One is a shortage of intra-abdominal space for the liver graft, and the other is the size discrepancy between the anteroposterior dimensions of the liver graft and the lower right hemithorax of the recipient. The former could be treated using delayed fascial closure or mesh closure, but the latter may only be treated by reduction of the right liver graft to increase space. Given that split liver transplantation has strict requirements regarding donor and recipient selections, reduced-size liver transplantation, in most cases, may be the only solution. However, surgical strategies for the reduction of the right liver graft for adult liver transplantations are relatively unfamiliar. Herein, we introduce a novel strategy of HuaXi-  Large-for-size syndrome (LFSS) usually occurs in paediatric liver transplantation (LT) due to the implantation of an excessively large liver graft into a small recipient cavity, resulting in poor graft or recipient outcomes., 2 Howevex vivo right posterior sectionectomy (eRPS) while preserving the right hepatic vein (RHV) in the graft to prevent LFSS and propose its initial indications.There are two causes of graft compression in LFSS. One is a shortage of intra-abdominal space for the graft, and the other is the size discrepancy between the anteroposterior dimensions of the graft and the lower right hemithorax of the recipient. The former could be treated using delayed fascial closure or mesh closure; however, the latter may only be treated by reduction of the right liver graft to increase space. Given that split liver transplantation (SLT) has strict requirements for donor and recipient selections, reduced-It is dangerous for donors to undergo computed tomography (CT) examinations during organ maintenance in the intensive care unit (ICU), although CT is the most accurate method to measure the graft’s right anteroposterior (RAP) vertical distance and the largest horizontal distance. Hence, in our centre, we do not perform CT imaging on donors to ensure the safety of donors during organ maintenance in the ICU. eRPS was performed in five grafts between January 2019 and November 2020.Regarding the recipients, we defined the longest RAP vertical distance between the anterior and posterior parts of the ribs at the lower extremity of the xiphoid process on a CT scan . Both grAll organs were donated after death, and no organs were obtained from executed prisoners. eRPS was performed on the back table. The primary cutting plane was designed according to the right side of the RHV root into the suprahepatic inferior vena cava (IVC), right edge of the retrohepatic IVC, and Rouviere’s sulcus . ParenchIn this study, HuaXi-eRPS was performed in five grafts. The five donors did not meet the criteria for split candidates utilised by UNOS; thus, SThe 30-days mortality was zero. Postoperative complications occurred in two patients (40%); however, complications higher than those in Clavien-Dindo grade II were notThe morphology of the right upper abdominal cavity may differ among individuals. To date, four formulas have been proposed to predict the occurrence of LFSS., 12-14 H However, it is very unlikely to solve some mismatch issues because compression, due to the ribs, mainly applies to the right liver. Right hemihepatectomy has been proposed as an alternative method, but the residual left liver may be insufficient for some recipients.(in vivo method, the HuaXi-eRPS used in our series could be a unique method with the following advantages. First, the graft weight can be accurately measured on the back table to provide a precise parameter for determining the feasibility of eRPS. Second, because the ex vivo graft can be rotated 360-degree, it is easy and simple to perform eRPS using the cranial approach to the RHV. Although the demarcated area for the right posterior sector cannot be displayed easily after ligating the right posterior Glisson’s sheath as an in situ graft, the main purpose of eRPS is to overcome size mismatch. It is not necessary to perform a precise anatomic right posterior sectionectomy, as required for hepatic malignancy. Third, eRPS in the graft before implantation is beneficial to reduce the difficulty of implantation and shorten the period for the anhepatic phase. In addition, compared to the whole right lobe, which accounts for 60–75% of the total liver volume, eRPS can ensure both the integrity of outflow and adequate residual graft volume to avoid SFSS while avoiding rib compression.Compared to paediatric RSLT,, 16 the cipients. ComparedThe present study had some limitations. GW/RAP combined with GRWR, as a new “LFSS predictor,” is a preliminary formula whose optimal cutoff value or predictive validity still requires further confirmation by a well-designed trial with a large sample size. However, this is the first study to propose the initial indications for HuaXi-eRPS in grafts, and its initial outcomes in our five adult series are safe and encouraging, especially in decreasing the difficulty of implantation, avoiding delayed fascial closure, shortening ICU stay, and reducing posttransplant complications.In conclusion, this study described a novel and feasible surgical strategy for preventing posttransplant LFSS, especially for the size discrepancy between the anteroposterior dimensions of the liver graft and the lower right hemithorax of the recipient.This study describes a novel and feasible surgical strategy for preventing posttransplant large-for-size syndrome, especially for the size discrepancy between the anteroposterior dimensions of the liver graft and the lower right hemithorax of the recipient.


