Ulva species have revealed remarkable variations in genome size due to the integration of exogenous DNA fragments, the proliferation of group I/II introns, and the change of repeat sequences. The genus Ulva is a species-rich taxonomic group, containing a variety of green-tide forming algae. In this study, five complete mitogenomes of the green-tide forming macroalga, Ulva meridionalis R. Horimoto and S. Shimada, were assembled and compared with the available ulvophyceae mtDNAs. The main circular mitogenomes of U. meridionalis ranged from 82.94 to 111.49 kb in size, and its 111.49-kb mitogenome was the largest Ulva mitogenome sequenced so far. The expansion of U. meridionalis mitogenomes is mainly due to the tandem integration of a 5.36-kb mitochondrial circular plasmid (pUme), as well as the proliferation of introns. An intact DNA-directed RNA polymerase gene (rpo) was present in pUme of U. meridionalis and was then detected in two putative plasmids (pUmu1 and pUmu2) found in Ulva mutabilis. The observed integration of the circular plasmid into U. meridionalis mitogenomes seems to occur via homologous recombination, and is a more recent evolutionary event. Many highly homologous sequences of these three putative plasmids can be detected in the other Ulva mtDNAs sequenced thus far, indicating the integration of different mitochondrial plasmid DNA into the mitogenomes is a common phenomenon in the evolution of Ulva mitogenomes. The random incidence of destruction of plasmid-derived rpos and open reading frames (orfs) suggests that their existence is not the original characteristic of Ulva mitogenomes and there is no selective pressure to maintain their integrity. The frequent integration and rapid divergence of plasmid-derived sequences is one of the most important evolutionary forces to shape the diversity of Ulva mitogenomes.Comparative mitogenomics of  Ulva Linnaeus 1753  have attracted much attention not only because of their potential economic value in food and pharmaceutical industry, but also because of their important ecological functions and effects. These green seaweeds widely distributed on the coasts of the world are important indicators to reflect the state of ecological and environmental health. Globally, these opportunistic Ulva species often accumulate large amounts of biomass due to eutrophication, resulting in large-scale green tides    . Organelic level . Based o and II) . In addip to now .Ulva have accumulated rapidly. Thus far, a total of 32 mitochondrial genomes (mitogenomes or mtDNAs) from 19 Ulva species have been sequenced and deposited in the GenBank database  vary greatly in quantity and sequence. It is worth noting that DNA-directed RNA polymerase genes (rpos) are very commonly present in the Ulva mitogenomes, which are likely to be the origin of mitochondrial plasmid DNA, and many rpos have been split into small segments in varying degrees due to multiple mutations  and usually harbor an intronic orf which encoded an LAGLIDADG homing endonuclease (LHE) or a GIY-YIG homing endonuclease (GHE) or a reverse transcriptase/maturase (RTM). Six types of group I/II introns have been found in Ulva mitogenomes, and in particular, the mitochondrial LHEs in group II introns have close relationships with that in group IB introns  have been sequenced and compared with the available ulvophyceae mtDNAs deposited in the GenBank database.To understand the evolution of Ulva meridionalis R.Horimoto and S.Shimada were collected on 4 August 2021 in the Sakura Lake , Rongcheng, Shandong Province, China, when a green tide occurred in the lake due to the proliferation of U. meridionalis (Ume). These Ulva thalli were transported to laboratory in coolers (5–8°C) after collection. Five algal individuals  were randomly selected from 60 identified samples, and named as Ume1 to Ume5, respectively  and a suite of antibiotics  .The drifting algal thalli of ectively . Algal ticillin) . The culrbcL gene)  according to the manufacturer’s instructions. Species identification was performed based on phylogenetic analyses of two common DNA marker datasets  . PrimerscL gene) . SequenccL gene) . The maxcL gene) . The idedionalis , 3.U. meridionalis sample. Clean data were obtained by trimming sequencing adapters and removing short or low-quality reads from the raw data. The complete mitochondrial genomes of U. meridionalis were constructed by the GetOrganelle v1.7.1 (Ulva prolifera (KT428794) was used as a reference genome for assembly. The mitogenome assembly was examined by aligning reads using the MEM algorithm of BWA v0.7.17 , and a Qubit 2.0 Fluorometer , respectively. Qualified DNA samples were fragmented into 350 bp by Covaris S220 ultrasonic crater for library construction. The qualified libraries were sequenced on an Illumina NovaSeq platform  using paired-end sequencing, yielding about 10 Gb sequencing raw data of paired-end reads with 150 bp in length for each e v1.7.1 . The mit v0.7.17 . The Var v0.7.17  and IGV  v0.7.17  were use1 and by aligning the homologous PCGs from the Ulva mtDNAs deposited in the GenBank database with the newly sequenced U. meridionalis mitogenomes. Transfer RNA genes (tRNAs) were searched for by reconstructing their cloverleaf structures using the tRNAscan-SE 2.0 software with default parameters  were found by Open Reading Frame Finder at the NCBI website. Intron insertion-sites were determined manually by aligning the intron-containing homologous genes including atp1, cob, cox1, cox2, nad3, nad5, rnl, and rns. The corresponding genes in the Ulva compressa (KY626327) mitogenome were used as a reference (Protein-coding genes (PCGs) were annotated by Open Reading Frame Finder at the National Center for Biotechnology Information (NCBI) website,rameters . Ribosomeference . Intron eference . The phyeference .rpo) in mitochondrial circular plasmid of U. meridionalis, which we call pUme, was searched against the database of the 98.5 Mbp haploid genome of Ulva mutabilis Föyn  were searched against the NCBI nucleotide database with blastn.The DNA-directed RNA polymerase gene  sequences of full-length settings . The physettings  using MEsettings . InitialU. meridionalis mitogenomes and plasmid DNA was determined using MEGA 7.0  had at least 5000 × depth of coverage  of the Ulva mitogenomes reported , 26 tRNAs, two conserved free-standing orfs, and one putative RNA subunit of RNase P (rnpB)  situated between trnX1 and rpl14 encodes the SecY-independent transporter protein TatC (gene name: tatC or mttB) (trnM3(cau) was detected in the U. meridionalis mitogenomes  which located in the nad4-atp1 and cox3-trnE intergenic regions, respectively. The function of trnX1 is unknown, and the structure of trnS3 is seriously degraded to loss of function in U. meridionalis mtDNAs. On top of that, two copies of specific trnN2(guu) found only in the U. meridionalis mitogenomes are a bit different from the core trnN1(guu) in the sequence of loops, especially DHU loop (trnN1(guu) is very conserved in all known ulvalean mitogenomes. These facts indicate that the trnN2(guu) is likely to appear in the U. meridionalis mitogenome through horizontal transfer rather than trnN1(guu) duplication.The five ogenomes , includiP (rnpB) . One conor mttB) . A conseogenomes , which wDHU loop , while tU. meridionalis, i.e., 14 introns in Ume1 to Ume3 and 13 in Ume4 and Ume5. The intron cox1-731 was present in Ume1 to Ume3 but absent in Ume4 and Ume5 (atp1 (two introns), cox1 (seven or six), cox2 (one), nad3 (one), nad5 (one), and rns (two). Most introns harbored an intact intronic orf encoded an LAGLIDADG homing endonuclease (LHE), or a GIY-YIG homing endonuclease (GHE), or a reverse transcriptase/maturase (RTM) , group IB (GHE), group IIA (RTM), group IIB (RTM), and group II (LHE)  , but in II (LHE) . Five ind before .U. meridionalis mitogenomes reached 20.52–21.65 kb, greatly exceeding the values of all reported Ulva mitogenomes (4.90–19.73 kb). The intronic AT content (61.55–61.97%) was significantly lower than that in mitogenomes (Ulva sp. (MN853878) showed that it contained a total of 22 introns   . These fUme4 and Ume5, as well as the integrated plasmid in Ume1 to Ume3, which we call pUme . The similarity of the AT content between pUme and U. meridionalis mitogenomes might be closely related to their coevolution and plasmid origin, as was markedly different from that observed in plants. Plant mitochondrial plasmids usually exhibit higher AT content than that of mitogenomes, and are suspected to have been acquired from a fungal donor  orf960 and a specific orf424 with unknown function based on blastp search. A small tandem repeat with the period size of 35 bp was located between rpo and orf424. The plasmid-encoded Rpo is in the single-chain RNA polymerase family of phage type that executes a specific-promoter transcription process similar to other multichain RNA polymerases  and rpo/orf961-orf427-rpo/orf945-orf456-rpo/orf161-ψSupres(uua) in WT279 and WT234, respectively  of U. mutabilis, we infer that the reads assembled into these two contigs should belong to mitochondrial plasmid DNA. Two putative circular plasmids named pUmu1 and pUmu2 were constructed, with the size of 4,145 and 11,556 bp, respectively.Based on tblastn search of utabilis , we deteectively . A totalU. meridionalis mitogenomes allowed us to model the integration process of circular plasmid into mtDNAs. The integration of circular plasmid should be via homologous recombination. A 152-bp homologous sequence is shared by orf416 (or orf349) in U. meridionalis mtDNAs and orf424 in standalone circular plasmid DNA, and these sequences are nearly completely identical, except that there is only one base difference at the integration site (the 43rd base of homologous sequences) (orf416 (or orf349) is instead of gtC(V) in orf424. After the first integration of the circular plasmid, two chimeric orfs (orf419 and orf421) were formed. The subsequent integration of circular plasmid should occur at the same recombination site in orf419 which retained the only differential base  . The gtAial