNPX3); β-d-xylosidase assay, 4-nitrophenyl β–d-xylanopyranoside (pNPX); β-glucosidase assay, 4-nitrophenyl β-d-glucopyranoside (β-pNPG); α-mannosidase assay, 4-nitrophenyl α-d-mannopyranoside (pNPman); β-galactosidase assay, 4-nitrophenyl β-d-galactopyranoside . Agl97 activity was assayed in assays containing 50 mM sodium acetate (pH 4.5), 5 mM CaCl2, 0.6 mM 4-nitrophenyl α-d-glucopyranoside (pNPG)2, 0.005% Triton X-100 (v/v) and 0.1 mg mL−1 pullulanSamples were desalted on Sephadex G25 (coarse) columns equilibrated with 50 mM sodium acetate (pH 5). Activity was measured in assays containing 50 mM sodium acetate (pH 4.5), as described for α-glucosidase® 633 conjugated secondary antibody (Life Technologies), both in 5% (w/v) milk powder in PBS for 1 h. Sections were extensively washed with fresh PBS and overlaid with AF1 glycerol-PBS antifadent solution . For control sections the primary antibody was omitted. Detection of labelled epitopes by confocal microscopy was with a Leica SP5 scan head mounted on a Leica DM6000 upright microscope .The roots and coleoptile of barley seedlings were removed and the remaining tissue immediately immersed in 2.5% (v/v) glutaraldehyde in 50 mM sodium cacodylate (pH 7.2), fixed, treated with osmium tetroxide and dehydratedTransverse sections of barley grains (1 μm) were stained with toluidine blue, counter-stained with Lugol’s iodine solutionAX oligosaccharide generation and fractionation: The husk and embryo were removed from 30–40 dry grains or seedlings and the remaining tissue was pooled, ground under liquid nitrogen and freeze-dried. Samples (100 mg) were prepared as describedTrichoderma viride (Megazyme)57−1. Detection was with an ED40 electrochemical detector (Dionex) with an Au working electrode, and an Ag/AgCl reference electrode in pulsed amperometry mode. Xylose and xylan oligosaccharides (xylobiose to xylohexaose) were identified from the elution profile of standards . Peaks corresponding to AX oligosaccharides were identified through comparison with samples that had been digested with 2 units Aspergillus niger α-d-arabinofuranosidase (Megazyme) at 37 °C for 1 h.Extraction and measurement of starch was as previously describedHow to cite this article: Andriotis, V. M. E. et al. Cell wall degradation is required for normal starch mobilisation in barley endosperm. Sci. Rep.6, 33215; doi: 10.1038/srep33215 (2016).


Marine biofouling is a paramount phenomenon in the marine environment and causes serious problems to maritime industries worldwide. Marine algae are known to produce a wide variety of chemical compounds with antibacterial, antifungal, antialgal, and anti-macrofouling properties, inhibiting the settlement and growth of other marine fouling organisms. Significant investigations and progress have been made in this field in the last two decades and several antifouling extracts and compounds have been isolated from micro- and macroalgae. In this minireview, we have summarized and evaluated antifouling compounds isolated and identified from macroalgae and microalgae between January 2010 and June 2016. Future directions for their commercialization through metabolic engineering and industrial scale up have been discussed. Upon comparing biogeographical regions, investigations from Southeast Asian waters were found to be rather scarce. Thus, we have also discussed the need to conduct more chemical ecology based research in relatively less explored areas with high algal biodiversity like Southeast Asia. An avere Harder  making ae Harder . Biofoule Harder  and is ae Harder . Remediae Harder . To coune Harder ; has lede Harder . The bane Harder , and ince Harder . Howevere Harder . Effortse Harder ; making Benthic marine environments are quite diverse and characterized by extreme competition for light, space, nutrient, and other resources Wahl . Space bNitzschia cf. pellucida  and few microalgal species  have been tested for antifouling activity and several metabolites have been isolated with related bioactivity. The scope of the current review is to cover their activity against maritime fouling and not medical and industrial fouling. In this review, we have summarized the current status of marine algal antifouling compounds and extracts isolated and identified between January 2010 and June 2016. We also present current challenges and future perspectives on development of antifouling strategies from marine algae. Currently, there is an important amount of information on metabolic routes, physiological responses, and algal cultivation, and there are an increasing number of studies on algal genomics  are also known to be a rich source of halogenated secondary metabolites  are known to be good source of bioactive terpenoid derivates, specially meroterpenoids and diterpenoids , Azadinium sp. (Dinophyceae incertae sedis), Dinophysis spp. , Gymnodinium spp. and Karenia spp. , and Prorocentrum spp. , and diatoms as well as Pseudonitzschia spp. . Such diversity is magnified at a rich subspecies level, and in consequence, the heterogeneity of associated phycotoxins and its derivatives are quite variable and big  for feed, and as active ingredients for cosmetics, among others  were isolated from diatoms and dinoflagellates which have demonstrated antimicrobial or antifouling activities  with many islands and rivers flowing into the extended and large continental shelf  are limited in S.E.A. compared to European and North American waters Fig. , neverthAmong the early colonizers, diatoms play a significant role in biofilm development and are able to colonize on even most fouling-resistant surfaces   or cell adhesion complexes    easy, a large production of metabolites with antifouling applications possibly can be achieved. This is supported by the increasing availability of genome level data generated from “omics”-based approaches which can be used as part of RNAi and riboswitch engineering approaches and may lead to enhanced production of antifouling metabolites. At the same time, it can be synchronized with rational biochemical engineering design for large-scale production of marine algal antifouling metabolites using photobioreactor technology. Given that there has been significant technological development in the field of metabolic engineering, one could expect that in coming years, this approach could prove to be useful for development of antifouling paints. However, the genetic improvement of algal strains is a current  challenge till now. Modified strains could overproduce traditional or newly discovered algal compounds and also serve to express specific genes that cannot be expressed into other organisms  fouling control in an environmentally friendly way and (b) successful exploitation of the reported active compounds is yet to be achieved and there is possibly still some way to go for full commercial use of such compounds. While we should still continue looking for such compounds in general and also exploit the untapped potential of S.E.A out of academic interests if not bioprospecting, we highlight the importance of (a) linking up biofilm formation with the diatom genes involved as identification of such genes could form the basis of future biofilm knockout mechanisms and (b) the need to put together screening, genomic, and metabolic engineering studies under one roof for successful interdisciplinary exploitation of such rich biodiversity of marine algal compounds.