Vibrio cholerae, and young children have poor immunological responses to bacterial polysaccharides, which are T cell independent antigens. To overcome this, we have developed a cholera conjugate vaccine (CCV) containing the OSP of V. cholerae O1, the main cause of endemic and epidemic cholera. Here, we describe production of CCV through a scalable manufacturing process and preclinical evaluation of immunogenicity in the presence and absence of aluminum phosphate  as an adjuvant. The vaccine displays V. cholerae O1 Inaba OSP in sun-burst display via single point attachment of core oligosaccharide to a recombinant tetanus toxoid heavy chain fragment (rTTHc). Two different pilot-scale production batches of non-GMP CCV were manufactured and characterized in terms of physico-chemical properties and immunogenicity. In preclinical testing, the vaccine induced OSP- and lipopolysaccharide (LPS)-specific IgG and IgM responses, vibriocidal responses, memory B cell responses, and protection in a V. cholerae O1 challenge model. The addition of alum to the administered vaccine increased OSP-specific immune responses. These results support evaluation of CCV in humans.There is a need to develop cholera vaccines that are protective in young children under 5 years of age, which induce long-term immunity, and which can be incorporated into the Expanded Programme of Immunization (EPI) in cholera-endemic countries. The degree of protection afforded by currently available oral cholera vaccines (OCV) to young children is significantly lower than that induced by vaccination of older vaccine recipients. Immune responses that protect against cholera target the O-specific polysaccharide (OSP) of  For immunologic assays, OSP was conjugated to bovine serum albumin (BSA Sigma) as previously described V. cholerae O1 PIC018 as previously described Production of CCV was advanced from previous bench-top non-scalable production 2.2Vibrio cholerae strains, involved developing a V. cholerae O1 El Tor PIC018 seed-inoculum in shake-flasks in semi-defined medium containing salts, yeast extract , amino acids  and antibiotic , followed by inoculation of seed culture into a production fermenter with the same medium. A feed strategy was used to increase cell density using medium containing sugar, salts, yeast extract, amino acids and antibiotic. Following V. cholerae O1 Inaba fermentation, acid-hydrolysis was performed after cell concentration and washing by microfiltration/diafiltration  in tangential flow filtration (TFF) mode; and cellular debris was removed by centrifugation  and 0.2 µm filtration. OSP purification then involved application of 30 kDa cut-off membrane cassettes  in TFF mode; and permeate was recovered and processed again through 5 kDa cut-off membrane cassettes  in TFF mode, with the retentate from the latter step concentrated and diafiltered into final buffer followed by sterile-filtration using a 0.2 µm filter. Final purified OSP was analyzed for purity via SE-HPLC , polysaccharide content via resorcinol assay, and protein content via BCA (bicinchoninic acid) assay.Production of Inaba O-specific polysaccharide (OSP), as an integral component of lipopolysaccharide (LPS) in Escherichia coli-rTTHc strain seed-inoculum in shaker-flasks containing APS-LB (Alternate Protein Source-Luria-Bertani) medium  and an antibiotic, and inoculation of seed culture into a production fermenter with the same medium. A feed strategy was used to increase cell density using medium containing sugar, yeast extract and peptone. Target protein induction was done using isopropyl β-D-1-thiogalactopyranoside  and galactose  combination strategy using an auto-induction mechanism. The fermenter culture was harvested by centrifugation  once the depletion of feed nutrient was observed. The harvested cell paste was frozen at –70 ± 10 °C until used for rTTHc isolation and purification. rTTHc was purified from supernatant obtained after lysis of cell paste. rTTHc purification involved Ion-Exchange  and Hydrophobic Interaction  chromatography steps, as well as a TFF step . Final purified rTTHc was analyzed for purity via SEC-HPLC , sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis, and protein content via BCA assay.Production of rTTHc involved developing a recombinant For making CCV conjugate , purifieTo assess reproducibility and production parameters, two batches of CCV were made: EubB2006-1 and EubB2006-2. To produce EubB2006-1, the reaction involved mixing 1.7 g of purified OSP, labelled with DSE and concentrated to ∼ 50 mg/mL, with 0.8 g of the concentrated rTTHc (30 mg/mL) in 50 mM sodium borate buffer (pH 9.8) and made into a final volume of 65 mL. This entailed a starting OSP concentration of 26 mg/mL. To produce EubB2006-2, the reaction involved mixing 1.7 g of the purified OSP, labelled with DSE and concentrated to ∼ 50 mg/mL, with 0.8 g of the concentrated rTTHc (30 mg/mL) in 50 mM sodium borate buffer (pH 9.8) and made into a final volume of 130 mL. This entailed a starting OSP concentration of 13 mg/mL.2.3V. cholerae O1 Inaba serotype (n = 5) versus Ogawa serotype (n = 5). We coated wells (100 ng/well) with either OSP:rTTHc EubB2006-1, OSP:rTTHc EubB2006-2, OSP:BSA, or rTTHc, and assessed immunoreactivity via ELISA using human sera (1:250 dilution) as previously described 2O2 (Sigma), and optical density at 405 nm was determined with a Vmax microplate kinetic reader (Molecular Devices Corp.). Plates were read for 5 min. at 30 s intervals, and the maximum slope for an optical density change of 0.2 U was reported as millioptical density units per minute (mOD/min.) To assess immune-recognition of the OSP display on CCV, we performed antigen-specific ELISAs using pooled sera collected from humans with cholera in Bangladesh, and compared these responses to controls of humans with typhoid fever (n = 5) in Bangladesh. We separately assessed