base , and genUlva species but not found in mitogenomes of other ulvophyceae lineages  based on blastn search, but they showed varying levels of disruption and degradation in DNA sequence and different types of gene arrangement in structure, indicating the high diversity of plasmids and the differentiation of integration sites via homologous recombination. Sequence homology analysis of these rpos and orfs  showed that they were highly homologous with plasmid-derived rpos and orfs previously found in Ulva mitogenomes (trnY(gua) in all sequenced Ulva mtDNAs, while the homologous suppressor tRNAs of ψSupres(uua) were found in the nad6-trnS1 intergenic region of Ulva rigida mtDNA and in the trnR2-rps3 intergenic region of Ulva flexuosa mtDNAs. These facts showed that the integration of different mitochondrial plasmid DNA into the mitogenome is a common phenomenon in the evolution of Ulva mitogenomes.Many highly homologous sequences of these plasmids  can be detected in mtDNAs of ogenomes . HoweverUlva mitogenomes. In U. meridionalis mitogenomes, almost all specific free-standing orfs originate from plasmid. Although there is no evidence that the orf102 which is located at the cox3-trnX1 intergenic region of U. meridionalis mtDNAs is derived from plasmids, its homologs are present in different intergenic regions of other several Ulva mtDNAs  which happened in rps10 of Ume1 to Ume3 not in that of Ume4 and Ume5. In addition, a 1-bp (A) deletion mutation occurred in the intronic orf575 (RTM) of intron nad3-216 in Ume1, leading to premature termination of its homolog (orf250). A 2-bp (AT) deletion mutation which happened in the latter part of the free-standing orf416 caused the early termination of its homolog (orf349) in Ume4 and Ume5. Intraspecific polymorphisms of five U. meridionalis mitogenomes are significantly high at the intergenic regions and intronic non-coding regions. The divergence in length and sequence of intergenic regions in U. meridionalis mitogenomes was mainly caused by frequent insertion of foreign DNA fragments , rapid accumulation of multiple mutations, and dynamic fluctuation of repeat sequences . A 7-bp sequence (CCTTTGC) is shared by the U. meridionalis mtDNAs and integrated DNA fragment, which should have function in the integration of this fragment.Mitogenomes of is study , but theequences , indicatand Ume5 . The insU. meridionalis mitogenomes, and the order of core genes is almost the same as that of other Ulva mitogenomes (trnE(uuc) from the intergenic region of trnQ-nad5 to that of orf377-trnX1  Rpos, because many of these split Rpos found in lineages . These futabilis , indicatogenomes , indicatogenomes . These morfs from the 39 ulvalean mtDNAs, Ulva species robustly clustered into two primary branches representing two Ulva genetic lineages, I and II, displaying the evolutionary nature of double crown radiation in the phylogeny and speciation of the Ulva group, as is consistent with our previous findings based on phylogenomics analysis of chloroplast genes  first clustered with Ulva sp. (KP720617) and Ulva sp. (MN853878) to form an independent subclade with high support value (100%) , two conserved orfs (orf505 and orf315) and some intergenic regions in this sample are completely consistent with that in U. flexuosa, while other PCGs appear to be chimeras of U. flexuosa (KX455878) and Ulva sp. (MN853878). These results suggest that the mitogenome of this sample is an abnormal chimera, which may represent a cryptic species which we named Ulva sp. (MN861072) in this study.Based on the ML phylogenomic analysis of the aa sequences of 32 genes including 30 PCGs and two conserved st genes . In the e (100%) . An incoequenced , which lflexuosa . The mitUlva species exhibit high diversity at the interspecific and intraspecific level due to their multiple variations which involves the integration of foreign DNA fragments, the acquisition or loss of introns, the dynamic change of repeat sequences, genome rearrangement, and multiple mutations  identified in U. meridionalis and two putative circular plasmids (pUmu1 and pUmu2) detected in U. mutabilis all harbor RNA polymerase genes (rpos) which have only been found in mitochondrial linear plasmids in fungi and plants, which expand our understanding of eukaryotic plasmid diversity. Third, many highly homologous sequences of these plasmids  can be detected in intergenic regions of Ulva mitogenomes sequenced so far, indicating that the integration of different mitochondrial plasmid DNA into the mitogenome is a common phenomenon in the evolution of Ulva mitogenomes. Fourth, these plasmid-derived rpos and orfs have rapidly diverged and degenerated in Ulva mtDNAs, displaying markedly different evolution patterns from the core genes. The frequent integration and rapid divergence of plasmid-derived sequences greatly shape the diversity of Ulva mitogenomes at the intraspecific level. Finally, the ML phylogenomic analysis clearly depicted the evolutionary nature of double crown radiation in the phylogeny and speciation of the Ulva group.The mitochondrial genomes of utations . In thisON402236 - ON402240. Data is publicly available by the following links: https://www.ncbi.nlm.nih.gov/nuccore/ON402236, https://www.ncbi.nlm.nih.gov/nuccore/ON402237, https://www.ncbi.nlm.nih.gov/nuccore/ON402238, https://www.ncbi.nlm.nih.gov/nuccore/ON402239, and https://www.ncbi.nlm.nih.gov/nuccore/ON402240.The data presented in the study are deposited in the GenBank database, accession number FL designed the study, performed the analysis, and wrote the manuscript. FL, HW, and WS performed the experiments. All authors have read and approved the final version of the manuscript.


The start of 2020 provides an opportunity to reflect on 2019 to determine how the South African Journal of Sports Medicine developed. This should be aligned to the journal’s goal to promote research associated with sport and exercise medicine, in particular, research which addresses questions relevant to sport, exercise, nutrition and health in South Africa.In 2019, the journal published eight original research papers, two commentaries, one invited review, one case study, two reports and two conference proceedings. Although the number of outputs is relatively low, on closer inspection, the majority of outputs have direct relevance to the journal’s target audience in South Africa. A journal has to continually find the balance between outputs which are relevant to the target audience and ensuring that these papers are of a high standard. This is a difficult balance to achieve because most authors with high quality work, even with a local relevance, will try and get their work published in high impact international journals. While the South African Journal of Sports Medicine could publish lower quality research papers to keep the number of outputs high, we believe this is not the correct decision. Instead our strategic decision is to maintain the quality of the outputs at the expense of quantity. Less than 50% of the papers submitted for review are accepted. This means that the journal’s growth, defined by the quantity of outputs, will be slower than desired. Growth defined by output quality is more likely to place the South African Journal of Sports Medicine on the trajectory that will result in it getting International Scientific Indexing (ISI) accreditation sooner. This is the desired long-term goal which will enable the journal to become more dominant in this subject field, thereby attracting a greater number of good quality papers. The South African Journal of Sports Medicine is already accredited by South Africa’s Department of Higher Education and Training which enables researchers at South African institutions to get a research subsidy following publication in the journal. The papers published in the journal are also listed by several electronic databases, a full listing of which can be found on our website.The quality of any journal depends on the quality of the reviewers. In most cases, the reviewers have done their job well and provided good quality, thoughtful reviews. This not only assists the authors of the paper to improve their publications, but also raises the standard of the journal. The work by the reviewers is done anonymously and without pay. For this they are thanked in writing by the journal. We have to compete with other journals for reviewers. A mid- to high-tier quality scientist may receive several requests a week to review papers from other journals, so we appreciate the reviewers who agree to assist us. We are planning to have a training workshop during the year to train aspiring reviewers and sharpen the skills of those reviewers who have already some experience.th International Conference on Concussion in Sport will be held in Paris. This is a prestigious conference which defines the current knowledge on sports-related concussion and research for the future. The conference on sports-related concussion started in 2001 and meets every four years. It is supported by the following: International Olympic Committee, Fédération Internationale de Football Association (FIFA), International Federation for Equestrian Sports, World Rugby and the International Ice Hockey Federation. The Sport Concussion Assessment Tool (SCAT) was developed from the conference held in 2005. This tool has become entrenched in sports medicine and is used around the world in a variety of sports. The tool has undergone several revisions to make it a valid and reliable tool and is likely to be tweaked further at this conference. A Scientific Committee coordinates the programme to ensure the knowledge in the area advances. The expert panel at the conference will develop a further Consensus Statement on Concussion in Sport, building on the previous consensus statements which have been published after each conference. ,2 This document will define the management of athletes following a head injury during sport, using the most up-to-date evidence. The progress the scientists participating in the concussion movement have made shows how a focused, well-directed approach can transform concussion knowledge and, in particular, the management of athletes following a head injury. This is a good model for scientists working in other focused areas.The field of exercise and sports medicine continues to evolve. Sometimes the questions with regard to the topics of the subject field are answered and at times the topic loses relevance. Concussion during sport is an area within sports medicine which has attracted scientists from different disciplines. Later this year the 6