Journal of Fungi would like to express their sincere gratitude to the following reviewers for assessing manuscripts in 2015. The editors of the We greatly appreciate the contribution of expert reviewers, which is crucial to the journal’s editorial decision-making process. Several steps have been taken in 2015 to thank and acknowledge reviewers. Good, timely reviews are rewarded with a discount off their next MDPI publication. By creating an account on the submission system, reviewers can access details of their past reviews, see the comments of other reviewers, and download a letter of acknowledgement for their records. In addition, MDPI has launched a collaboration with Publons, a website that seeks to publicly acknowledge reviewers on a per journal basis. This is all done, of course, within the constraints of reviewer confidentiality. Feedback from reviewers shows that most see their task as a voluntary and mostly unseen work in service to the scientific community. We are grateful to our reviewers for the contribution they make.Akins, RobertHay, Roderick J.Nevez, GillesAverill, ColinHaydoura, SouhaNouri, KeyvanBarnhart, KirstinHeinz, Werner J.Orlovich, DavidBhatia, NealHoenigl, MartinPapini, ManuelaBubici, GiovanniIdelevich, Evgeny A.Perlin, DavidChao, Sheau-chiouIorizzo, MatildePfaller, Michael A.Chaturvedi, AshokIson, Michael G.Phillips, Lori A.Choi, Jong SooJaijakul, SirayaPullarkat, VinodCogliati, MassimoKhachemoune, AmorRanque, StéphaneColom, M. FranciscaKouvelis, Vassili N.Reales-Calderón, Jose AntonioDaeschlein, GeorgKranabetter, J. MartyRich, PhoebeDaniel, Carlton Ralph C.Lanternier, FannyRoilides, EmmanuelDemirev, PlamenLauto, AntonioSaville, Stephen P.Enk, Claes D.Lavergne, Rose-AnneStarace, MichelaEsperón, FernandoLetang, EmilioSturtevant, JoyEspinel-Ingroff, AnaLipner, Shari R.Sukdeo, NicoleFranco-Paredes, CarlosLopez-Llorca, LuisSuz, Laura M.Garcia-Diaz, JuliaMack, MeganTeichtahl, AndrewGhannoum, Mahmoud A.Markinson, BryanTorbett, BruceGiashuddin, ShahMartin, RonnyWainwright, MarkGrim, Shellee A.McKenney, JennieWalsh, Thomas J.Gupta, Aditya K.Morgado, Luis N.Weese, ScottHagen, FerryMorton, Charles OliverZaias, Nardo


Journal of Pregnancy has retracted the article titled “Antenatal Steroid Therapy for Fetal Lung Maturation and the Subsequent Risk of Childhood Asthma: A Longitudinal Analysis” . The art


The prosthetic stiffness values of manufacturer recommended stiffness categories varied between prosthetic models (p<0.001). Also, prosthetic stiffness was 10% to 39% less at angles typical of running 3 m/s and 6 m/s (10°-25°) compared to neutral (0°) (p<0.001). Furthermore, prosthetic stiffness was inversely related to height in J-shaped (p<0.001), but not C-shaped, prostheses. Running-specific prostheses should be tested under the demands of the respective activity in order to derive relevant characterizations of stiffness and function. In all, our results indicate that when athletes with leg amputations alter prosthetic model, height, and/or sagittal plane alignment, their prosthetic stiffness profiles also change; therefore variations in comfort, performance, etc. may be indirectly due to altered stiffness.The mechanical stiffness of running-specific prostheses likely affects the functional abilities of athletes with leg amputations. However, each prosthetic manufacturer recommends prostheses based on subjective stiffness categories rather than performance based metrics. The actual mechanical stiffness values of running-specific prostheses (i.e. kN/m) are unknown. Consequently, we sought to characterize and disseminate the stiffness values of running-specific prostheses so that researchers, clinicians, and athletes can objectively evaluate prosthetic function. We characterized the stiffness values of 55 running-specific prostheses across various models, stiffness categories, and heights using forces and angles representative of those measured from athletes with transtibial amputations during running. Characterizing prosthetic force-displacement profiles with a 2 The sd-stance :kleg=FΔInspired by the spring-like nature of running, passive-elastic running-specific prostheses (RSPs) were developed to enable athletes with lower-limb amputations to run. These carbon-fiber devices are attached to the sockets that encompass the residual limbs, are in-series with the residual limbs, and mimic the mechanical energy storage and return of tendons during ground contact. Unlike biological ankles, RSPs cannot generate mechanical power anew and only return 63% to 95% of the stored elastic energy during running . For conAthletes with leg amputations may adapt similar leg spring mechanics as non-amputees by using RSPs that emulate biological lower leg stiffness. Individually, non-amputees adopt a constant ,9,10, meProsthetic manufacturers do not report the stiffness values of RSPs (e.g. in kN/m). Instead, they classify RSPs into predetermined stiffness categories (e.g. categories 1 to 7), which are recommended to users based on body mass and intended activity (slow or fast running) –22. LargTo accurately quantify prosthetic stiffness, it seems obvious to evaluate RSPs using forces and angles indicative of those produced during the respective activity. When athletes with transtibial amputations run, they generate peak vertical ground reaction forces (GRFs) with their affected legs that are 2.1 to 3.3 times body weight at speeds of 2.5 m/s to 10.8 m/s ,25,26. DSince prosthetic stiffness and hysteresis likely affect running performance, we aimed to 1) characterize the force-displacement profiles of RSPs, 2) quantify and compare prosthetic stiffness and 3) hysteresis values across prosthetic models, stiffness categories, and heights using angles and forces that replicate those exhibited during running, and 4) determine whether prosthetic height affects stiffness. Such information will enable accurate and objective comparisons between RSPs, subsequently allowing for potential improvements in prosthetic design, prescription, and athletic performance. Based on the predominant assumption that prosthetic stiffness is constant during running ,16–18; wWe measured GRFs and sagittal plane angles of RSPs relative to the peak resultant GRFs from 11 athletes  with unilateral transtibial amputations while they ran at 3 m/s and 6 m/s on a force-measuring treadmill. Each athlete used their own personal RSP. 3 m/s represents a typical distance running speed  and 6 m/th order low-pass Butterworth filter with a cutoff frequency of 6 Hz  . Our data were comparable because each participant ran both speeds at one lab, and due to the implementation of the same filtering process.We placed reflective markers on the lateral proximal and distal ends of each RSP’s longitudinal axis and measured segment motion during each trial using a motion capture system  at 240 Hz (lab 1) or 200 Hz (lab 2) and implemented a 4MD, USA) . The lonMD, USA) . Four at3) and 6 m/s (α6), and with J-shaped RSPs at 3 m/s (β3) and 6 m/s (β6). When the RSP’s longitudinal axis is parallel to the peak resultant GRF vector, the RSP is at 0°. Positive angles indicate that the proximal longitudinal axis was rotated forward in the sagittal plane relative to the peak resultant GRF vector  and peak resultant GRF magnitudes into our prosthetic testing procedure.We determined the peak GRF magnitude, as well as the average sagittal plane angle of the longitudinal axis for each athlete’s RSP relative to the peak resultant GRF vector from 10 consecutive ground contacts with the affected leg. We assessed the average angles for trials performed with C-shaped RSPs at 3 m/s , Freedom Innovations , and Ottobock  donated a combined total of 55 RSPs for use in our study. We characterized prosthetic stiffness profiles and hysteresis magnitudes from 14 C-shaped Össur Flex-Run prostheses (stiffness categories 3 low– 7 high), 12 C-shaped Freedom Innovations Catapult FX6 prostheses (stiffness categories 2–7), 14 J-shaped Ottobock 1E90 Sprinter prostheses (stiffness categories 1–5), and 15 J-shaped Össur Cheetah Xtend prostheses (stiffness categories 2–7)  Table 1Table 1. 3, and α6, as well as under each J-shaped RSP at 0°, β3, and β6  . We also, and β6 . We applTo determine the peak GRF magnitude applied on each RSP, we considered the heaviest manufacturer recommended body weight for each prosthetic stiffness category, then multiplied it by 3.0 to replicate the upper limit of peak GRFs typically produced by affected legs while running 3 m/s , and by To determine the effect of prosthetic height on the stiffness of C-shaped RSPs, we tested the Catapult and Flex-Run prostheses at 38.2 cm and 69.7 cm by altering the aluminum pylon height. To determine the effect of height on the stiffness of J-shaped RSPs, we tested the 1E90 Sprinter prostheses at 25.0, 31.5, and 38.0 cm, and the Cheetah Xtend prostheses at 31.5, 38.0, and 41.5 cm. Prosthetic height was measured vertically from the ground to the base of our height adjustment attachment in an unloaded state . We chos2) for linear and curvilinear characterizations of the applied force relative to the vertical displacement for each 3-cycle trial. Next, we averaged R2 values within and across trials for a given prosthetic model, stiffness category, height, and testing angle combination. Furthermore, we calculated average prosthetic stiffness for each model across stiffness categories using the force-displacement function during simulated running conditions.To characterize prosthetic stiffness, we calculated the average coefficients of determination (RF is the applied force as a function of the change in prosthetic height (h) and peak change in prosthetic height (H) of the corresponding cycle. Hysteresis was averaged for each 3-cycle trial, and averaged across trials of the same prosthetic model, stiffness category, height, and testing angle. We measured prosthetic stiffness and hysteresis with the respective manufacturers supplied rubber sole. We also measured the stiffness and hysteresis of the highest stiffness category from each model at 0° without the rubber sole.For every cycle, we calculated hysteresis as the ratio of energy lost during recoil relative to the energy stored during compression, then expressed it as a percentage:2 values from linear and curvilinear force-displacement functions across prosthetic models and to compare the manufacturer recommended stiffness across prosthetic models for athletes at body masses of 55 kg to 100 kg in 5 kg increments using the average angles and peak applied force magnitudes produced at 3 m/s (α3 and β3) from the C- and J-shaped RSPs, respectively. We also used paired two-tailed t-tests to compare the prescribed stiffness of different prosthetic models for athletes at body masses of 55 kg to 100 kg in 5 kg increments using the average angles and peak applied force magnitudes produced at 6 m/s (α6 and β6) from the C- and J-shaped RSPs, respectively. The recommended stiffness values for J-shaped RSPs were calculated using the tallest mutual height (38 cm).We used paired