immune responses in humans with cholera caused by 33.1We immunized female Swiss-Webster mice  with 10 µg of polysaccharide per dose of OSP:rTTHc of EubB2006-1 or EubB2006-2. A separate cohort of mice was immunized with EubB2006-1 with aluminum phosphate , mixed in a 1:1 volume per manufacturers’ instruction. Control cohorts included mice vaccinated with aluminum phosphate alone, or Phosphate Buffered Saline (PBS) alone. To assess kinetics of immune responses, mice were vaccinated intramuscularly on days 0, 14, 28, and then again on day 56. We collected blood samples via tail bleeding on days 0, 7, 14, 21, 28, 35, 56, 63 and at sacrifice. Samples were collected, processed, aliquoted, and stored as previously described 3.2We assessed OSP and TT-specific IgG and IgM responses in serum using standard enzyme-linked immunosorbent assay (ELISA) protocols as previously described  3.3V. cholerae O1 El Tor Inaba strain PIC018 as previously described We assessed serum vibriocidal antibody titers against 3.45 splenocytes at the time of sacrifice.Nitrocellulose bottom plates  were coated with OSP:BSA (100 ng/well), or keyhole limpet hemocyanin   or goat anti-mouse IgG (Southern Biotech) 3.5V. cholerae O1 El Tor Inaba strain N16961 as the challenge strain. In brief, we removed 3–5 day-old un-immunized CD-1 suckling mice from dams two hours prior to inoculation. We then administered to pups a 50 μL inoculum comprised of 1x109cfu of V. cholerae O1 El Tor Inaba strain N16961 mixed with a 1:1 dilution of pooled day 63 sera from mice intramuscularly immunized with placebo, OSP:rTTHc, or OSP:rTTHc with aluminum phosphate. Following oral challenge, we kept neonates at 30 °C and monitored animals every 3 h for 36 h, after which surviving animals were euthanized.To assess protection afforded by vaccination, we used the standard cholera neonatal mouse cholera challenge assay, as previously described 3.6V. cholerae O1 were collected from patients at the International Centre for Diarrhoeal Disease Research in Dhaka, Bangladesh . This study was approved by the Ethical Review and Research Review Committees of the icddr,b, and the MassGeneral Brigham (MGB) Institutional Review Board, Boston, Massachusetts, USA. Animal work was performed in accordance with all governmental and institutional requirements. All protocols were reviewed and approved by the Massachusetts General Hospital Subcommittee on Research Animal Care (SRAC).Plasma samples from humans recovering from cholera caused by 3.72) tests. Except for vibriocidal analysis that was one-tailed, all reported P values were two-tailed, with a cutoff of P ≤ 0.05 considered a threshold for statistical significance. We performed statistical analyses using GraphPad Prism 5 We compared data within groups across time points using Wilcoxon Signed-Rank tests, and across groups using Mann-Whitney U tests. We compared response rates using chi-square , compared to mice challenged using sera from mice vaccinated with placebo alone , evaluation of immunoreactivity using serum derived from humans surviving cholera, and preclinical assessment of CCV. Currently available oral cholera vaccines (OCVs) have been a transformational addition to global cholera control programs; however, a in vitro bactericidal assay largely comprised of IgM and IgG responses against V. cholerae OSP V. cholerae pathogen should be considered in the context that there is no convincing evidence that the terminal complement components required to form a membrane attack complex are present within the intestinal lumen in the setting of an intact intestinal epithelium, the fact that IgA does not bind complement via the classical pathway , and that data do not suggest any evidence for actual killing of V. cholerae within the intestines of infected humans or animals who are protected from disease Historically, the vibriocidal response has been used as an imperfect correlate of protection against cholera. Although there is a correlation of vibriocidal titers and protection against cholera, there is no absolute vibriocidal value that predicts protection against cholera V. choleraeV. cholerae motility, and this effect may be mechanistically involved in protection against cholera within the intestinal lumen of infected humans V. cholerae pathogens from reaching their target ecological niche within the intestinal lumen of infected humans and delivering cholera toxin to intestinal epithelial cells may be protective In summary, there is no evidence that bacteriocidal activity is a mechanism of protection against cholera. In comparison, a growing body of evidence suggests that protection against cholera is mediated by OSP-specific antibody responses V. cholerae exist, with serogroups O1 and O139 capable of causing epidemic cholera, and protection against cholera is serogroup-specific V. cholerae is a relatively simple structure, comprised of repetitive (1 → 2)α-linked D-perosamines, which are N-acylated with 3-deoxy-L-glycero-tetronic acid, with a chain length of approximately 7–22 perosaminyl residues. V. cholerae O1 organisms can be classified into two major serotypes, Ogawa and Inaba, based on the presence or absence, respectively, of a methyl group on the upstream terminal 2-OH moiety of OSP V. cholerae LPS. Previous infection with Ogawa provides protection against subsequent Ogawa infection, while previous infection with Inaba provides more complete protection against both subsequent Inaba and Ogawa infection V. cholerae O1 Inaba El Tor PIC018, a well-characterized strain isolated in 2007 from a patient in Bangladesh, as our source strain (GENBANK: https://www.ncbi.nlm.nih.gov/nuccore/NZ_LQHM00000000.1).Over 200 serogroups of V. cholerae directly in biofermenters, retaining intact the OSP-core component, which contains an active glucosamine group. We then use squarate chemistry to directly link V. cholerae OSP via core