In the paper, the method of obtaining the potato starch nanocomposites plasticized with a deep eutectic solvent is described. The deep eutectic solvent based on choline chloride and malic acid  was used as the plasticizer. The effect of the sodium and calcium montmorillonite  addition on the properties of potato starch films was investigated. The thermal, mechanical, and barrier properties were determined. Moreover, a moisture absorption test was performed. The starch gelatinization temperature increased in the presence of montmorillonite. The values of glass transition determined by DMTA depended on the nanofiller type. For the systems containing MMTCa, they generally decreased with its content . The obtained nanocomposites showed improved mechanical and barrier properties. The highest values of tensile strength and Young’s modulus were noted for the system containing 1% MMTNa. The XRD revealed that only the films with MMTNa exhibited intercalation. The homogeneity of the samples decreased with increasing nanofiller concentration. This was probably due to the occurrence of choline chloride-montmorillonite interactions, which were more favored than clay-starch interactions. Plastic pollution is one of the most frequently raised environmental problems. Numerous reports have identified the main sources of pollution. The need to increase the proportion of biodegradable plastics was pointed out . Next toDeep eutectic solvents are eutectic mixtures of two or more components, and their melting temperature is significantly lower (deep) compared to the melting temperatures of the components. This is due to the interactions between the components of the mixture. Currently, five groups of deep eutectic liquids could be distinguished: I: mixtures of quaternary ammonium salts and metal chlorides; II: mixtures of quaternary ammonium salts and metal chloride hydrates; III: mixtures of quaternary ammonium salts and hydrogen bond donors ; IV: mixtures of metal chloride hydrates and hydrogen bond donors; and V: mixtures of non-ionic hydrogen bond acceptors and donors .The drawbacks of thermoplastic starch materials, such as their low resistance to moisture or insufficient mechanical properties, could be overcome by applying nanofillers. According to the shape, a wide range of nanometric structures is known, and includes 1D nanowires and nanotubes, 2D nanosheets, and 3D powders ,8,9,10. In this paper, the method of obtaining the potato starch nanocomposites plasticized with a deep eutectic solvent was described. The deep eutectic solvent based on choline chloride and malic acid  was used. The effect of montmorillonite type , and content on the physicochemical properties  of starch films was determined. Due to the significant differences in suspension viscosity between the montmorillonites, different MMTCa and MMTNa contents were applied. The films based on plasticized starch with MMT were prepared, and the systems containing 1% of the nanofiller were used to compare the effect between the montmorillonites. This kind of material could be considered for applications such as food packaging, edible coatings or in agriculture.gel) corresponded to starch gelatinization and the second one, (Tr) was probably the result of the reaction between starch and the plasticizer . In the case of the starch films containing MMT the signals overlapped, therefore the deconvolution of the thermograms was performed . The higher temperature transition corresponded to the starch glass transition (Tα). In the case of S_CM_MMTCa systems, the Tα generally increased with nanofiller content, but only the sample with the highest content of calcium montmorillonite (S_CM_MMTCa_7) exceeded the Tα of the reference sample. It could suggest a more plasticizer-like activity of the nanofiller. The MMTCa probably formed the aggregates that worked as a lubricant, facilitating the movement of polysaccharide chains (according to one of the theories of plasticization) . B. B−1 assus works ,22,26. TIn In Generally, introducing montmorillonite resulted in a decrease of water vapor transmission rate . For filThe results of the mechanical tests are presented in The sample morphology was analyzed using laser scanning microscopy. The microphotographs of starch nanocomposites are presented in Additionally, the microphotographs of the films with higher montmorillonite content d revealeThe deep eutectic solvent components were choline chloride  and malic acid . Starch was purchased from Zetpezet . Montmorillonites (Ca and Na) were obtained from ZGM .Choline chloride and malic acid (molar ratio 1:1) were placed into a glass vessel and sealed. The mixture was mixed for ca. 0.5 h by magnetic stirrer in a water bath at 80 °C to obtain a homogenous liquid. The eutectic liquid was marked as “CM”.Depending on the montmorillonite type, various nanofiller amounts were introduced into the starch matrix, i.e., for calcium montmorillonite: 1, 3, 5, and 7% (per dry weight of starch), and for sodium montmorillonite 0.5, 1, 1.5, and 2%. The reason was the high viscosity of the MMTNa aqueous suspension. First, the filler aqueous suspension was mixed (500 rpm) for 30 min and subsequently sonified for 10 min . Starch (S) was then added, and the system was stirred for 30 min at 90 °C in a water bath. Next, the deep eutectic solvent (CM)  was added, and the system was stirred for another 10 min (without heating). The starch mixture was cast on a Petri dish and dried at 50 °C. The films were placed in a climate chamber for at least 24 h before testing.The thermal properties of the films were determined by modulated differential scanning calorimetry. The mixture of starch with the plasticizer and nanofiller (ca. 10 mg) was placed in the aluminum hermetic pan. The tests were performed on DSC250 (TA Instruments) in the temperature range from −80 °C to 250 °C, and a heating rate of 10 °C/min (1 cycle).Dynamic mechanical thermal analysis tests were performed using DMA Q800 from TA instruments. The measurements were performed using film tension mode, at a frequency of 1 Hz, a heating rate of 3 °C/min, and a temperature range from −90 to 140 °C. The transition temperatures  were analyzed with TA Universal Analysis software.−2. The spectra were analyzed with OMNIC software.The spectra of starch films were obtained using a Nexus FTIR Spectrometer  equipped with attenuated total reflectance. Each sample was scanned 32 times in a wave number range from 4000 to 400 cmα radiation. The scanning range of the diffraction angle 2θ was from 4.5 to 70°. The d-spacing was calculated from Bragg’s equation, where λ = 0.154 nm.Diffraction patterns were performed on an Empyrean X-ray diffractometer from PANalytical  with Cu K0) and placed in the climate chamber . The samples were weighed (mw) after 1, 3, 6, 24, 72 and 168 h. Moisture absorption was calculated from the following equation:After drying (24 h in 60 °C), starch films (20 × 10 mm) were weighed (m2). The samples were weighed before and 24 h after placing them in a climate chamber . The WVTR was calculated from the following equation:1 is the rate of mass increase in time determined from the graph (mg/h), and S is an area of the tested surface (cm2).The gravimetric dish method was used to measure the water vapor transmission rate (according to ISO 2528 standard). The samples were conditioned in a climate chamber for 24 h before the test. Silica gel was used as a desiccant. The test used an Elcometer 5100 Payne Permeability Cup . At least six replicants of the starch film stripes  were tested. The mechanical parameters  were determined with Bluehill 3 software .The microphotographs of starch samples were performed by a laser scanning microscope model VK 9700  equipped with a violet laser source (wavelength 408 nm) and a pinhole optical system.g values or slightly higher ones than the reference. The XRD measurements were applied to evaluate the degree of MMT dispersion, and only the films with MMTNa exhibited intercalation. The obtained nanocomposites showed improved mechanical and barrier properties. The highest values of tensile strength and Young’s modulus were noted for the system containing 1% MMTNa, i.e., 2.45 MPa and 69 MPa, respectively. The homogeneity of the samples decreased with increasing nanofiller concentration. The microscopic analysis revealed the formation of crystal particles on the surface of the composites containing MMTCa (they were uniformly distributed for lower nanofiller dosage). Interestingly, in the case of the films with MMTNa, the single particles could be noted only for the higher nanofiller content. The probable explanation for the differences in physicochemical properties of the nanocomposites with various nanofillers was the plasticizer-MMT interaction. Such materials can be a potential biofriendly replacement for some of the currently used non-biodegradable plastics, especially for food packaging. Starch films with calcium and sodium montmorillonite were obtained using the casting method. The effect of the montmorillonite type on the starch-based composites was evaluated. The thermal properties of the nanocomposites were tested. The starch gelatinization temperature increased in the presence of MMT. The values of glass transition determined by DMTA depended on the nanofiller type. For the systems containing MMTCa, it generally decreased with its content. The films containing MMTNa up to 1.5% exhibited similar T