two-tailed t-tests to compare average RMoreover, for C-shaped RSPs, we used linear mixed models to compare 1) prosthetic stiffness and 2) hysteresis for each prosthetic model across stiffness categories, testing angles, and interaction effects. For the J-shaped RSPs we included prosthetic height as an independent variable and used two linear mixed models to compare 1) prosthetic stiffness and 2) hysteresis for each prosthetic model across stiffness categories, testing angles, and heights, in addition to their interactions. We performed paired two-tailed t-tests to assess the influence of the prosthetic sole on stiffness and hysteresis. We carried out our statistical analyses using R-studio  software. Significance was set at p<0.05. When applicable, we implemented the Bonferroni correction to account for multiple comparisons.3 = 15.0°, α6 = 10.0°, β3 = 20.0°, and β6 = 25.0°.When participants used C-shaped RSPs to run 3 m/s, the average angle of their RSP’s longitudinal axis relative to the peak resultant GRF was 15.1° ± 4.8° and the mean peak resultant GRF was 2.5 ± 0.3 times body weight. At 6 m/s the average angle was 10.0° ± 4.2° and the peak resultant GRF was 2.7 ± 0.3 times body weight. When participants used a J-shaped RSP to run 3 m/s, the average angle of their RSP’s longitudinal axis relative to the peak resultant GRF was 20.9° ± 8.9° while the average peak resultant GRF was 2.6 ± 0.3 times body weight. At 6 m/s, the average angle was 24.2° ± 9.3° and the average peak resultant GRF magnitude was 2.8 ± 0.3 times body weight. Since our custom base was constructed to rotate in incremental steps, we used the following values for RSP testing: αnd degree polynomial explained 4.4% more of the variance than a linear function using angles indicative of 3 m/s and 6 m/s (p<0.001) . We did not explore functions beyond a 2nd degree polynomial due to its impeccable fit (average R2 = 0.998).Overall, characterizing the slope of the force-displacement curves with a 2p<0.001) . At a teUsing the peak resultant GRFs and angles produced at 3 m/s, the actual stiffness of the manufacturer recommended Cheetah Xtend, which is prescribed based on user body mass, was 4% to 15% stiffer than the Flex-Run (p<0.001), 7% to 19% stiffer than the Catapult (p<0.001), and 20% to 28% stiffer than the 1E90 Sprinter (p<0.001) prostheses across matched user body masses . Using tnd order polynomial equations for each prosthetic model and stiffness category (S1-S4) so that prosthetists can predict an athlete’s prosthetic stiffness from the amount of force they apply on the ground and/or prosthetic compression. For those unable to quantify force magnitudes or compression, and because of the relatively linear force-displacement relationships (average R2 = 0.956), we also report average linear stiffness values . For every 1° increase in testing angle, the hysteresis magnitude decreased 0.04% (p<0.001). The average hysteresis for J-shaped RSPs across conditions was 4.28% (SD: 0.65%), which was lower than that of the C-shaped RSPs (p<0.001). Furthermore, testing angle affected the hysteresis of J-shaped RSPs (p<0.001), while height had no effect (p = 0.215). For every 1° increase in testing angle, the hysteresis of the 1E90 Sprinter and Cheetah Xtend prostheses decreased 0.01% and 0.08%, respectively (p<0.001). Additionally, removing the rubber soles from C- and J-shaped RSPs reduced the hysteresis magnitudes by 42% (p<0.001).While controlling for prosthetic height, every 1° increase in testing angle decreased the stiffness of the Flex-Run and Catapult prostheses by 0.41 kN/m (p<0.001) and 0.79 kN/m (p<0.001), respectively . Every 12 = 0.956), a linear function did not fit quite as well as a 2nd degree polynomial function (p<0.001), leading us to partially reject our initial hypothesis. Contrary to the notion that prosthetic stiffness is invariant during running [Despite well-characterizing the force-displacement relationships of the RSPs  by 10% to 39% compared to using angles utilized by athletes with transtibial amputations while running at 3 m/s and 6 m/s. Previous studies have tested the stiffness of RSPs at 0° , and 30°We reject our second hypothesis; manufacturer recommended prosthetic stiffness varies across models for a given user body mass and activity. Additionally, we compared manufacturer recommended prosthetic stiffness during running at 6 m/s versus at 3 m/s. At a given body mass (prosthetic height of 38 cm), the manufacturer recommended 1E90 Sprinter prostheses were 11% stiffer at 6 m/s compared to 3 m/s across a 45 kg span in user body mass (p = 0.003). Also, the recommended Catapult prosthetic stiffness increased 32% due to a greater recommended prosthetic stiffness category and reduced angle between the RSP and peak resultant GRF . ConversSince testing angle affected hysteresis, we also reject our third hypothesis stating that prosthetic hysteresis would be invariant across testing conditions. Intriguingly, RSPs dissipate less energy when their proximal end is rotated forward with respect to the applied force. Future studies are needed to examine prosthetic designs and decipher why RSPs display less hysteresis when rotated forward. Due to the importance of mechanical energy return on running and sprinting performance ,5, the dMoreover, prosthetic hysteresis was 42% lower when we removed the rubber soles, indicating that the rubber soles were responsible for almost half of the dissipated energy. Athletes with leg amputations should use soles with minimal damping to maximize the mechanical energy return of RSPs. In addition to the sole, energy dissipation probably occurs at the residual limb/socket interface. To our knowledge, no study has quantified the mechanical behavior of the residual limb and socket interface while running. Improving socket design by enhancing the connection between athletes and their RSPs may allow better utilization of the returned mechanical energy and potentially improve running performance.Pylon height does not affect the stiffness of C-shaped RSPs; therefore, we reject our final hypothesis. The aluminum pylon of C-shaped RSPs has an annular section (i.e. an empty cylinder) and appears less prone to bending due to the perpendicular components of the applied compression forces, and due to a higher area moment of inertia  comparedWe assumed that the C-shaped RSPs were perpendicular to the respective pylons. Yet, the sagittal plane RSP-pylon alignment may have been slightly altered due to individual preference, thus our reported angles between the C-shaped RSPs and resultant GRF vectors may have been over/underestimated by a few degrees. We collected prosthetic angles and peak resultant GRFs from a cohort of exceptional athletes with unilateral transtibial amputations at 3 m/s and 6 m/s. Conceivably, less athletic individuals with amputations, or athletes with different amputation levels may not utilize the same prosthetic angles and/or generate the same resultant GRFs compared to those exhibited by our participants, and consequently prosthetic stiffness may differ. For example, athletes with transfemoral amputations with pylons connecting their RSPs to their sockets can use our reported values at 0°, as it is a fair approximation of their RSP-peak GRF angle to determine the prosthetic stiffness and hysteresis.Our methodology does not account for the rotation of the RSP with respect to the resultant GRF throughout ground contact. It may be that RSPs are stiffer at initial and terminal ground contact than at mid-stance due to a smaller angle between the RSP and resultant GRF vector. On the other hand, as applied force accrues RSPs become stiffer, implying that RSPs are stiffest at mid-stance. The influence of angle and force may counteract each other, exhibiting a constant prosthetic stiffness throughout stance; perhaps a deliberate design choice of prosthetic manufacturers. Future studies are warranted to include a rotational component to the mechanical stiffness testing of RSPs. Furthermore, we tested our RSPs with a loading rate (100 N/s) that is much lower than that recorded during running . HowWe assessed prosthetic stiffness and hysteresis across a wide range of models, stiffness categories, and heights, at forces and angles that simulate those exhibited by athletes with transtibial amputations running at 3 m/s and 6 m/s. We found that the force-displacement profiles of RSPs are curvilinear, indicating that prosthetic stiffness varies with the magnitude of applied force. Yet, a linear force-displacement characterization is strongly predictive. We also found that manufacturer recommended prosthetic stiffness varies between models, and that the height of J-shaped RSPs is inversely related to stiffness. Moreover, we provide evidence that prosthetic stiffness is much greater at 0° than at angles representative of those that occur during running.When athletes with leg amputations change prosthetic models, height, and/or sagittal plane alignment, prosthetic stiffness also changes; therefore variations in comfort, performance, etc. may be indirectly due to altered stiffness. We propose that prosthetic stiffness should be assessed under conditions that simulate the demands of the respective activity, and that manufacturers should provide the stiffness values of each RSP at specific heights. Until then, our study provides reference for the stiffness values of various prosthetic models across multiple stiffness categories and heights, and provides a foundation for future research to understand the potential effects of prosthetic stiffness on performance during distance running and sprinting.S1 TableThe equations indicate prosthetic displacement in meters (h) used to calculate the applied force in kN. Stiffness equals applied force divided by displacement. a and b are constants. All prostheses were tested with the manufacturer supplied sole, with the exception of stiffness category 7 High No Sole.(DOCX)Click here for additional data file.S2 TableThe equations indicate prosthetic displacement in meters (h) used to calculate the applied force in kN. Stiffness equals applied force divided by displacement. a and b are constants. All prostheses were tested with the manufacturer supplied sole, with the exception of stiffness category 7 No Sole.(DOCX)Click here for additional data file.S3 TableThe equations indicate prosthetic displacement in meters (h) used to calculate the applied force in kN. Stiffness equals applied force divided by displacement. a and b are constants. All prostheses were tested with the manufacturer supplied sole, with the exception of stiffness category 5 No Sole.(DOCX)Click here for additional data file.S4 TableThe equations indicate prosthetic displacement in meters (h) used to calculate the applied force in kN. Stiffness equals applied force divided by displacement. a and b are constants. All RSPs were tested with the supplied sole from the Össur Flex-Run prostheses, with the exception of stiffness category 7 No Sole.(DOCX)Click here for additional data file.