oligosaccharide to carrier protein. This approach does not require introduction of an unrelated linker, minimizes derivatization steps, simplifies manufacturing, and significantly lowers the cost of production of CCV. It results in conjugates that are not cross-linked, easy to characterize, and display OSP in a sun-burst, single-point-attachment in a manner that recapitulates how OSP is presented on V. cholerae. It should be noted that other Gram-negative bacterial pathogens also contain a single active glucosamine in core oligosaccharide, including Shigella spp., suggesting that our conjugation approach could have platform utility in producing multi-valent enteric vaccines.Our production protocol uses mild hydrolysis of LPS from E. coli for production purposes, and functions very well in conjugation reactions and as a carrier protein We chose a recombinant 52 kDa fragment of tetanus toxoid heavy chain as carrier protein. This protein was optimized for high level expression and purification in V. cholerae O1: Inaba and Ogawa. This is not unexpected since the two serotypes have high homology, differing only in the presence or absence of a methyl group on the terminal saccharide of OSP. Immune responses and protection against cholera is not serotype specific We found that OSP displayed on CCV was recognized by plasma from humans recovering from cholera caused by both of the major serotypes of Salmonella Typhi in the United Kingdom In our preclinical immunogenicity analysis, we found that OSP-specific immune responses were boosted in the presence of aluminum adjuvant. The presence of alum did not markedly increase survival in our protection assay. There was a trend in increase in LPS-specific and rTTHc-specific IgG but not IgM responses in the presence of alum. There was, however, no evident increase in vibriocidal responses, perhaps reflecting the absence of boosting of IgM responses by alum. We did find that vaccination with CCV with or without alum induced IgM responses in addition to IgG responses but did not induce rTTHc-specific IgM responses. This may reflect different immune processing of polysaccharide versus protein antigen. Analysis of CCV showed absence of unconjugated OSP. Interestingly, vaccination with a typhoid conjugate vaccine similarly induced polysaccharide-specific IgM and IgA responses not only in humans vaccinated in a typhoid endemic zone (with possible boosting of pre-existing responses) Haemophilus influenzae b, Streptococcus pneumoniae, and Neissseria meningitidis, not only in preventing invasive disease but also in decreasing mucosal colonization and carriage with serotypes included in the conjugate vaccine, suggesting a mucosal effector function Analysis of previous conjugate vaccines are also consistent with induction of impactful mucosal responses. It has long been recognized that parenteral immunization can boost both mucosal and systemic immune responses that protect against cholera in humans residing in cholera-endemic areas, possibly through boosting of pre-existing immune responses, including IgM and secretory IgA in intestinal secretions, milk and saliva IgA and IgM are actively secreted into the lumen at the intestinal surface, but anti-polysaccharide IgG is also detected in mucosal fluids following parenteral vaccination with conjugate vaccines, even in young infants, but not following vaccination with unconjugated polysaccharide V. cholerae OSP in an immunologically relevant manner, is recognized by convalescent phase sera from both Inaba and Ogawa serotype-associated cholera, and induces protective and durable memory immune responses in pre-clinical evaluation studies. OSP is the target antigen for immune responses that mediate protection against cholera, and polysaccharide-specific immune responses following immunization with conjugate vaccines can be protective at mucosal surfaces. Our approach is an improvement over previous unconjugated parenteral cholera vaccines that were as protective as oral cholera vaccines, but that induced only short-term protection requiring frequent re-vaccination. Our approach also addresses the shortcomings of currently available oral cholera vaccines that suffer from poor protective efficacy in children under 5 years of age, induction of relatively short-term protection, and difficulties in incorporating into EPI schedules. Our data suggest that CCV, following completion of a GLP toxicology study, should be evaluated in a phase 1 trial in humans to evaluate safety and immunogenicity. This and subsequent evaluations should include assessment of immune responses associated with protection against cholera, including OSP-LPS-specific responses in serum, OSP-LPS-specific responses in mucosal samples , and OSP-LPS memory B cell responses.In conclusion, we report here development and scalable production of a cholera conjugate vaccine. We demonstrate that the vaccine displays The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


After an investigation into reader concerns, the Medicine Editorial Office found that passages had been directly plagiarized and/or slightly modified from “Low vision rehabilitation for better quality of life in visually impaired adults”. The categorization of interventions, pooling of data, a flow chart, references, and data in forest plots were also plagiarized.The article “Low vision rehabilitation in improving the quality of life for patients with impaired vision: A systematic review and meta-analysis of 52 randomized clinical trials”,International Journal of Surgery while it was still under consideration by Medicine. International Journal of Surgery withdrew the epublication for similar concerns.In addition to plagiarism concerns, included studies Stroupe 2018, Stelmack 2017, Brody 2006, and Brody 2002 were misrepresented as four separate studies. Stroupe 2018 belongs to the same study as Stelmack 2017, and Brody 2006 is a subsample of Brody 2002. It was also discovered that authors submitted the publication to