The median follow-up was 80 months. Patient & pancreas graft survival, rejection rate, steroid free maintenance rate, HbA1c, weight gain, and the incidence of secondary diabetic complications post-transplant were compared between the groups. p < 0.003 was considered significant (corrected for multiple hypothesis testing). Immunosuppression consisted of alemtuzumab induction and steroid free maintenance with tacrolimus and mycophenolate mofetil. Pancreas graft & patient survival were equivalent in both the groups. BAME recipients had a higher prevalence of type-2 diabetes mellitus pre-transplant , and waited for a similar time to transplantation once waitlisted, although pre-emptive SPK transplantation rate was higher for Caucasian recipients . Despite equivalent rejections & steroid usage, BAME recipients gained more weight , but had similar HbA1c (functioning grafts) at 3-,12-, 36-, and 60-months post-transplant.Ethnic disparities in the outcomes after simultaneous pancreas kidney (SPK) transplantation still exist. The influence of ethnicity on the outcomes of pancreas transplantation in the UK has not been reported and hence we aimed to investigate our cohort. A retrospective analysis of all pancreas transplant recipients (  Despite increasing interest in equitable healthcare, disparities in access to solid organ transplantation, especially for ethnic minority patients, still exists , 2. MostFollowing institutional audit committee approval, a retrospective analysis of all pancreas transplants  performed between January 2006 and March 2020 was done. Data was collected from a prospectively maintained local database and National Health Service Blood and Transplant’s centre database.2. All the DCD donors had a functional warm ischemia time (systolic blood pressure <50 mmHg and/or oxygen saturation of 70%) of less than 60 min and a downtime of less than 30 min.According to our center’s organ acceptance policy, all the DBD donors were less than 65 years old and all the DCD donors were less than 55 years old. The donor’s body mass index (BMI) cut off was ≤30 kg/mImmunosuppression consisted of induction with intravenous alemtuzumab 30 mg (single dose) and methylprednisolone 500 mg. Maintenance immunosuppression was with tacrolimus, mycophenolate mofetil, and a short course of steroids (7 days). Post-transplant target tacrolimus trough levels were 8–12 ng/dl.2, glycaemic lability, and insulin requirement of less than 1 Unit/Kg/day.Patients were classified as type 2 diabetes mellitus if they have a detectable C-peptide and the classification was predominantly based on the diagnosis made by the referring diabetologist. The following is the criteria for transplanting type 2 diabetic patients: Insulin treated diabetes along with end stage renal failure with a body mass index of ≤30 kg/mAll primary and secondary outcomes were compared between Caucasian and BAME recipients; the primary outcome measures were patient and pancreas graft survival, secondary outcome measures were metabolic outcomes among those with a functioning graft , rejection rate, and steroid usage between the two groups.A functioning graft is defined as remaining insulin independent post-transplantation. Secondary diabetic macrovascular complications are defined as any of the following events post-transplant: myocardial infarction, cerebrovascular accident, transient ischemic attack, and/or limb amputations (minor or major). Rejection episodes are either cellular or antibody mediated or mixed, and comprise of either pancreas or kidney rejection (in case of simultaneous pancreas-kidney transplantation). The rejections defined are either biopsy proven or those episodes that were treated based on clinical suspicion .p value of <0.003 was considered significant. All the statistical analyses were performed using Graph Pad Prism software (Version 9.0).Categorical variables are expressed as frequency (%) and continuous variables as median. Differences between the categorical variables were assessed using Fisher’s exact test and differences between the continuous variables were assessed by using Mann Whitney test. Survival analysis was done by using Kaplan-Meir survival plots. Holm-Sidak correction was done for multiple comparisons and a n = 34) were from Asian communities, 30% (n = 16) from Black communities, and 6% (n = 3) from Minority Ethnicities. The median follow-up period of the study was 80 months. The year-wise distribution of pancreas transplant activity in our center is shown in A total of 171 pancreas transplants (SPK-129/PAK-27/PTA-4/Re-transplants-11) were performed during the study period of which 118 recipients were Caucasians and 53 were from the BAME group. Among the BAME group 64%  was significantly higher for Caucasian recipients. Data on workup time, which is the time from referral to our centre to activation in the national transplant waiting list, were available for 45 patients and were similar for both the groups. Data on estimated glomerular filtration rate at the time of referral for SPK transplantation were available for 51 patients and were similar for both the groups. There was a significantly higher prevalence of type 2 diabetes mellitus in the BAME group.n = 6), bleeding (n = 6), sudden cardiac death (n = 3), pancreas failed to perfuse on table (n = 2), severe graft pancreatitis (n = 2), duodenal anastomotic leak (n = 1), Y-graft pseudoaneurysm (n = 1), and unknown (n = 1). The early graft losses were not significantly different between the two groups . The 1-, 3-, and 5-year pancreas graft survival for both SPK and isolated pancreas transplants (PAK/PTA) were comparable between the two groups .The overall rejection rates and steroid-free maintenance rates were comparable between the two groups . The overall incidence of secondary diabetic macrovascular complications post-transplant was not significantly different between the two groups . There were 10 cardiovascular events (Caucasians = 8 vs. BAME = 2), 9 cerebrovascular events (Caucasians = 8 vs. BAME = 1), and 9 peripheral vascular events (Caucasians = 7 vs. BAME = 2).The metabolic outcomes were compared in those recipients with a functioning graft. There were no significant differences in the HbA1c at 3-month, 1-, 3-, and 5-year post-transplant between the two groups . BAME rep = 0.04, and 0.98, respectively). The post-transplant HbA1c among those with a functioning graft were similar for both type-1 and type-2 diabetic patients at 3-month, 1-, 3-, and 5-year . There was no significant difference in the post-transplant median percentage weight gain either .A subgroup analysis was performed to look into whether type 2 diabetes mellitus had an impact on the ethnicity-based outcomes. The overall pancreas graft and patient survival were similar for pre-transplant type-1 and tye-2 diabetic patients  in Caucasian and BAME recipients. This is in contradiction to some of the major studies that have been published before , 4. FromHbA1c among those with a functioning graft was not significantly different between the two groups until 5 years post-transplant, although there is a trend towards higher HbA1c in BAME recipients at 5 years. Longer term follow-up data might uncover the effect of weight gain on HbA1c and graft survival. As HbA1c is known to be an independent predictor of long term pancreas graft failure , other mDespite similar rejection rates and steroid usage in both the groups, BAME recipients gained significantly more weight post-transplantation. In our study we were unable to characterise whether the weight gain was due to an increase in lean body mass or due to increased adiposity. Peripheral hyperinsulinism resulting from systemic venous drainage has been postulated as a cause for excessive weight gain post-SPK transplantation . The expThe incidence of pre-, and post-transplant secondary diabetic macrovascular complications were numerically higher in Caucasian recipients, although, statistically insignificant. There are several reasons for this observation. Firstly, Caucasians had an early onset and a significantly longer duration of diabetes compared to the BAME group. Hence Caucasian recipients had more macrovascular complications due to poor metabolic control. BAME recipients might have had a good metabolic control for a longer period than Caucasians before worsening control and hence the lower incidence of pre-transplant macrovascular complications. Secondly, this is also be due to a more conservative approach in listing Type 2 diabetic patients for pancreas transplantation, as is the case in BAME patients in whom Type 2 diabetes mellitus was more prevalent –31. ExcePrior to 2012, a majority of the patients transplanted were Caucasians and the cold ischemia times were longer. Post 2012, more BAME patients were being transplanted and the cold ischemia time was progressively shorter. Our centre’s change in practice reflected the evolving evidence base as convincing literature evidence was generated around the same time supporting pancreas transplantation in Type 2 diabetes mellitus –34. Due The pre-emptive transplantation rate was higher for Caucasian recipients, although, the estimated glomerular filtration rate (eGFR) at the time of referral, time taken for transplant work-up , and the waiting times were similar for both the groups. There can be several reasons for this. Most of the studies from around the world have reported that lower socio-economic status is independently associated with reduced access to pre-emptive listing –37. DespThis is the first study from the UK reporting ethnicity-based outcomes in pancreas transplantation and the first study reporting metabolic outcomes in Caucasian and BAME patients. Type-2 diabetes was more prevalent in BAME patients. BAME and Caucasian recipients had similar HbA1c until 5 years post-transplantation. Despite similar rejection rates and steroid usage, BAME recipients gained more weight post-transplantation. BAME patients experience similar survival outcomes (graft and patient) to those of Caucasian recipients. Although the waiting time and work-up time were similar, Caucasians had a higher proportion of pre-emptive SPK transplantation.