The following scientists assisted the journal by reviewing manuscripts during the period October 1, 2015 to October 31, 2016. We gratefully acknowledge their critical evaluation and assistance in selecting articles for publication.


Chu, who upheld great standards of science during very difficult times in China. His contributions to the genetic characterisation of influenza viruses were extraordinary.”—Professor Peter Palese, a member of the National Academy of Sciences and expert in the field of RNA viruses, wrote this when he received the obituary notice of Dr. Chu in 1998.“He was not only a great scientist and teacher, but also a remarkable human being who contributed significantly to openness in science and international relations.”—Professors Graeme Laver and Robert Webster, famous virologists at the Department of Virology and Molecular Biology , wrote an obituary for Dr. Chu in 1999  was an internationally renowned virologist, one of the most important founders of molecular virology and a pioneer of biologicals in China Fig. . He was Dr. Chu was born on September 12, 1917, in Yixing County, Jiangsu Province. He graduated from Shanghai Medical College in 1939 and was awarded a Ph.D. by Cambridge University (UK) in 1948. He worked for the World Influenza Centre at the National Institute for Medical Research at Mill Hill (London) from 1948 to 1950. After returning to China in 1950, he worked successively at the National Vaccine and Serum Institute (Beijing), National Institute of Biologicals in Changchun and the Chinese Academy of Preventive Medicine (CAPM)  cells, assuming directorship of the project. After more than 10 years of hard work, he and his colleagues finally succeeded in developing a genetically engineered hepatitis B vaccine expressed in CHO cells, which was the first genetically engineered HBV vaccine produced in CHO cells  at the Institute of Virology (CAPM). In 1981, he published a paper titled Virology (Bing du xue bao), Dr. Chu made specific comments on each article accepted by this journal, such as on academic issues, English usage and illustrations. He made the final decision to accept or reject a manuscript after peer review based on academic quality , Acta Microbiologica Sinica and the Chinese Journal of Epidemiology.Based his distinguished contributions to virology and biologicals, Dr. Chu was honoured with the National Science Conference Award, the First-Class Award for Scientific Advancement by the State, the Third-Class Prize in Natural Science of the State, and the First-Class Award for Scientific Advancement by the Ministry of Health, and the Ho Leung Ho Lee Scientific Advancement in Medical Science Award. he was invited to be an adviser to the international journal Dr. Chu was a truly international Chinese virologist who made significant contributions to solving key problems in viral disease control in China. Dr. Chu has died, but his spirit and style are eternal. We, his former colleagues and students, penned this memorial on the occasion of the centennial of Dr. Chu’s birthday. We try always to emulate Dr. Chu’s spirit, courage, enthusiasm, meticulousness and dedication, and to be worthy recipients of his legacy.


Large radial tears of donor Descemet membrane (DM) during the preparation of Descemet membrane endothelial keratoplasty (DMEK) grafts can make the trephination stage impossible because of small graft diameter. This results in irregular-edged grafts. In this study, we report two pseudophakic bullous keratopathy patients who underwent DMEK surgery with irregular-edged Descemet membrane (DM) grafts. Main outcome measures were preoperative and postoperative 1-, 3-, and 6-month best corrected visual acuity (BCVA), endothelial cell density (ECD) and central corneal thickness (CCT). Intraoperative and early postoperative complications were also evaluated. Both irregular-edged grafts were successfully implanted into the anterior chamber, unfolded, and attached to the posterior corneal stroma. Patients’ BCVA at 6 months was 1.0  and 0.6  respectively. Decrease in ECD at the last visit was 27% and 25%. CCT decreased from 723 μm and 850 μm to 530 μm and 523 μm, respectively. No intraoperative complications occurred except for the large radial Descemet membrane graft tears that developed during donor DM stripping. None of the cases needed a rebubbling procedure postoperatively. We have demonstrated that irregular-edged DM grafts can be successfully implanted for DMEK surgery with good clinical outcomes. Standardized techniques for graft preparation, surgical instruments designed for endothelial keratoplasty, and accumulating experience over time have led to a significant reduction of tissue loss. However, as the donor-related risk factors for failure in donor tissue preparation have not been clearly determined, the potential risk for radial DM tearing still exists. By pulling the torn flaps peripherally and skipping the trephination stage of the graft preparation technique described by Melles et al.,4 these irregular-edged grafts can still be successfully implanted. In this study, we describe the clinical results of two eyes with pseudophakic bullous keratopathy treated with DMEK with irregular-edged DM graft.Descemet membrane endothelial keratoplasty (DMEK) is the latest refinement of endothelial keratoplasty procedures. Providing an exact anatomical replacement of only what is removed, it gives the possibility of excellent visual acuity with shorter healing time as well as minimal risk of immunological rejection.Case 1 was a 72-year-old female patient with a 1-year history of pseudophakic bullous keratopathy in the right eye. Intraocular lens was in the bag and posterior capsule was intact. Best corrected visual acuity (BCVA) was counting fingers from 1 meter (20/2000) with a central corneal thickness (CCT) of 723 µm. Case 2 was a 48-year-old female patient who underwent primary suturation after a penetrating corneal injury to her left eye 2 years earlier. She had scleral fixation of an IOL and trabeculectomy for glaucoma 1.5 years earlier. She had endothelial decompensation in the left eye with a BCVA of hand motions (20/20000). Her CCT was 850 µm with a central corneal scar. Both cases were contact lens-dependent and were using 5% hypertonic ophthalmic solutions.2, respectively) and stored in corneal storage medium  at 4 °C. Graft preparation was done preoperatively in the operating room by applying the technique described in detail by Melles et al.4 Cornea-scleral buttons were mounted endothelial side up on a holder. Descemet membrane–endothelial complex (DEC) was dissected gradually from the periphery to the center starting with a hockey stick knife. In both cases, large radial tears formed during stripping because of focal adhesions between DM and stroma. Radial tears were manipulated by pulling the flap peripherally, forming an irregular edge with no radial tears. This technique allowed preparation of irregular-edged, non-uniform grafts. In our standard technique, stripped DEC surrounded by a 360-degree trabecular meshwork ring is transferred on a soft contact lens and trephined to obtain a 9-9.5-mm regular-edged circular graft. However, since both grafts were smaller than 9 mm in the largest dimension, trephination was not possible. For both cases, the irregular-edged grafts were formed into a DEC roll with the endothelium on the outside and stored in saline until the recipient’s cornea was prepared.Donor cornea-scleral buttons were obtained less than 24 h postmortem  2450 and 2530 cells/mm5 After making three 23-gauge and one 3.2-mm keratotomies, the DM was scored at a radius of 9.5 mm with a reverse Sinskey hook  and removed from the anterior chamber under complete air fill. Then 0.1 mL of acetylcholine chloride  was used to induce myosis in case 2 to reduce the risk of posterior dislocation of DEC roll. The DEC roll was injected into the recipient’s anterior chamber using a glass injector  and oriented endothelial side down by indirect manipulations with air and balanced salt solution. The graft was then uncurled and centered with a combination of corneal dome compression and sweeping of the corneal surface with cannulas. We ensured the graft covered the visual axis. Once the graft was centered, an air bubble was injected underneath the graft and left for 30 minutes under complete air fill. At the end of the procedure, 75% air fill was left , and 0.5% moxifloxacin drops 5 times daily for 2 weeks.The postoperative period was uneventful for both patients, with no DEC detachments requiring rebubbling. Both eyes had improved corneal clarity  and incrDecrease in endothelial cell density at 6 months was 27% and 25% respectively. Central corneal thickness decreased from 723 µm and 850 µm before surgery to 530 µm and 523 µm respectively at postoperative 6 months . PreoperIn the immediate postoperative period, both cases had focal corneal edema limited to the areas of bare corneal stroma. These edematous areas corresponded with the denuded recipient stroma with no DEC because of the irregular graft shape. Edema resolved by the postoperative 6-month visit.7 However it can be complicated with large radial tears, making the trephination and usage of DEC graft impossible. Especially if the graft is prepared by the surgeon prior to surgery in the operating room, radial tear risk increases due to time limitations and surgical stress. To reduce the surgical stress, the surgeon can use eye bank-prepared donor tissue or prepare the tissue days before the