JAMA article titled ‘The Complementary Role of Environmental and Security Biological Control Regimes in the 21st Century’, taking into account the international policy developments that have occurred over the past two decades. The paper underscores the utility of the concept of a ‘web of prevention’ for elucidating the need for continuous interaction between the international biosafety and international biosecurity regimes, in order to ensure that the life sciences are used only for peaceful purposes. The terms ‘biosafety’ and ‘biosecurity’ are used to denote the primary purpose of the two regimes: the international biosafety regime seeks to prevent the unintentional  release of pathogens and toxins, including naturally occurring disease, whereas the biosecurity regime seeks to prevent the deliberate release and misuse of pathogens and toxins. The paper concludes by recommending practical steps for strengthening the implementation of all elements of the web of prevention and upholding the norms against the hostile misuse of life sciences.Biological threats are complex and multifaceted, as evidenced by the ongoing COVID-19 pandemic. Their effective prevention and countering require multiple lines of collaborative action and sustained cross-sectorial coordination. This paper reviews the conclusions of Graham Pearson’s 1997  There are 2 driving forces for multilateral biological control regimes: international/national security and environmental protection. With respect to deliberately caused disease, these seemingly disparate forces are mutually reinforcing as demonstrated by simultaneous moves to strengthen the Biological and Toxin Weapons Convention and the entry into force of the Convention on Biological Diversity. Future multilateral biological control regimes based on these developments will aid the security, prosperity, and health of the world community”  ecosystems in which they exist’.  At interBiological Threat Reduction Strategy: Strengthening Global Biological Security which seeks to develop a sustainable and effective protection against threats from deliberate and accidental releases of animal, including zoonotic pathogens by strengthening existing systems for surveillance, early detection, and rapid response and fostering scientific networks for promoting biosafety and biosecurity [In 2015, the OIE published security . To senssecurity . The OIEsecurity .FAO Biosecurity Toolkit that was published in 2007 promotes an integrated approach to the management of biological risks related to food safety, zoonoses, the introduction of animal and plant diseases and pests, the introduction and release of LMOs and their products, and the introduction and management of invasive alien species [The  species . The TooGuidance on Risk Assessment of Living Modified Organisms and Monitoring in the Context of Risk Assessment developed within the framework of the Cartagena Protocol defines a five-step roadmap for conducting a risk assessment of LMOs which takes into account risks to human health [Biodiversity plays a critical role in ensuring the health of natural ecosystems and its preservation is indispensable to the implementation of a ‘one health’ approach for realising health security. Given the lasting impact that novel life sciences advances can have on natural ecosystems, it is essential that the application of such advances is carried out in ways that ensure environmental integrity and protect human health. The 1993 Convention on Biological Diversity (CBD) acknowledges the adverse effects that LMOs may have on the conservation and sustainable use of biological diversity and its 2003 Cartagena Protocol on Biosafety sets out specific provisions for assessing and managing the risks related to handling, transfer, and use of LMOs . The Guin health . The GuiIn line with its mandate to monitor and assess the impact of novel biotechnology advances, the CBD Subsidiary Body on Scientific, Technical and Technological Advice (SBSTTA) has recommended the establishment of an Ad Hoc Technical Expert Group on Synthetic Biology which seeks to contribute to the harmonisation of the regulation of organisms, components, or products derived from synthetic biology and identify risks and benefits of the use of synthetic biology techniques to the conservation and sustainable use of biodiversity and related human health and socioeconomic impacts . Key issSafety procedures and practices for handling, shipping, transfer, and transport of biological agents and toxins are intended to prevent laboratory acquired infections and ensure that such materials are not accidentally released in the environment. The International Organisation for Standardisation (ISO) has recently published an ISO Management System Standard titled ISO 35001 on Biorisk Management for Laboratories and other Related Organisations . ISO 350WHO Laboratory Biosafety Manual – Fourth Edition adopts a risk- and evidence-based approach to biosafety that emphasises the importance of a comprehensive safety culture in promoting effective and sustainable laboratory practice and risk management [Both WHO and OIE respectively have developed a body of guidance on laboratory safety . The 202nagement . The Mannagement .JAMA paper:“In considering biological control regimes for the future, it is instructive to look back over the developments in both security and environmental controls.” As noted in the conclusion of 1997 Strengthening the international systems for countering disease threats regardless of their origins and ensuring that effective mechanisms are in place to prevent the hostile misuse of life sciences advances requires the sustained interaction of biosafety and biosecurity regimes. The web of prevention constitutes a useful conceptual tool for developing an