The present study aimed to evaluate the effect of helium plasma treatment on the wettability of zirconia surface and on the shear bond strength between the dental zirconia core and feldspathic veneering ceramic.  128 zirconia specimens were prepared, polished, and then divided into four groups: control, Zr, FC, and Zr/FC. In Zr and Zr/FC groups, the zirconia blocks were treated by helium plasma for 60 s. In FC and Zr/FC, the feldspathic ceramic powder received 60 s of plasma treatment. Then, the feldspathic powder was applied on the zirconia blocks. Half of the specimens in each group were sintered in a tube furnace, and the contact angle between the zirconia core and feldspathic ceramic was measured at different time intervals. The other half were sintered in a ceramic furnace and then subjected to thermocycling. The shear bond strength was measured using a universal testing machine. The failure mode was assessed using a stereomicroscope. Data were analyzed by one-way ANOVA test, and the statistical significance was considered less than 0.05. P > 0.05). The mean contact angle decreased significantly in all groups over time (P < 0.001). The modes of failure were predominantly mixed in all groups.  There was no significant difference in the mean contact angle and the shear bond strength values of the experimental groups ( The helium plasma applied on either dental zirconia core or feldspathic ceramic powder could not improve the zirconia surface wettability and the shear bond strength between the two ceramics. In recent years, the definitions of esthetic have changed among patients and clinicians . TherefoVarious surface modification techniques have been recommended to improve the bond between the zirconia and veneering ceramic mechanically and chemically such as sandblasting with different particle sizes and different oxides –14, mech4) and argon plasma could significantly improve the zirconia-veneering ceramic bond strength.Recently, plasma technology has been introduced to dental practice, and its potential for improving bond strength has captured researchers' attention; at first, plasma treatment was used to improve the bond strength between the zirconia and resin cement interface –27. LateTo the best of our knowledge, no study has investigated the effect of helium plasma on this bond. Besides, it has been suggested that the helium plasma jet is more efficient compared to argon, since the difference between the metastable state and ionization energy for helium plasma is 3.9 eV which is less than argon ionization energy (4.3 eV) . In otheTherefore, the hypothesis of this study was that the helium plasma might be more efficient in improving the hydrophilic properties of the zirconia surface, thus leading to a stronger bond between the zirconia and veneering ceramic. In addition, due to the higher efficacy of helium plasma jet device compared to argon and the lack of adequate information regarding the effect of helium plasma on the bond strength, the present study aimed to evaluate the effect of nonthermal helium plasma treatment on the wettability of zirconia surface and the shear bond strength between the zirconia core and feldspathic veneering ceramic.The minimum sample size was calculated to be 14 in each experimental group according to a similar study by He et al. , using on = 32) as follows: Control group: zirconia + feldspathic ceramic, both without surface treatment.Zr group: zirconia with plasma surface treatment + feldspathic ceramic without surface treatment . ZirconiFC group: zirconia without surface treatment + feldspathic ceramic with plasma surface treatment. Feldspathic ceramic powder was treated by nonthermal helium plasma for 60 s and the nozzle was in direct contact with the ceramic powder .Zr/FC group: zirconia + feldspathic ceramic, both with plasma surface treatment.From two pre-sintered blocks of zirconia-based ceramic , one hundred and twenty-eight zirconia specimens were cut in dimensions of 10 × 10 × 2 mm by a low speed saw  and then polished with 2500 grit silicon carbide sandpaper . The zirA nonthermal helium plasma device  with flow rate of 10 L/min was used. The frequency and power of the device were 50 kHz and 10 W, respectively .n = 16) to be subjected to the hydrophilicity test and contact angle measurement. 0.05 g of feldspathic ceramic powder  was mixed with the manufacturer's liquid (in a 1 : 1 ratio) and placed on each of the zirconia block using a plastic mold (3.3 mm in diameter and 3 mm in height). Afterward, the specimens were placed in a special tube furnace  at constant temperature of 1500°C up to 120 min with a glass opening which made it possible to take images of specimens during the firing process  with repeated measures test in IBM SPSS Statistics 25 . The statistical significance was considered less than 0.05.P=0.32). Then, the contact angles of samples over time (0 to 120 min) during firing in each experimental group were compared. The mean contact angle values in each group decreased significantly over time (P < 0.001). The contact angle measurements (in degrees) of the experimental groups at different time intervals are summarized in P=0.21). Regarding the shear bond strength values, the highest value was observed in the control group followed by Zr/FC, Zr, and FC groups. However, there was no significant difference in the mean shear bond strength values between the control, Zr, FC, and Zr/FC groups , oxygen (O2), and argon plasma were used and their subsequent effects on the bond strength were investigated.Despite increasing interest in yttria-stabilized zirconia-based all-ceramic restorations in recent years, debonding or chipping of veneering ceramic has been one of the most reported clinical failures associated with full ceramic restorations . As a reIt has been reported that the helium plasma jet is more efficient than argon since the difference between the metastable state and ionization energy for helium plasma is 3.9 eV which is less than that of argon (4.3 eV) . MoreoveFurthermore, the zirconia surface was the only component in zirconia-porcelain restorations that received plasma treatment in the previous studies. It has been suggested that the application of argon plasma reduces the organic matter and generates reactive species on the surface , 32. TheIt is believed that an increase in the surface energy of zirconia might result in more favorable bonding properties . ContactP=0.32). These results were in contrast with those of Ji et al. [The results showed that there was no statistically significant difference in the obtained contact angle values among the control, Zr, FC, and Zr/FC groups during the firing process at different time intervals from 0 to 120 min , as this was expected because of the increased fusion of feldspathic ceramic to the zirconia core over time during the firing procedure in the furnace.Our results also showed that the mean contact angle values in each group decreased significantly over time (Regarding the shear bond strength, helium plasma could not significantly improve the bond strength of veneering ceramic to zirconia core. These results are in contrast with the results of the previous studies , 28, 29 In a study done by Vilas Boas et al. , the sheIn another study, Tabari et al.  evaluateAnother possible speculation is that helium plasma would potentially improve the bond strength if longer exposure time was used. In , the sheIn the oral environment, dental restorations are subjected to thermal and mechanical fatigue and moisture as well. These factors could induce temporary deformation and defects at the material interfaces . ThermocFinally, shear bond strength has some limitations as the predominant mode of failure is usually cohesive within the substrate and not adhesive at the interface. This would indicate that the shear bond strength test cannot represent the actual interfacial bond strength values. However, this was not the case at the present study because the substrate was zirconia, a very high-strength ceramic. Therefore, no cohesive fracture within the zirconia substrate was observed, and the predominant failure was mixed in all the experimental groups such that the fracture was at the interface and then through the feldspathic veneer which partly remained at the interface . These fIt should be noted that the present study is the first study that has evaluated the effect of helium plasma on the contact angle and shear bond strength between the zirconia and feldspathic veneering ceramic. Further studies are required to assess the effect of helium plasma gas using longer plasma exposure times and also the effect of thermocycling on the bond strength between the plasma-treated zirconia and feldspathic veneering ceramic.Within the limitations of the present study, it was concluded that 60 s of nonthermal helium plasma application applied on either zirconia core or feldspathic veneering ceramic powder had no significant effect on the wettability of the zirconia surface by feldspathic veneering ceramic and also on the shear bond strength between the zirconia core and feldspathic veneering ceramic.


The hot compression creep test of the 6082-T6 aluminum alloy sample is carried out by Gleeble-3500 hot compression simulation compressor, and its creep behavior is studied by scanning electron microscope. The results show that the DRX crystal has an irregular shape and that content of the Mg phase, Si phase, and Mn phase in the crystal are the main factors to change the color of DRX crystal. Temperature and strain rate are important factors affecting dynamic recrystallization. Reducing temperature and increasing strain rate will weaken dynamic recrystallization, and DRX critical condition and peak stress (strain) will increase. The constitutive equation of hot creep of 6082 aluminum alloy was established by introducing the work hardening rate-rheological stress curve, and the relationship between DRX critical condition, peak stress (strain) and parameter Z during creep was explored. Based on the Av rami equation, the prediction equation of the DRX volume fraction is established. With the increase of strain, DRX volume fraction is characterized by slow increase, then rapid increase and then slowly increase. In the hot -forming extrusion process of 6082 aluminum alloy, according to the volume fraction prediction equation, the DRX can be reduced, and the internal structure of the material can be optimized by changing the extrusion conditions and particle size. The experimental temperature is 613.15~763.15 K, and the strain rate is 0.01~10 s The results show that dynamic reversion (DRV) and dynamic recrystallization (DRX) are the main mechanisms of microstructure evolution during the thermal deformation of 7A85 aluminum alloy. 