surgery. It has been shown that eye bank-prepared grafts and surgeon-prepared grafts do not differ in terms of graft survival outcomes. Both preparation methods have a 5% graft preparation failure rate due to strong adhesions between DEC and stroma.8 Although grafts prepared by the eye bank might have an advantage in decreasing the surgical stress, eye bank-preparation is more expensive compared to preoperative preparation. Even when the surgeon is preparing the graft days before surgery, the use of an extra corneal storage solution increases the total cost of the procedure. Radial tears in DEC grafts that are formed during preparation might increase in size during implantation and unfolding of the graft, resulting in dehiscence of the graft postoperatively. DM graft preparation for DMEK surgery has been standardized previously with very low rates of tissue damage due to preparation.10 Since the risk of losing tissue is still the biggest concern of DMEK graft preparation, the modification we propose can be a salvage method for using grafts with large radial tears. This complication can be managed with a modification of standardized donor tissue preparation technique: rescuing the radial tears by pulling the flap peripherally, skipping the trephination phase, and implanting the irregular-edged graft. Recently, two studies have shown that partial DEC grafts can be implanted and may yield good clinical outcomes.11With irregular-edged grafts, large areas of denuded stroma with edema can be seen in the first postoperative months. Spontaneous resolution of focal edematous areas without DEC may be attributed to the migration of donor and/or recipient endothelial cells onto the denuded stroma.Although two cases may not be enough to evaluate the potential clinical outcomes of irregular-edged grafts, our results seem promising to salvage grafts with large radial tears. These irregular-edged, non-uniform DM grafts might be successfully implanted for DMEK surgery with the potential for favorable clinical outcomes.


Translational Hematology, a new section of the Journal of Translational Medicine. This section aims to support translational research in the discipline of Hematology. It will encompass research in the fields of Red Blood Cells, White Blood Cells, Thrombosis and Hemostasis, Transfusion Medicine, and Hematopoietic Progenitor Cell Transplantation.I am excited to announce the launch of Translational Hematology to not only bring together investigators in translational hematology but also to provide the framework for interaction between investigators from multi-disciplines within one single journal.Hematology is the study of normal hematopoiesis, and the pathology and diseases of the blood, lymphatic system, and bone marrow. It has always been a discipline that has incessantly promoted translational research, especially in the last two decades. Studies in red blood cells have progressed from the simple microscopic examination of morphology and physical evaluation of rheology to the genetic therapy of inherited red blood cell disorders and modern iron chelating agents. Therapy for various hematologic malignancies has also evolved from the use of non-specific chemotherapeutic agents to selective molecular and immune targeting. The availability of new recombinant coagulation factors for congenital bleeding disorders, novel oral anticoagulants for thrombotic disorders, and artificial blood products instead of homologous blood will all provide the opportunity for interesting and clinically relevant translational research that will change clinical practice. Selective T cell depletion to prevent graft-versus-host disease without compromising the graft-versus-tumor effects, ex vivo graft engineering and hematopoietic progenitor cell expansion, and the safer and wider applicability of haplo-identical hematopoietic progenitor cell transplant are just a few of the many active translational research activities ongoing. The recent discovery of the effects of the intestinal microbiota on both sickle cell vaso-occlusive crisis and the development of graft-versus-host disease following allogeneic hematopoietic progenitor cell transplant further supports the interfacing of hematology with other disciplines. The vast range of translational research in hematology, therefore, provides the basis for the establishment of a specially dedicated forum such as Translational Hematology section of the Journal of Translational Medicine aims to provide the vehicle to disseminate research data on benign and malignant hematology, and transfusion medicine. Since hematology is a very diverse discipline that includes cellular biology, molecular biology, and transplantation immunology, we expect that the Translational Hematology section will be a very dynamic section of the Journal of Translational Medicine. The section should, therefore, be of great interest to investigators in both hematology and many other disciplines. We are particularly interested in promoting research results from novel pre-clinical and early clinical translational efforts. New ideas, hypothesis-driven perspectives, and reviews on any particular aspects of hematology are also highly desirable and encouraged because we believe that such contributions will provide the platform for further discussions and hypothesis-testing to hasten our efforts to seamlessly bring successes from bench to bedside.The Journal of Translational Medicine to provide rapid publications, open-access, and high standard peer-review process should provide an ideal platform for a highly interactive and timely discussion among investigators. The Translational Hematology section is proud that its Editorial Board is made up of many high caliber basic and physician scientists of top class scientific and translational reputation in their respective areas of hematology. This will definitely ensure that every published manuscript is of the highest quality and internationally competitive. On behalf of the Editorial Board, I look forward to receiving your contributions to this forum.The mission of the Translation Hematology.Seah H Lim MD PhD FRCP (Lond.) FRCPath. FACP, Section Editor,


Process parameters and post-processing heat treatment techniques have been developed to produce both shape memory and superelastic NiTi using Additive Manufacturing. By introducing engineered porosity, the stiffness of NiTi can be tuned to the level closely matching cortical bone. Using additively manufactured porous superelastic NiTi, we have proposed the use of patient-specific, stiffness-matched fixation hardware, for mandible skeletal reconstructive surgery. Currently, Ti-6Al-4V is the most commonly used material for skeletal fixation devices. Although this material offers more than sufficient strength for immobilization during the bone healing process, the high stiffness of Ti-6Al-4V implants can cause stress shielding. In this paper, we present a study of mandibular reconstruction that uses a dry cadaver mandible to validate our geometric and biomechanical design and fabrication  of NiTi skeletal fixation hardware. Based on the reference-dried mandible, we have developed a Finite Element model to evaluate the performance of the proposed fixation. Our results show a closer-to-normal stress distribution and an enhanced contact pressure at the bone graft interface than would be in the case with Ti-6Al-4V off-the-shelf fixation hardware. The porous fixation plates used in this study were fabricated by selective laser melting. Additive manufacturing (AM), also known as 3D printing, is useful for creating complex parts directly from Computer Aided Design (CAD) software. In general, all AM processes create physical parts directly from CAD data by adding material in successive layers. This layer-by-layer production process makes it possible to fabricate structures with engineered porosity. Current 3D printing devices used for the additive manufacturing of metals create layers by sintering or melting metal powders with a laser or an electron beam. These powder-bed technologies, such as Selective Laser Sintering (SLS), Selective Laser Melting (SLM), and direct metal laser sintering are the most common AM processes for AM fabrication of metal parts ,2.NiTi is the most common shape memory alloy (SMA) showing unique functional properties, i.e., shape memory and superelasticity behavior ,5. PorouHabijan et al.  have shoIt should be pointed out that the as-fabricated Ni-rich NiTi needs post-process heat treatments  to acquire superelasticity behavior. In this study, we have completed a case study by using AM fabricated stiffness-matched porous NiTi skeletal fixation plates that would be useful in mandibular reconstructive surgery.The standard of the care reconstructive surgery to repair a segmental defect of the mandible (lower jaw) involves the use of bone grafts, metallic fixation plates, and metallic screws to restore the mandible’s normal appearance and function  . SurgeonThe high stiffness of Ti-6Al-4V fixation plates and screws can cause abnormal stress distribution throughout the mandible. Over time, this abnormal stress distribution may result in stress concentration in the fixation hardware and screws as well as stress shielding  of the cortical surface of the grafted bone and the host mandible. Stress-shielded bone may resorb, which may then lead to implant failure . LiteratUsing less stiff NiTi fixation plates may reduce stress shielding of grafted bone and elevate the compression force at the graft-host bone interfaces