understanding of the complementary relationship of these two sets of instruments and devising integrated policy strategies for promoting the effective national implementation of their provisions. Attention needs to be given to the following steps and measures for strengthening the implementation of all elements of the web of prevention:An essential element in the process of harmonising the international biosafety and biosecurity regimes is ensuring that States recognise and respect their legally binding obligations. As of 2021, key international biosafety (e.g. Cartagena Protocol on Biosafety) and biosecurity  instruments have not been ratified by all States. By joining relevant international agreements, States develop an appreciation of the complementary role of these agreements and are committed to take effective steps for their national implementation. At the same time, international legal frameworks provide a normative and procedural context for dealing with instances of non-compliance. As such, they help reassert the importance of international rules by drawing attention to the reputational, economic, and political costs that States could incur if they fail to abide by these rules.It is important that the outcomes achieved in one area of biological risk management are shared promptly into the proceedings of other international agreements, in order to ensure cross-fertilisation and effective data sharing. For instance, those working in the field of disarmament need to be kept up-to-date with international policy developments in the field of health, responsible laboratory practice, and LMOs risk assessment, and vice versa.Biological risk management is complex and multifaceted, and hence it is important that the existing processes for international multilateral negotiations are supplemented with opportunities for a broad stakeholder engagement. Semi-formal or informal forums and platforms  can bring together both government and civil society representatives, including industry and academia, in order to foster the development of innovative solutions and facilitate the transfer of best practices and lessons learned . As suchInternational law defines the legal responsibilities of States in their conduct with each other, and their treatment of individuals within State boundaries . With re


Chasmistes liorus, Jordan, 1878) and Utah sucker  has been a matter of controversy since the mid 1900s. Chasmistes liorus is endemic to Utah Lake, UT and has a subterminal mouth adapted for pelagic feeding. Catostomus ardens is widely distributed throughout the Bonneville Basin and Upper Snake River Basin and has a ventral mouth adapted for benthic feeding. Chasmistes has been recognized as a separate ancient genus. Despite being morphologically distinct, no study has successfully identified residual genetic markers that separate these species. Of these studies, several have used a subset of mitochondrial genes, but no study has analyzed the complete mitochondrial genomes (mitogenomes) of these suckers (Pisces: Catostomidae). To further explore the genetic relationships between these species, we report the complete mitogenomes of Chasmistes liorus and Catostomus ardens. DNA was sequenced using an Illumina HiSeq 2500 system and mitogenomes were assembled and annotated using Geneious v. 2021.2 and MitoAnnotator, respectively. The mitogenomes of Chasmistes liorus and Catostomus ardens are both 16,623 bp and are ∼0.072% divergent. We examine the phylogenetic relationship between Chasmistes liorus and Catostomus ardens using 33 mitogenomes, representing 16 species, from Catostomidae. Our data suggest that Chasmistes liorus is sister to Catostomus ardens. Additional samples from multiple localities and/or cohorts of these species will allow us to better resolve the complicated phylogenetic relationships between these species.The relationship between June sucker ( Chasmistes liorus, Jordan, 1878), is a member of Chasmistes, an ancient genus (four species) of fish endemic to plankton-rich, terminal lakes in western North America. Chasmistes have subterminal mouths adapted for pelagic feeding. Utah sucker , is a member of Catostomus, a genus (26 species) that has ventral mouths adapted for benthic feeding  found in North America and Asia  and benthivorous form (Catostomus ardens). This hypothesis suggests that the presence of intermediates could represent a convergence toward, or divergence away from a June sucker-like morphology, dependent on environmental conditions  evidence of incipient speciation with only a few differences due to limited time of divergence  evidence of a recent hybridization event followed by asymmetric replacement of jerry.johnson@byu.edu) under accession numbers 318670 and 318683.We collected larval June sucker (brood stock population) and Utah sucker  in 2014 from the State of Utah’s Fisheries Experiment Station . We euthanized specimens with tricaine methanesulfonate  and then stored the specimens in RNAlater  at −20 °C . We extracted DNA using the whole specimen and sequenced our samples using an Illumina HiSeq 2500 system  at Brigham Young University’s DNA Sequencing Center . The DNA samples were deposited in the Life Science Museum, Brigham Young University  as a reference. We acquired 31 mitogenomes (14 species) representing all available mitogenomes for Catostomidae from GenBank. We generated multiple sequence alignments separately for each of the 13 protein-coding genes using MAFFT v. 7.475 . Our IQ-tree run used ModelFinder  and Catostomus ardens (MZ907582) were both 16,623 bp, containing 13 protein coding genes, 22 tRNAs, two rRNAs, and the control region. Consistent with previous studies (Mock et al. Chasmistes liorus and Catostomus ardens were very similar. Their complete mitogenomes were ∼0.072% divergent, with all differences occurring in the protein-coding genes. In comparison, the complete mitogenomes of the putative sister taxa Catostomus latipinnis and Xyrauchen texanus were 4.394% divergent.The complete mitogenomes of Chasmistes liorus is sister to Catostomus ardens (Chasmistes and Catostomus, we cannot reject either the incipient speciation or the asymmetric hybridization hypotheses. In addition, the placement of other members of Catostomus is consistent with previous studies that failed to resolve Catostomus as monophyletic (Bagley et al. In our within-taxon-limited phylogeny, s ardens . This is