350–400 °C is the transition zone from dynamic reversion to dynamic recrystallization. DRV is the main mechanism at 350 °C, while DRX occurs mainly at 400 °C when the sensitivity of microstructural evolution to temperature is relatively high. Li et al. [Al-Mg-Si alloy 6082 has outstanding all-around features, especially high strength, simple processing, and high corrosion resistance ,2. It hai et al.  investigTherefore, it is important to study the dynamic recrystallization behavior of 6082-T6 aluminum alloy and the dynamic evolution of microstructure during thermal creep to optimize the heat deformation parameters and regulate the heat deformation organization and properties. In this paper, a gleeble-3500 thermal simulation compressor, metallurgical microscope (MM) and scanning electron microscope (SEM) are used to investigate the dynamic crystallization and creep behavior of 6082-T6 aluminum alloy based on rheological stress–strain data to represent DRX characteristics. Arrhenius instantonal equation, critical condition model and volume fraction model of DRX are developed. To provide guidance for the thermal processing process of this alloy.−1, respectively, and the specimens were water-cooled immediately after the experiments to retain deformation. After all experiments are completed, the desired specimen is cut along the axial direction. After all the samples were polished, the cross sections of all the samples were analyzed by scanning electron microscope (SEM). The elemental composition of 6882 aluminum alloy is shown in The size of the experimental specimen is Φ8 mm × 12 mm. The specimens were heated to 613.15, 663.15, 713.15, and 763.15 K at 18 K/S by a gleeble-3500 thermal simulation compressor and then maintained for 3 min to make the temperature uniformly distributed, after which the specimens were thermally compressed at strain rates of 0.01, 0.1, 1, and 10 SAs creep proceeds, when the strain reaches a certain value, the stress–strain produces a dynamic equilibrium with a relatively horizontal curve and a distinct dynamic equilibrium ,25. As t−1, 663.15 K, 10 S−1, 713.15 K, 1 S−1, 763.15 K, 1 S−1, as shown in Scanning electron microscope (SEM) was used to carry out microscopic analysis on four samples with test conditions of 663.15 K, 0.001 SComparing The Arrhenius hyperbolic sinusoidal instanton model ,28 was uQ is the material heat deformation activation energy (J/mol); −1); T is the thermodynamic temperature (K); R is the gas constant (8.314 J/mol∙K); and A, a, and n are the material constants.In the above equation, σ for different stress levels.Different power function models are used to describe In the above equation, Taking the natural logarithm of both sides of Equations (1)–(3) to collate gives.The relationship between n = 6.77587 is obtained.From Equation (7), we can find Magnifying Bringing the experimental data into Equation (8), the relation The three terms to the right of the middle sign of the above equation are R (gas constant), the slope of the function To couple the temperature and strain rate on creep behavior, the Zener-Holloman parameter Z  was intrBringing in the relevant parameters obtained in the previous section, we obtain.Taking the logarithm of Equation (10) yields.−1, the intrinsic model of thermal creep of 6082-T6 aluminum alloy is determined.Combining the previously calculated values of The key to DXR and thermal processing optimization technology is to study the critical conditions for DXR. The work hardening rate  yields−1. The corresponding m = 2.75042 and k = 0.101692 at a strain rate of 0.01 S−1 and m = 2.46052 and k = 0.008952 at a strain rate of 10 S−1 are obtained by fitting. Creep recrystallization equations for the corresponding conditions can be obtained by bringing in the relevant parameters.−1 are shown in The predicted creep recrystallization volume fraction curves for 6082-T6 aluminum alloy at strain rates of 0.01 and 10 SWhen the strain rate is constant, the increase in DRX volume fraction is proportional to the increase in temperature; the increase in temperature will give the recrystallization nucleation energy and promote the nucleation to become larger, resulting in the increase of DRX volume fraction. The increase in strain rate will generate a large number of atomic dislocations, resulting in a weakening of the ability to nucleate at the crystallization site, resulting in a decrease in DRX volume fraction. By comparing the metallographic diagrams of the specimens, the recrystallization equations obtained in this paper are in good agreement with the experimental phenomena, indicating that the DRX model of 6082-T6 aluminum alloy can reflect the variation of DRX volume fraction more accurately. Therefore, the creep conditions can be changed to improve the microstructure of the material to achieve the purpose of improving the material properties.The value of thermal creep activation energy Q of 6082-T6 aluminum alloy is 254.473 kJ/mol, and the constructed thermal creep principal equation is:2.−1 are constructed as follows.Based on the Avrami model, the kinetic equations for creep crystallization of 6082-T6 aluminum alloy at 613.15 to 763.15 K with strain rates of 0.01 and 10 S3.Scanning electron microscope analysis shows that DRX crystal blocks have different colors and shapes. When the contents of Mg and Si elements in DRX crystal blocks are far greater than that of Mn elements, the crystal blocks will appear black. When the content of Mn and Si in the DRX crystal block is close, and the content of Mn and Si is far greater than that of MG, the crystal block will appear bright white. When the content of Mn and Si in the DRX crystal block is far greater than that of MG, and the content of Si is greater than that of Mn, the crystal block will appear bright white.4.The critical conditions for creep crystallization of 6082-T6 aluminum alloy were described by introducing the work-hardening rate (θ) and parameter Z. The critical conditions for this material were obtained as follows:


Chlamydomonas under nitrogen (N) starvation. The results showed that the TAG content in FAX1-FAX2-ABCA2 over-expressor (OE) was 2.4-fold greater than in the parental line. Notably, in FAX1-FAX2-ABCA2-OE, the major membrane lipids and the starch and cellular biomass content also significantly increased compared with the control lines. Moreover, the expression levels of genes directly involved in TAG, fatty acid, and starch biosynthesis were upregulated. FAX1-FAX2-ABCA2-OE showed altered photosynthesis activity and increased ROS levels during nitrogen (N) deprivation. Our results indicated that FAX1-FAX2-ABCA2 overexpression not only enhanced cellular lipids but also improved starch and biomass contents under N starvation through modulation of lipid and starch metabolism and changes in photosynthesis activity. The strategy developed here could also be applied to other microalgae to produce FA-derived energy-rich and value-added compounds.Lipid transporters synergistically contribute to oil accumulation under normal conditions in microalgae; however, their effects on lipid metabolism under stress conditions are unknown. Here, we examined the effect of the co-expression of lipid transporters, fatty acid transporters, (FAX1 and FAX2) and ABC transporter (ABCA2) on lipid metabolism and physiological changes in the green microalga  Microalgae are widely regarded as a promising sustainable source for biofuel and bioproduct production ,2. In naIn past decades, microalgae have been regarded as a promising renewable energy source to produce biofuels ,9. HowevChlamydomonas [Chlamydomonas [FAX2 only, the TAG content was not improved [FAX1-FAX2-ABCA2 significantly enhanced TAG accumulation in Chlamydomonas under normal growth conditions [Fatty acids (FA) are the primary building blocks for TAG biosynthesis in plants including microalgae. However, FA biosynthesis and incorporation of FA into TAG occur mainly within the chloroplast and the endoplasmic reticulum (ER), respectively ,14. Duriydomonas ,16,17. Fydomonas . The sinydomonas ,17. Howeimproved ,19. Recenditions . HoweverFAX1, FAX2 and ABCA2) on TAG biosynthesis, lipid remodeling, and cellular responses in Chlamydomonas under N starvation. The results showed that FAX1-FAX2-ABCA2 overexpression enhanced lipid accumulation, improved starch and biomass content, and caused changes in photosynthesis activity. The strategy developed here could be applied to other microalgae to produce FA-derived energy-rich and value-added compounds. In this study, we examined the effect of the co-expression of lipid transporters (Chlamydomonas reinhardtii strain UVM4 [FAX1-FAX2-OE (FAX1-FAX2-35), ABCA2-OE (ABCA2-2), and FAX1-FAX2-ABCA2-OE (FAX1-FAX2-ABCA2-51) were generated in our previous studies [Chlamydomonas cells were cultivated in a tris-acetate phosphate (TAP) medium [−2 s−1) at 25 °C in stackable incubator shakers . To induce nitrogen starvation (48 h), the cells initially grown in the TAP medium were washed and transferred to nitrogen-free TAP media (TAP-N). To analyze the cell number, Chlamydomonas cells were counted using an automated Algae Counter . The ain UVM4  was used studies ,20. The ) medium  with con−ΔΔCT method [RACK1 (Cre06.g278222). The primer sequences used in this study are shown in The total RNA was extracted using RNAiso Plus reagent  Co., Beijing, China) following the manufacturer’s instructions. First-strand cDNA was synthesized using a SuperScript VILO cDNA Synthesis Kit  with 1.0 µg total RNA. Quantitative reverse transcription PCR (RT-qPCR) was performed on Applied Biosystems Quantstudio 5 using PowerUp SYBR green master mix . The 2T method  was usedThe lipids were extracted using the modified Bligh and Dyer method as described previously ,25. To qv/v) and then sonicated three times with a 5-sec interval cycle. The suspension was subsequently autoclaved at 120 °C for 15 min. Enzymic hydrolysis of the starch was initiated by amyloglucosidase (1.5 U) and incubated for 2 h at 55 °C. After the addition of 150 μL anthrone chemical agent, the mixture was subsequently incubated for 10 min at 100 °C, then measured at 621 nm using spectrophotometry . For starch analysis, glucose converted from the starch was quantified using anthrone-sulphuric acid colorimetry as previously described ,28. BrieChlamydomonas cells following the method described previously [The total proteins were extracted from the eviously ,29. The Chlamydomonas cells (20 × 106 cells mL−1) were collected by centrifugation, washed once with 1 × PBS buffer (0.01 M), and resuspended in 500 μL 1 × PBS buffer containing 0.5 μL DCFH-DA. The blank control was made with 500 μL 1 × PBS buffer containing 0.5 μL DCFH-DA. After incubation at 37 °C in the dark for 30 min with gentle shaking, the samples were washed once and resuspended in 500 μL 1 × PBS buffer. The resuspended culture was examined using an automatic microplate reader SpectraMax M2e  for DCF fluorescence detection (excitation at 488 nm and emission 525 nm). The cell-permeable fluorogenic probe 2′,7′-dichlorodihydrofluorescein diacetate  was used to determine the ROS levels as previously described ,30. BrieBriefly, the cell culture (5.0 mL) was filtered using preweighted 47 mm diameter Whatman GF/C filters and dried at 80 °C for 24 h in an incubator. The biomass of the algal cells was then gravimetrically measured as previously described ,30.