and subsequently enhance revascularization and healing of the grafted bone ,35,36. FThe design of a patient-specific skeletal fixation device starts from Computed Tomography scan data (CT scan data) of the patient. CT scans use X-ray images taken from different angles to produce tomographic images . The varThis section highlights the development, calibration, and validation of a comprehensive model-based NiTi fixation hardware design work path as summarized in A CT scan  of a dry cadaveric human mandible was created to produce a 3D CAD file of all mandible components including cortical bone, cancellous bone, and the teeth. This device utilizes a 64-row Quantum detector, which enables the Aquilion CT scanner to acquire 64 simultaneous slices of 0.5 mm thickness with each 400 ms gantry revolution, resulting in precise isotropic imaging of the object. The gap between the teeth and the cortical bone was graphically filled with periodontal ligament tissue in ABAQUS . For this study, a segment of the left half of the mandible bearing M1-3  was graphically resected with a length of 40 mm. To simulate a high fidelity reconstructive surgery, a double barrel fibular graft, inferior fixation plate, superior distal fixation plate, superior mesial fixation plate and screws were modeled and implanted in the resected region using SOLIDWORKS .As shown in All of the mandible graphical components were meshed in Hypermesh  with 4-node tetrahedral elements (C3D4), which represent 3D, Solid, Tetrahedral, Deformable element type. The optimized number of elements is reported in All meshed parts were assembled, and constraints between screw-fixation plate, screw-host mandible, screw-fibular graft, teeth-ligaments and host mandible-ligaments were defined as tie constraints. It should be noted that the friction factors of 0  and 1  are considered for simulation of surface to surface contact between host mandible and the fibular graft.The bite force created by masticatory muscles is the greatest source of stress that is usually applied to the mandible ,51,52. OCase (A): The simulation of the highest bite force after surgery (without applying any pretension to the fixation hardware). Since the reconstruction surgery causes a reduction in chewing power, all the muscle force values in this study were assumed to be of 60% of Korioth et al.’s  values fCT-scan data were used in the design of the fixation plates . This eliminates the need to bend fixation plates to fit the mandible during the surgery. In this study, the local lingual side curvature of each fixation plate was designed based on the patient’s mandible shape. The overall shape of the fixation plate was also designed to accommodate the insertion of screws at regular intervals and using commonly used thread geometries. The nine-hole inferior fixation plate has a buccolingual thickness of 1.5 mm, a mesiodistal length of 78 mm and a superoinferior width of 4 mm. Each of the two three-hole superior fixation plates has the dimensions of 1 mm × 18 mm × 2.8 mm, in buccolingual, mesiodistal and superoinferior direction, respectively. Six bicortical screws  and four unicortical screws  were designed to fix the inferior fixation plate and both superior plates, respectively , to the In addition to making sure that the fixation plate geometry ensures a good fit, directly on the surface of the host mandible and grafted fibular bones, this project also endeavors to ensure that the stiffness of this plate, and the screws holding it to the mandible and graft segments, have a stiffness that is matched with the stiffness of the cortical bone to which they are attached. The stiffness of cortical bone in the adult mandible varies between 10–31.2 GPa  [1]. TheThe SLM-fabricated, dense NiTi samples still had a higher stiffness  tests to obtain the required material properties ,57,58,59ur group . Those pur group ,63,64,652O), aged , and then water quenched. To perform solution annealing, Lindberg/Blue M BF514541 Box furnace was used. To avoid oxidation of the samples, they were placed in an argon-filled quartz ampules. For determining the transformation temperatures (TTRs), a Perkin-Elmer Pyris 1 DSC with the heating/cooling rate of 10 °C/min in a nitrogen atmosphere was used. The samples were loaded up to 800 MPa  and unloaded using a 100 kN MTS Landmark servo-hydraulic test platform . The strain rate of 10−4·s−1 was employed during loading, whereas unloading was performed under force control at a rate of 100 N·s−1. An MTS high-temperature extensometer was used to obtain strain measurements [After the fabrication process, the cylindrical samples were solution annealed  subroutine based on the microplane theory ,68,69 wa2O) and aging  were performed.FEA was used to determine the stiffness needed to stiffness-match the 3D printed fixation hardware. The final design of the porous NiTi fixation plates consisted of a desired level percentage of porosity, pore shape and size, and general dimension of the plate. The resulting CAD file of the designed NiTi fixation plate was used on the SLM machine for fabrication. To induce superelasticity, the same heat treatment procedures including solution annealing  = 12.6 MPa and r > 0.98, p < 0.005, RMSE = 38.3 MPa for the simulation of loading and unloading responses, respectively).The same UMAT was then used to simulate the behavior of porous superelastic parts. r > 0.99, p < 0.0005, RMSE = 2.8 × 10−6 (%) and r > 0.99, p < 0.0005, RMSE < 6.42 × 10−6 (%) for the Buccal and Lingual sides of mandible cortical bone in the molar region, respectively).An experimental study by Ichim et al. group  is compaThe results of our FEA models are presented at two different time points: (1) immediately following surgery, i.e., when bone fracture healing is occurring and the hardware is bearing most of the strain from chewing and other mandibular activities; and (2) post-healing where stiffness-matched fixation hardware is expected to increase the loading of the grafted bone over traditional, Ti-6Al-4V hardware, and drive the remodeling process ,76. One Graft unvascularization is counted as one of the reasons for the mandibular reconstructive surgery failure. It is often caused by the relative movements, which are decreased as the contact pressure in the surfaces between the graft and the host bone increases . For theIn order to study the stress shielding effect, the von Mises stress distribution in the MRS was investigated using the current standard of care (model 2) and the mandible reconstructed with stiffness-matched NiTi (model 3). Both models were simulated in three different loading cases: case (A), applying highest biting force ; case (B), applying a pure pretension (100 N) on each fixation; case (C), applying a pretension while the mandible is under highest biting force.As As mentioned, a Young’s modulus of 12 GPa for NiTi fixation plates can be achieved by applying a specific percentage and type of porosity . The unit cell of the structure is made out of three perpendicular cylinders with a diameter of 1.00 mm. As a step toward the manufacturing of patient-specific fixation plates, we used a previously healthy, dried cadaver mandible. To simulate the post healing period following reconstructive surgery. Two vertical fractures were created on the left M1-3 section of the mandible. Then a set of three porous NiTi fixation plates similar to the finite element model were designed based on the geometry derived from the CT-scan image of the dried mandible. Subsequently, they were fabricated using a Phenix PXM SLM machine.Additive manufacturing methods have become popular for producing medical devices with high quality. The commonly used material for metallic implants, i.e., Ti-6Al-4V, presents high modulus of elasticity of 112 GPa, while the range of Young’s modulus of the bone varies in the range of 10–31.2 GPa. This high stiffness may result in implant failure due to the stress shielding effect, which results in un-vascularized bone. NiTi is a good candidate for biomedical devices because of lower stiffness (37 GPa), and the superelasticity behavior similar to that of the bone. The level of stiffness of NiTi can be tuned through introducing porosity to match the desired level.In our study, we focus on the design of a patient-specific stiffness-matched fixation hardware as a substitute for currently used fixations (Ti-6Al-4V). The purpose is to minimize the risk of implant failure  and to produce the proposed fixation in a patient-specific manner. The fixation design is based on the exact shape and the required stiffness obtained by the CT-scan data of a dried mandible. Adding a 45.7% porosity to NiTi fixation hardware during 3D printing allows us to decrease the stiffness of fixation plates to stiffness of the surrounding bone (estimated from CT data). In this work, we used finite element analysis to evaluate the effect of using this reduced stiffness for the post-surgical process. Based on this, two models of MRS were simulated in different loading scenarios. Our finite element results indicate a higher level of contact pressure (177%) during initial periods of healing due to pre-tension load in the case of using porous NiTi fixation plates. Such improvements can increase the chance of vascularization during the healing period ,35. AlsoThese porous fixation plates may be fabricated using additive manufacturing (3D printing). For showing a patient-specific design for the fixation plates, a dried cadaver mandible was used. We successfully applied the design process for a case of reconstruction surgery on a dried cadaver mandible (patient-specific study). After the design stage, using Selective Laser Melting (SLM) we could fabricate these patient-specific fixation plates and assemble them on the dried mandible. These 3D printed, porous, NiTi fixation plates not only provide the required level of stiffness (12 GPa) and minimize the stress shielding probability, they also ensure hardware well-fitting that can be quickly placed, thereby reducing operation room time.We have compared the use of NiTi and Ti-6Al-4V as the material for a patient-specific fixation plate. To this end, we created two different porous specimens with the same equivalent Young’s modulus of 12 GPa (estimated from CT scan data) using Ti-6Al-4V and Ni-rich NiTi. The required level porosity for the two specimens was different, 45.7% for the NiTi and 78.3% for the Ti-6Al-4V. Then we applied two percent equivalent strain (the recoverable strain by the cortical bone ,15,16,73