Hydrolysis of starches with acid decreases swelling power while hydroxypropylation increases the swelling power. The results also showed lower gelatinization  and pasting parameters  for the dual modified starches than other treatments. In summary, this study showed that dually modified tapioca starch has potential application in dip molding and coating.The aim of this study was to investigate the effects of dual modification on the functional, microstructural, and thermal properties of tapioca starch. Tapioca starch was first hydrolyzed by 0.14 M HCl for 0, 6, 12, 18, and 24 hr and then hydroxypropylated by adding 0%, 10%, 20%, and 30% (v/w) propylene oxide. The degree of hydroxypropylation, solubility, water absorption, rheological, thermal, and microstructure characterization of dually modified tapioca starch was determined. Hydroxypropylation did not cause any considerable changes in the starch granular size and shape of tapioca starch. Acid hydrolysis disrupts the starch granules, and the starches with smaller sizes were produced. The degree of molar substitution (DS) of dual modified starches ranged from 0.118 to 0.270. The dual modified starches significantly had higher solubility than native starch ( This study aimed to investigate the effects of dual modification on the functional properties of tapioca starch.Acid hydrolysis disrupts the starch granules, and the starches with smaller sizes were produced.This study showed that dually modified tapioca starch has potential application in dip molding and coating. Bhd. . All chemicals used in this study were of analytical grade.2.2g. The resulting suspension was placed at 50°C for different periods of time  to prepare starch with different molecular weights. The resulting suspension was stirred in a rotary incubator  at 200 rpm to prevent the formation of sediment. After certain periods of time , the starch solutions were neutralized with sodium hydroxide (1% NaOH) until their pH reached 5.5. The samples were washed with distilled water (twice their volume), and then, they were filtered through filter paper (Whatman No. 4). The starch samples were placed in an oven  at 40°C overnight to dry completely  was performed first, followed by hydroxypropylation (HP). For acid hydrolysis of tapioca starch, about 40% of starch solution was prepared by adding 400 g of starch to a solution of 0.14 N hydrochloric acid at 50°C, and the final weight was 1,000 To prepare hydroxypropylated tapioca starch, 20% w/v sodium sulfate was added to the 20% w/v hydrolyzed starch solution and stirred. Using 5% sodium hydroxide, the pH of the solution was adjusted above 10.5. Propylene oxide was added as an etherifying agent in different proportions ). The samples were then capped and stirred at 20–25°C for 30 min. The resulting solution was then stirred in a rotary incubator at 200 rpm to prevent the formation of sediment. After that, using 10% hydrochloric acid, the pH of the suspension was reduced to 5.5. Samples were immediately washed with distilled water until their sulfate content was negative. By the time the moisture content reached about 10%, the samples were dried in an oven at 40°C. After grinding the starch samples, they were passed through a sieve with a size of 250 µm   was used to determining the appearance of starch granules. First, a very small amount of the sample (about 1 g) was glued to a circular metal base made of aluminum, and the surface of the sample was covered with a layer of gold by passing the sputter coater device through a stream of argon gas. The samples were then examined under a microscope with an electric potential of 15 kV  was added to the tube and stirred well. It was then placed in a water bath at 25°C for 1 hr. Finally, with concentrated sulphuric acid, it reached a volume of 25 ml. The resulting solution was transferred to a spectrophotometer  cell, and its absorbance was read at 590 nm. MS of starch was calculated as follow:w is the equivalent hydroxypropyl in 100 g of starch, and M is the molecular weight of C3H6O  at 700 To determine the water uptake of starches, the centrifuge tubes used in the solubility test were weighed along with the sediments deposited in them after the supernatant was removed, and the amount of water uptake was calculated using the following equation  . 4 g of starch sample was added to 25 g of distilled water. The desired speed was 960 rpm in the beginning and 160 rpm at the resting stage of the sample. In this experiment, the starch was first kept at 50°C for 1 min and then rapidly increased to 95°C (at a speed of 14°C per minute) and remained at this temperature for about 3.3 min. It was then cooled to 50°C for 4 min and finally kept at this temperature for 2 min. The pasting properties studied in this study included pasting temperature (°C), time to reach peak viscosity (min), peak viscosity (cP), and final viscosity (cP) , peak temperature (Tp), ending temperature (Te), and gelatinization enthalpy (ΔH), were determined using a differential scanning calorimeter (DSC) . For this purpose, the suspension of starch in distilled water in a ratio of 3:1 (w/w) was weighed in a high‐pressure resistance steel container for the device and kept at room temperature for one day to reach equilibrium. Then, the container containing the sample was placed inside the DSC device, and while heating