−2 s−1, 0.8 s). The maximal quantum yield of PS II (v/FmF) and the effective capability of PS II (mΔF/F′) were as we previously described [The chlorophyll fluorescence of the intact cells was measured using pulse-amplitude-modulated fluorometry . Briefly, the cells were dark-adapted for 10 min before applying a saturating pulse  test or the Student’s FAX1 and FAX2 were downregulated by 75% and 83% under N starvation (TAP-N) compared with normal cultivation (TAP), respectively   in lipid metabolism under stress conditions, the expression levels of these genes in the wild-type strain under N starvation were firstly analyzed by quantitative RT-PCR. The results revealed that ectively a. Howevewith TAP a, which  studies ,20, and termined . We founine (PL) b. Strikian in PL b. TAG yian in PL . The larFAX1-FAX2-OE or ABCA2-OE, while the TML increased by 54% in FAX1-FAX2-ABCA2-OE compared with the parental line (PL)  content significantly increased in FAX1-FAX2-ABCA2-OE under N starvation (FAX1-FAX2-ABCA2-OE showed more PUFA and non-PUFA (saturated and monounsaturated) than did any of the control strains. For example, the PUFA content in TFA and TAG increased significantly by 86% and 124% compared with PL, respectively  content in ine (PL) a. TML aline (PL) . The anaE and PL b. For ex with PL b. In FAXABCA2-OE b. These arvation . Noticeaectively c,d. FAX1-FAX2-ABCA2-OE showed different expression patterns between DGAT (diacylglycerol acyltransferases) and PDAT (phospholipid:diacylglycerolacyltransferase) (DGTT1–DGTT4) were significantly upregulated in FAX1-FAX2-ABCA2-OE compared with PL, except that DGTT5 remained unchanged (GPAT (glycerol-3-phosphate acyltransferase) was also upregulated, while that of DGAT1 (type-I DGAT), LACS1 (long-chain acyl-CoA synthetase), and LPAT1 were unaltered or slightly downregulated in FAX1-FAX2-ABCA2-OE compared with PL. Strikingly, in FAX1-FAX2-ABCA2-OE, the genes involved in FA biosynthesis and desaturation, such as ACC1 (acetyl-CoA carboxylase) and KAS1 (3-ketoacyl-CoA-synthase) and FA desaturases , were significantly upregulated  expression was unaltered in FAX1-FAX2-ABCA2-OE  a. Among nchanged a. The exegulated b. These ABCA2-OE a. These FAX1-FAX2-OE and ABCA2-OE, the starch content increased only marginally or remained unaltered compared with the parental line (PL) under N starvation (FAX1-FAX2-ABCA2-OE (FAX1-FAX2-ABCA2-OE showed 1.8-fold more starch than did PL under N starvation (STA1 (the large subunit of AGPase) was considerably elevated in the lipid transporter-OE, while that of STA6  was unaltered compared with PL  and effective photosynthesis capability of PS II (m′ΔF/F) compared with PL, whereas ABCA2-OE exhibited slightly higher v/FmF and m′ΔF/F  level was 7.4-fold higher than in PL  on lipid metabolism during N starvation in the green microalga Chlamydomonas. Moreover, transcriptional and physiological analyses were performed to reveal the potential mechanisms of triple over-expressors of lipid transporters when responding to N starvation. Our results suggest that FAX1-FAX2-ABCA2 overexpression not only has a synergetic effect on enhancing cellular lipid accumulation but also improves starch and biomass content under N starvation. We provide a combined strategy to increase the production of energy-rich carbon reserves in the eukaryotic unicellular microalga Chlamydomonas UVM4 strain. Even though the UVM4 strain is derived from the arginine prototrophic strain cw15-302, it has been used for many microalgal biology and biotechnology studies, such as valuable bioproduct production, protein localization, and recombined protein secretion [Microalgae accumulate abundant triacylglycerols (TAG) under stress conditions, especially during N starvation. Lipid transporters synergistically contribute to TAG accumulation under normal conditions in plants including microalgae ,32; howeecretion ,21. TherArabidopsis thaliana, overexpression of AtFAX1 (FA transporter) or AtABCA9 (ABC transporter) increased TAG accumulation [AtFAX1 were also recently identified and characterized in microalgae, such as Chlamydomonas and Cyanidioschyzon merolae [CrFAX1 and CmFAX1 increased the TAG accumulation in Chlamydomonas and Cyanidioschyzon merolae, respectively [AtABCA9 (CrABCA2) was identified and characterized in microalgae. It was reported that CrABCA2 facilitates TAG accumulation in Chlamydomonas under N starvation [FAX1, FAX2, and ABCA2 are important for oil accumulation, their expression levels were differentially expressed under N starvation  showed more TAG and membrane lipids than did all the control lines under N starvation (DGTT1–DGTT4), ACCase (ACC1), FA synthase complex (KAS1), and FA desaturases  that are involved in TAG and FA synthesis were significantly upregulated in FAX1-FAX2-ABCA2-OE. However, the multifunctional galactolipase PDAT1 was significantly downregulated, and there were no changes in MGDG-specific lipase PGD1  was positively correlated with the increase in starch content [m′ΔF/F and v/FmF are possible reasons for the increased TAG and starch content in FAX1-FAX2-ABCA2-OE under N starvation. Under physiological stress, the lower m′ΔF/F and v/FmF may indicate increasing the availability of reducing equivalents (NADPH) or electrons required for TAG and starch biosynthesis; this will be examined in a future study to decipher the mechanisms involved. Besides TAG and starch, the dry biomass also increased in FAX1-FAX2-ABCA2-OE   a,b. The rivation . In thismponents b, probabmponents c. ROS arondition . Therefod starch . ST68) of Chlorella sorokiniana showed no increase in storage lipids compared to the wild type, which indicates that complicated regulatory steps exist to control carbon re-allocation for lipid and starch biosynthesis in microalgae [FAX1-FAX2-ABCA2-OE showed 1.8-fold more starch than the untransformed parental line (PL), while that in FAX1-FAX2-OE and ABCA2-OE increased marginally or remained unaltered compared with PL under N starvation  showed increased lipid and starch accumulation as well as improved biomass content under N starvation. These results were further supported by the transcriptional analysis of key genes in lipid and starch metabolism.


Additive systematic biopsy (SB) contributes to prostate cancer (PCA) detection in MRI-targeted biopsy (TB). However, the reasons for this are not yet clear. We compared the performance of TB, SB and the combined approach (CB) in biopsy-naive men to determine the added value of SB for tumor grading and spatial tumor distribution.n = 56; 21.6%) were determined. Clinically significant cancer (csPCA) was defined as Gleason grade ≥ 2.Two hundred and fifty-nine men with PI-RADS 3–5 graded lesions who underwent CB were enrolled. Data were prospectively collected, and cancer detection rates (CDR) were compared at patient and lesion level. Gleason grade up- and down-grading from biopsy to prostatectomy specimens (p < 0.001). However, TB showed higher efficiency, defined as CDR per biopsy core, for PCA and csPCA in PI-RADS 4–5 rated patients (p < 0.001). Concordance between biopsy and prostatectomy grading was highest in CB with misdiagnosis of csPCA in 25% of men. TB missed cancer attributed to the index lesion in 10.2% and underestimated csPCA in 7%. In these cases, 76% of csPCA were detected and 85% were upgraded to csPCA by SB in adjacent sectors.CDR by CB based on PI-RADS categories 3, 4 and 5 for PCA were 24%, 72% and 98% and 17%, 64% and 96% for csPCA. CB detected more PCA and csPCA than TB (SB cannot be safely abundant without increased diagnostic uncertainty. When TB missed csPCA, SB detected it close to the MRI-target lesion. Therefore, perifocal biopsies could potentially replace 12-core SB with increased efficiency in taking manageable risks.The online version contains supplementary material available at 10.1007/s00345-022-04230-w. In accordance with prostate cancer (PCA) lethality, discrimination between clinically nonsignificant PCA  and clinically significant PCA  is an important goal. Inter- and intratumoral heterogeneity, however, complicates risk stratification during diagnostic workup. High-quality studies in recent years have impressively demonstrated the potential of the multiparametric magnetic resonance (MRI)-pathway using the Prostate Imaging Reporting and Data System (PI-RADS) in PCA evaluation. Increased detection of csPCA, reduction of nsPCA and beneficially reduced number of biopsies compared to systematic biopsy were shown . NeverthWe conducted this retrospective analysis to clarify the distribution and the diagnostic yields of additive SB for the detection of csPCA during the initial prostate biopsy. Our analysis aimed to investigate the performance of SB, TB and CB and the incremental value of each biopsy method for tumor grading and distribution. Moreover, we investigated the efficiency and the impact of PCA surrogate markers (prostate-specific antigen (PSA), PSA-density (PSAD), prostate volume, abnormal digital-rectal examination (DRE) and abnormal transrectal ultrasound findings (US)) on cancer detection rate (CDR). Finally, we determined spatial tumor distribution according to the MRI index lesion.N = 259 men with PI-RADS lesions ≥ 3 who underwent MRI-targeted and systematic biopsy were included. Uro-radiologists rated and reported the MRI results according to PI-RADS version 2.1. [Biopsy-naïve men who underwent CB were assembled from a prospectively collected institutional database ) and included in the ethically approved (158/22) retrospective audit. Indications for performing MRI were suspicious PSA, abnormal DRE or abnormal transrectal US. ion 2.1. . The majThe same physician performed SB and TB in one session; biopsies were performed by two experienced urologists performing each > 250 prostate biopsies annually . A pre-defined, software-assisted template was used for the institutional standardized biopsy in all patients . Biopsies were conducted under antibiotic prophylaxis, rectal cleansing and local anesthesia. Software-assisted fusion technique (KOELIS Trinity®) was used. All cores were separately documented, collected and histopathologically evaluated. Histopathology was conducted according to guidelines in a reference uropathology [n = 56), we compared the histopathologic results of the prostatectomy specimen with the previous biopsy result.CDR was determined primarily per patient and