In this research, stabilisation of oil-in-water emulsions with non-chemically modified gelatinised starch is presented. Thus far only octenyl succinic anhydride (OSA) modified gelatinised starch has been known to adsorb at emulsion droplet interfaces, acting as emulsifiers. Screening a range of commercially available food starches revealed that a non-waxy rice starch, a waxy rice starch and the waxy maize starch PRIMA600 showed oil-in-water emulsifying ability following gelatinisation. The microstructure of emulsions formulated with 20% oil and 1% starch was stable for at least 3 months. Thermal, crystallinity and molecular property analyses as well as amylose and protein content revealed no obvious link to this property. Nevertheless, this research has provided the food industry with exciting results for the formulation of clean label emulsions. Moreover, it presents a concept for oral release food emulsions with destabilisation via salivary amylase digestion of the stabilising starch emulsifier.  •Non-chemically modified gelatinised starches screened for emulsifier functionality.•High shear overhead processing of oil-in-water emulsions.•A waxy & non-waxy rice & waxy maize starch adsorbed at oil droplets, others did not.•Not related to thermal, crystallinity & molecular properties, amylose, protein.•Functionality as clean label emulsifier clearly demonstrated. They are two-phase liquid dispersions where one phase is present in the form of droplets in another immiscible phase. With the exception of microemulsions they are thermodynamically unstable. Therefore, in practice emulsion microstructures are kinetically trapped through the use of emulsifiers and often also the addition of viscosity enhancers to the continuous emulsion phase. Emulsifiers adsorb at the interface between the two immiscible liquid phases imparting electrostatic or steric barriers between neighbouring and colliding droplets thus preventing coalescence. Adsorption occurs during processing, trapping process induced microstructures if the concentration of added emulsifier is high enough compared to the total interfacial area of the emulsion. Otherwise, coalescence will occur until surface saturation traps the evolving microstructure. Viscosifiers aide the prevention of coalescence by slowing the creaming or sedimentation of individual droplets thus close proximity of droplets which may lead to coalescence.The research reported here is concerned with emulsifiers suitable to stabilise oil-in-water (o/w) food emulsion interfaces during processing, imparting microstructure stability over storage, but destabilising the emulsion microstructure during oral processing to release in the oil phase entrapped salt or sugar solution. The microstructure of such systems is known as water-in-oil-in-water emulsions generally referred to as multiple, duplex or complex emulsions. The release of the tastant solution from the internal water phase to the vicinity of the taste receptors will impart a higher perceived tastant intensity compared to an emulsion where all of the tastant is present in the external emulsion phase, which we have reported for the case of saltiness enhancement . The emuClearly, native starch with no sites of the molecule rendered inaccessible to amylase would provide the fastest digestion kinetics . It is t22.12.1.12.1.2Iodine for the amylose assay and sodium azide as antimicrobial agent were purchased from chemicals suppliers . Sunflower oil, serving as the dispersed emulsion phase, was purchased in a local supermarket . Dimethyl sulfoxide (DMSO) as solvent for starch, Rhodamine B to fluorescently stain starch and Nile Red to fluorescently stain oil were also purchased from a chemicals supplier . Fast Green to fluorescently stain protein was purchase from Merck All chemicals were used as received. Water used in this research was milli-Q water.2.22.2.1The moisture content of the starches was determined using a moisture analyser . 0.57–0.62 g of starch were weighed into an aluminium pan that was then placed into the equipment fitted with a thermogravimetric balance. The sample was dried at 120 °C and the results are presented in 2.2.2The starches were analysed for their amylose content using a colorimetric assay following a modified published protocol . 5 mg of2.2.3Protein content was estimated by measuring nitrogen by standard combustion method . 5–6 mg 2.2.4onset), peak temperature (Tpeak), endset temperature (Tendset) and enthalpy of gelatinisation (ΔH (J/g) utilising the equipment's software (STARe). Samples were analysed in duplicate and results (mean and standard deviation) have been included in The thermal properties of the starches in excess water were analysed with differential scanning calorimetry . Each starch was mixed at a ratio of 1:3 with water for 20 h. Approximately 5 mg of starch dispersion were then weighed into stainless steel pans . The pans were hermetically sealed, placed into the calorimeter and temperature ramped up from 5 °C to 90 °C at a rate 10 °C/min. An empty pan was used as reference. The data traces were analysed for onset temperature  equipped with a copper tube operating at 40 kV and 40 mA produced Cu Kα radiation of 0.154 nm wave length with a rotation speed of 60 rpm. The samples were examined at 20 °C over the angular range of 3θ to 38θ.2.2.6Molecular weight properties of the starches were assessed by means of their sedimentation coefficient distribution , popular−13 s. Based on previous amylopectin in water hydrodynamic data . Measurement conditions were 25 000 rpm and 20.0 °C. Data were analysed using the least squares ls-g*(s) method with SEDFIT . All disThis formula derives from the power law or scaling relationa is the power law coefficient from the corresponding equation to (1) linking intrinsic viscosity with molecular weight. a has been found to be 0.31 , followed by dilution with water prior to analysis.2.32.3.1endset for all starches with the exception of the high amylose maize starch (HylonVII), see Starches were screened for emulsifying ability by processing o/w emulsions containing 20% oil and 1% starch . The remainder was water containing 0.02% sodium azide to prevent microbial spoilage of the emulsions. In each case 100 g of emulsion were prepared through processing with a high shear overhead mixer  applying a two-step process; starch gelatinisation followed by emulsifying the oil. Initially, the appropriate amount of starch and water were weighed into a tall form 200 mL glass beaker and processed at 8000 rpm for 5 min. During this process the temperature increased from ambient (around 20 °C) to between 60 and 70 °C. Afterwards, without interruption, 20 g of oil were added followed by further mixing at 8000 rpm for 5 min during which temperature increased further. Final temperatures varied between 75 and 85 °C. The temperature profile the starches experienced during sample preparation is illustrated exemplary in Following processing the emulsions were transferred into 50 mL tall capped glass vials and stored at 20 °C for visual assessment of creaming and coalescence. A yellowish appearance of the cream layer, clearly visible large oil droplets, i.e. with a diameter of plus 1 mm, and/or the formation of a bulk oil layer on top of the cream phase were taken as signs of coalescence. Starches applied in emulsions showing coalescence were identified as non-emulsifying. Non-emulsifying starches were then re-evaluated at 2, 3 and 4% of starch to validate their classification as non-emulsifying. Emulsifying starches were also applied at these higher concentrations, with one exception mentioned in the results section, to assess impact on droplet size and other emulsion properties as described in the following.2.3.2To validate droplet size distribution data and to assess droplet flocculation, emulsion microstructure was visualised using bright field illumination on a digital microscope . Samples were prepared by placing a drop of emulsion onto a glass slide followed by carefully sliding over a glass cover slip.Confocal laser scanning microscopy was applied to validate the presence of starch and lack of the presence of protein at the o/w interface. A drop of emulsion was placed onto a glass cover slide, carefully mixed at a ratio of 1:2 with Rhodamine B solution , Fast Green solution  and Nile Red  using a pipette tip followed by sliding over a glass cover slip. Nile Red was used to stain lipid, Fast Green to stain protein and Rhodamine B to stain starch. The sample was then imaged with a confocal