the sample from 10 to 115°C, the sample was scanned. An empty container of the device was used as a control. The thermal properties of starch samples were calculated and reported from the DSC curves  and Duncan multiply range test to identify the significant difference between samples at 33.1Scanning electron microscopy images of native tapioca starch and starch samples modified by acid hydrolysis, hydroxypropylation, and dual modification are shown in Figure Saberi et al.  similarl3.2p < .05). However, in each of the constant ratios of propylene oxide, increasing the time of acid hydrolysis did not show a significant effect on the MS value of the modified starch samples.In this study, the reaction efficiency of hydroxypropylation and acid hydrolysis was investigated by the degree of molar substitution of hydroxyl groups inside the starch granules. MS involves measuring the average number of hydroxyl groups per unit of anhydroglucose produced by substitution groups . Increasing the oxidizing agent ratio from 10% to 30% and hydrolysis time from 6 to 24 hr also caused a significant increase in water solubility of starches (p < .05). The highest solubility was obtained in the dually modified starch at the highest oxidizing agent ratio and the highest acid hydrolysis time (HP30AH24) (99.49%). According to the results of the microscopic structure of tapioca starches, the increase in solubility of samples due to acid hydrolysis can be interpreted. As the SEM images showed, acid hydrolysis to some extent caused the degradation of starch granules, which in turn increased the amount of amylose out of the granules, resulting in increased solubility in water. Hydroxypropylation also increased the solubility of tapioca starch. This increase in the solubility of starches is associated with placing hydroxypropyl groups with –OH groups in the structure of starch. These hydroxyl groups are hydrophilic and increase access to water molecules . However, acid hydrolysis of starch samples showed an opposite effect on water uptake, so that increasing the time of hydrolysis significantly reduced the amount of water uptake of starch samples (p < .05). The lowest water uptake percentage in modified starch was obtained at the highest hydrolysis time (AH6) (0.94%), and the highest amount was for HP30 (20.58%). Reduction of water uptake of starch granules by acid hydrolysis can be attributed to partial degradation of granule structure by acid treatment. Because due to the destruction of starch granules, their water holding capacity is reduced, and as a result, the water uptake is reduced. Increasing the water uptake of hydroxypropylated starches is due to the fact that due to the entry of hydroxypropyl groups into the starch chains, intramolecular and intermolecular hydrogen bonds are destroyed, and the water uptake of granules increases. Higher water uptake in hydroxypropylated starches is also related to the hydrophilic nature of hydroxypropyl groups . Increasing the oxidant agent and hydrolysis time also led to a significant reduction in the pasting temperature, so that the lowest pasting temperature was obtained in HP30AH24 (49.11°C). In terms of peak viscosity time, native starch had the longest time (5.70 min), and modified starch samples took less time to reach peak viscosity (p < .05). With increasing propylene oxide ratio as well as hydrolysis time, the time to reach peak viscosity decreased (p < .05). The lowest time to reach peak viscosity was observed in HP30AH24 (2.00 min). Native tapioca starch also had the highest peak viscosity  and final viscosity  and was reduced by dual modifying the structure of the starch (p < .05). The effect of hydroxypropylation process on reducing the peak viscosity was greater than the acid hydrolysis so that in hydroxypropylated treatments alone, the peak viscosity was significantly lower than acid hydrolyzed treatments (p < .05). The lowest peak and final viscosities were obtained in HP30AH24 modified starch . Dual modified starches showed lower final viscosity than hydrolyzed and hydroxypropylated starches.The pasting properties of native and modified tapioca starch samples are compared in Table Peak viscosity is defined as the equilibrium point between swelling and solubility (increases viscosity) and the fracture and placement of polymer chains (reduces viscosity). Due to hydroxypropylation of starch, water absorption by granules is facilitated because the hydroxypropyl subgroup is a hydrophilic group and causes more water and starch connections. Increased water uptake and hydrophilic strength after hydroxypropylation increase the amylose release rate mainly in amorphous starch regions, and as a result, the solubility increases . Increasing the oxidizing agent ratio from 10% to 30% caused a significant decrease in To, Tp, and Tc (p < .05). However, acid hydrolysis and its time did not show a significant effect on gelatinization temperatures. The To, Tp, and Tc values of different tapioca starches in this study were in the range of 53.15–63.26°C, 66.61–75.41°C, and 75.25–82.16°C, respectively.The thermal properties of native and modified tapioca starch samples are presented in Table p < .05). In general, acid hydrolysis had no significant effect on the enthalpy of tapioca starches. Enthalpy values of different samples of tapioca starch ranged from 8.75 to 14.40 J/g. The enthalpy of gelatinization is a measure of the quality and quantity of crystals and the loss of molecular order within the granules ; Investigation ; Methodology ; Resources ; Visualization ; Writing‐original draft . Abdorreza Mohammadi Nafchi: Conceptualization ; Data curation ; Funding acquisition ; Project administration ; Supervision ; Writing‐review & editing . Marzieh Bolandi: Project administration ; Supervision ; Validation ; Writing‐review & editing .This study does not involve any human or animal testing.