laser scanning microscope equipped with an Argon laser (488 nm excitation wavelength) and HeNe laser . A band pass filter between 580 and 620 nm was selected for the detection of Nile Red; when excited at 488 nm, and 651–710 nm for the detection of Fast Green; when excited at 633 nm. A band pass filter for Rhodamine B was selected between 550 and 580 nm and excited at 561 nm. Images were acquired using an ×60 objective.2.3.3Droplet size distributions of the o/w emulsions were acquired using a low angle diffraction particle analyser  fitted with a liquid dispersion cell  containing water. Appropriate amount of emulsion, as indicated by the equipment software, was added to the water before measurement commenced. Three diffraction patterns were averaged and analysed by the equipment software based on the refractive index of 1.333 for water, as the continuous dispersion phase, and 1.54 for the material adsorbed at the surface of the oil droplets following a published method . Results2.3.43 volume into the oil phase contained in a quartz glass cuvette. Droplet volume was held constant during the measurement and all measurements were conducted at 20 °C.A drop shape tensiometer  was used to assess interfacial tension at the oil/water interface where the water phase was pure water, water containing 4% OSA starch or 4% gelatinised waxy rice starch. The starch dispersion was obtained by mixing starch with water at 8000 rpm for 10min using the same high shear overhead mixer as for emulsion processing. The sample was then allowed to cool to room temperature (approx. 20 °C) by placing onto the lab bench before being applied to the syringe of the drop shape tensiometer. This syringe was fitted with a straight stainless steel capillary with an outer diameter of 2 mm and used to dose a pendant water drop of 25 mmAcquisition of interfacial tension data at 20 °C was representative of our observations with regard to the stabilisation of emulsions at much higher temperature (75–85 °C). Those starches stabilising emulsions during the high temperature process successfully also stabilised emulsions processed after cooling the gelatinised starch to 20 °C followed by emulsifying in the oil with the same high shear process but maintaining this lower temperature through cooling with ice.33.13.1.1All of the starches shown in 3.1.2Stable emulsions, i.e. those prepared with the non-waxy rice starch, the waxy maize starch PRIMA600, the waxy rice starch and the OSA starch were analysed for microstructure and droplet size distribution. With regard to the acquisition of droplet size distributions on the emulsions the non-waxy rice starch stabilised emulsion was excluded from this assay. Sampling the emulsion gel into the liquid dispersion cell proved challenging and results would have been representative of large flocs of droplets or small fragments of emulsion gel instead of primary droplet size. The droplet size distributions of the remaining emulsion systems are presented in 3.1.3To develop an understanding of the origin of the emulsifying ability of waxy rice starch the interfacial tension at the o/w interface was measured utilising the pendant drop technique. As described in the methods section preparation of the waxy rice starch for this measurement included high temperature processing as in the case of emulsion processing followed by cooling to room temperature (approx. 20 °C). It was validated that this starch dispersion would stabilise o/w emulsions through high shear processing at 20 °C before interfacial tension measurement at 20 °C. In the absence of any starch interfacial tension asymptotically reached a value of 26.5 ± 0.4 mN/m after a slightly decreasing trend for the first 120 s due to the presence of naturally present surface-active molecules in sunflower oil. The data traces overlapped with the data traces acquired in presence of gelatinised waxy rice starch in the aqueous phase; in this case the averaged value after levelling off was 26.8 ± 1.3 mN/m. However, in presence of OSA starch in the aqueous phase, following an initial longer decrease, the interfacial tension levelled off at 18.2 ± 0.1 mN/m. OSA starch has been created to be interfacially active and a decrease in interfacial tension was expected. However, it appears that gelatinised waxy rice starch did not adsorb at the interface during the interfacial tension analysis leading to the conclusion that it is not interfacially active. As this starch was still stabilising o/w emulsions it was hypothesised that, during high shear processing, molecules or molecular aggregates of starch would contact the o/w interface and then adhere to it imparting the observed emulsion stability. The presence of starch at the o/w interface was validated by confocal laser scanning microscopy; see 3.23.2.1Thermograms of all of the starches acquired in excess water showed a single endothermic peak. The onset, peak and endset temperature of the transition from granular to gelatinised starch and enthalpy of this phase transition were reported in The thermograms acquired on the processed emulsions stabilised with the non-waxy rice starch, the waxy rice starch and the waxy maize starch PRIMA600, lacked an endothermic peak indicative of the fully gelatinised state of the starch in the system.3.2.2The crystallinity pattern of each starch was analysed with X-ray diffraction to inspect whether amylose-lipid complexes ought to be considered in explaining the emulsifying ability of some but not other starches included in this study. Amylose-lipid complexes can be naturally present in native starches  and form3.2.3The sedimentation coefficient distributions for all starches, as they were present in aqueous dispersion prior to the addition of oil in the emulsification process are presented in It is worth stressing here that the sedimentation coefficient distribution data are representative of the starch processed in aqueous environment exactly the way as the emulsions in terms of shear, time and temperature. Water is not a good solvent for starch and therefore the data do not relate to individual starch polymers but to aggregates of starch polymers as confirmed by the multimodal nature of the distributions.s of ∼400 S. The high amylose maize starch Hylon VII, the potato starch and the regular rice starch, all three not showing emulsifying functionality, had narrower sedimentation coefficient distributions at the lower end of the Svedberg unit range (<∼200 S), typically with fewer peaks. The pea starch, also not emulsifying, had a high peak in the lower Svedberg unit domain with a broad low intensity population covering the Svedberg unit range up to 500 S with a small number of peaks that were not isolated from each other. The wheat starch, again not emulsifying, showed a similar pattern although with a larger number of isolated peaks or populations in the mid to high Svedberg unit domain. The sedimentation coefficient distribution of the tapioca starch, which stabilised the o/w emulsion processed in this study when applied at 4%, appeared similar to that of pea starch but with larger populations at the higher Svedberg unit domain.The results presented in Since it was already known which starches showed emulsifying ability the sedimentation coefficient distribution data were assessed if a certain fraction of starch polymers or polymer aggregates at the higher size  range was required for starches to show this functionality. The distributions are shown in 4It can be concluded that at least three non-chemically modified starches exist that will stabilise oil-in-water emulsions processed under high shear as long as the starches gelatinise during processing. This ability to act as emulsifier through interfacial adsorption during processing was not related to amylose content or crystallinity pattern. Characterising the molecular characteristics of the starches and the fraction that remained in the emulsion serum phase with the method of analytical ultracentrifugation (AUC) also failed to increase understanding of criteria for emulsifying ability. Nevertheless, this research has, to the best of our knowledge, for the first time shown that waxy rice starch, non-waxy rice starch and waxy maize starch PRIMA600 act as emulsifiers. So, there are natural or clean label starches, not only alternative to OSA starch but also to other synthetic food emulsifiers, that can be applied for the interfacial stabilisation of emulsion based foods.


Traditionally, every year, the International Brazilian Journal of Urology evaluates its reviewers and selects the most efficient along the previous year, Evaluation is based on the number of performed reviews, time for conclusion and quality of work .In 2017, the five most efficient reviewers were:Fabio Vicentini , Eduardo Kaiser Ururahy Nunes Fonseca , Victor Srougi , Elcio Silva  and Kemal Sarica .We would like to thank them and all others reviewers, that are the reason for the existence of a peer-review scientific journal.Int Braz J Urol is an open-access magazine, integrally supported by the Brazilian Urological Society, that does not charge for the submission of papers. Also, it is included in the Urology Green List , that reIn 2017, our Journal received 603 papers, and 28% were accepted. Medium time for first decision correspondence was 44 days, and more 42 days for publishing as Ahead of Print. Papers were sent from 42 countries, from 635 different institutions and 2254 authors; we used 1135 reviewers. The 5-year impact factor of IBJU in 2017 was 1.097.