supplementary per lesion and with respect to the number of cores taken. CDR of SB, TB and CB were compared, stratified by PI-RADS score, prostate volume, DRE, transrectal US findings and PSAD. Lesion-level analysis included the SB results for the MRI index lesion and adjacent sectors. The adjacency was defined as any sector adjacent to the sector with a PI-RADS 3–5 lesion in the transverse or craniocaudal direction. The sectorial division was defined by PI-RADS v2.1 sector map. The subset of patients with divergent SB and TB results in terms of PCA detection, csPCA detection and PCA staging was classified, and tumor distribution was reported. In addition, in a subgroup of our study cohort who had undergone radical prostatectomy at UKB  were reported for continuously coded variables. Differences were detected using the n = 755/3108; 24.3%) and TB . After the procedure, 0.8% of patients suffered a urinary tract infection. More detailed patients’ characteristics are provided in Supplementary Tables 1, 2 and 3.PCA was detected in 71.8% with a prevalence of csPCA in 65.6% and nsPCA in 6.2% of all cases. In total, 993 of 3546 (28.0%) biopsy cores were tumor-bearing, distributed between SB (p < 0.001 for both) and SB , Fig. Highest PI-RADS 3–5-dependent CDR were determined for CB: in detail, 23.8%, 71.5% and 97.5% for PCA and 16.7%, 64.2% and 96.2% for csPCA. Overall CDR for PCA and csPCA by CB were significantly higher than those of TB . Moreover, a predictive value for PCA and csPCA detection by TB and CB was determined for elevated PSAD > 0,1 ng/ml2 and > 0,15 ng/ml2 and decreased prostate volume < 50 ml, all p < 0.001. nsPCA detected by SB was significantly more frequent in men in the absence of suspicious palpation and PSAD < 0.15 ng/ml2 , Supplementary Fig. S2.PCA and csPCA detection by CB, TB and SB were significantly increased in accordance with abnormal DRE and suspicious transrectal US findings (both p = 0.065). The absolute PIRADS 3–5-dependent CDR was 16.7%, 56.2% and 88.7% for PCA and 4.8%, 49.6% and 85.1% for csPCA. Comparing PI-RADS 3–5-dependent efficiency, defined as CDR per biopsy core taken, between biopsy methods, TB showed significantly higher efficiency for PCA and csPCA detection than SB or CB, Fig. TB alone led to cancer diagnosis in 83.3% 155 of 186) of patients and detected less nsPCA (6.8%) than SB (10.8%); however, differences were not statistically significant  of all PCA-diagnosed patients. Despite the absolute increase in cancer detection, no significant differences depending on PI-RADS grades were found for csPCA between SB and TB.TB and SB simultaneously detected PCA in 148/186 (79.5%) patients, but tumor grading by TB and SB matched only in 90/148 (63%) men. PCA, csPCA and nsPCA were solely diagnosed by SB in 32 of 186 (17.2%), 22/170 (12.9%) and 10/16 (62.5%) patients. SB upgraded nsPCA to csPCA in 11/148 (7.4%) or high-risk PCA, defined as ISUP ≥ 4, in 7/148 (4.7%) men. The incremental value of TB for PCA and csPCA detection was 3.2% (6/186) and 2.9% (5/170), respectively. TB diagnosed only one additional man with nsPCA and upgraded nsPCA in 13/148 (8.8%) and high-risk PCA in 5/148 (3.4%) cases.n = 56) matched with previous results of CB, SB, and TB in 68%, 57% and 43%, respectively. TB, SB and CB underestimated tumor grading at 50%, 36% and 30%. Misdiagnosis of csPCA when comparing biopsy and prostatectomy results was significantly lower in CB (25%) than in TB (54%) and SB (60%).Postoperative diagnosis in patients who underwent prostatectomy (p < 0.001 for both). No differences concerning CDR were determined between the predefined locations of SB sampling . In a PI-RADS-dependent analysis, SB cores however revealed a significantly increased CDR for PCA and csPCA comparing men rated PI-RADS 3 (mean 10.8% ± 1.1 and 4.2% ± 2.4), 4 (mean 19.7% ± 3.7 and 11.7% ± 2.8), and 5 (mean 41.7% ± 6.8 and 33.5% ± 6.8), respectively.Significant differences for CDR by TB were shown dependent on index lesion location. PCA and csPCA were more frequently diagnosed in PZ than in TZ , 30/43 (69.8%) csPCA including 4 high-risk PCA were solely detected by SB. In 1/43 (2.3%) suspicious TB negative lesions, SB revealed cancer (ISUP grade 2) within the target area. 23/42 (54.8%) PCA, 22/29 (75.9%) csPCA and 2/4 (50.0%) high-risk PCA were discovered in adjacent sectors of the MRI lesion, respectively. Non-target, non-adjacent cancer was found in 14 cases including 6 csPCA and one high-risk PCA.52/418 TB-detected PCA were upgraded by SB. These 11/52 (21.1%) were upgraded by SB cores on target including three ISUP ≥ 4 tumors. When upgrading was disclosed by SB offside the MRI lesion, 35/41 (85.4%) of the more aggressive PCA were found in adjacent areas and 5/41 (12.2%) were discovered on the contralateral prostate lobe. Hence, TB only approach missed an upgrading to high-risk PCA in 10/41 (24.4%) of the cases, wherein 8/10 (80.0%) were attributed to adjacent sectors and 2/10 (20.0%) to the other prostate lobe, Fig. Before TB can fully replace SB in biopsy-naïve men for reliable detection and exclusion of csPCA, it is important to define the rationale for optimized CDR by SB during the combined approach.In line with previous findings, we showed significantly increased detection rates for PCA and csPCA using CB –10, 13. On the contrary, the underestimation of true tumor grading was high for each solitary biopsy method. Our results corroborate prior prospective series showing the highest concordance with pathological tumor grading for CB (68 vs. 64%) and increasingly diagnostic uncertainties for SB (43 vs. 53%) and TB 57 vs. 47%)  vs. 47% , 16.p < 0.001 for all). These findings imply a high potential to reduce biopsy-related distress, anxiety, pain and infectious risk using the TB-only approach [Advantages of MRI-pathway were also evident in our work: omitting SB could have beneficially reduced the number of nsPCA-diagnosed men by 10/259 and unnecessary core sampling could have been safely avoided by nearly two-thirds. Moreover, TB alone showed a significantly raised efficiency for PCA and csPCA detection in overall and PI-RADS-dependent comparison to SB and CB  csPCA and 50% (2/4) high-risk PCA currently determined by additional 12-core SB. Previously series determined that non-suspicious areas adjacent to the MRI index lesion harbor PCA in 69% and csPCA in 52% of all cases, which is almost similar to our results , 21. The/29 csPCADespite the low efficiency of SB including many unnecessary biopsies, it cannot be eliminated without an increased uncertainty in diagnosis and risk assessment. However, a meta-analysis by Hagens et al. showed a non-inferiority for PCA diagnosis by the TB + regional biopsy approach . Hence, In addition to its retrospective design, limitations of our study include the lack of information on ultimate lesion volume and location on whole mount pathology for the entire cohort. Moreover, no instrumental but a cognitive template was used for standardized institutional 12-core biopsy in all men, which may have led to uncertainty of the sectorial measurement.Nevertheless, our findings demonstrate the promising detection rate of a possible future alternative biopsy method including regional biopsies of MRI-index lesion adjacent PI-RADS map-defined sectors with increased efficiency for biopsy naïve men at a simultaneously reasonable degree of uncertainty regarding csPCA misdiagnosis. The present results hereby emphasize the importance of metric (prostate volume), clinical (DRE/US) and biochemical (PSAD) surrogate markers of PCA in addition to the PI-RADS category to determine the biopsy method of choice on an individual basis.At present, SB cannot be safely abundant without an increase in diagnostic uncertainty. When csPCA is missed by TB, SB detected cancer close to the MRI-target lesion, which suggests that a perifocal saturation biopsy could potentially replace SB with increased efficiency in taking manageable risks.Supplementary file1 (DOCX 413 KB)Below is the link to the electronic supplementary material.


Saccharomyces cerevisiae , were investigated. Although a general response pattern was observed for most of the pasting parameters measured, which includes general enhancements (P < 0.05) in starch gelatinization viscosity, with improvements in gelatinization time and temperature and peak viscosities in most of the varieties, there were significant differences (P < 0.05) in their respective peak time and temperature requirements for the attainment of peak viscosity. Values for the breakdown viscosity were also generally increased (P < 0.05). The technology also increased values for setback viscosity in both the ‘Nkabom’ and ‘Bankyehemaa’ varieties but reduced setback values in the ‘Afisiafi’ and ‘Esambankye’ varieties. As pasting properties are one of the most important characteristics of starch that determine its overall utility, knowledge from this study should inform how adoption of the technology would help diversify the various cassava varieties for appropriate domestic and industrial applications while harnessing its benefits of improved starch yield.Previous work on enzyme application to starch extraction enhanced yield and starch recovery rates as well as modified some physicochemical properties of starches for potential alternative application to industry. The response of the technology, however, showed some sensitivity to variety. The knowledge gap therefore was to establish whether such physicochemical responses (by the technology) to variety affects the pasting parameters of the starches extracted. The pasting parameters of starches extracted from four different cassava varieties , with the aid of crude pectolytic enzymes from the  Cassava is one of the most economically important food crops in the world. Together with yam and potato, it is ranked highest in terms of dry matter production per hectare and is considered the ninth most important source of dietary energy FAO, . Globall2017Manihot esculenta Cranz) varieties ‘Afisiafi’, ‘Esambankye’, ‘Bankyehemaa’ and ‘Nkabom’ all harvested at nine months after planting (MAP) were obtained under a running project at the Department of Agriculture Engineering, K.N.U.S.T. All varieties were planted on the same field of the Agriculture Research Station  and had been subjected to similar edaphic and climatic conditions  were aseptically cultured in the medium for about 8 days. Over the period, protein content of the medium was monitored using the Biuret test and crude pectolytic enzymes were assayed , as in our previous work (Agyepong and Barimah 2017Starches obtained from the optimum treatment combinations (retention time vrs crude protein dosage) for yield were sampled and effects of enzyme treatment on their pasting parameters were investigated. Effects of the crude protein extract (enzyme) dosage vrs retention time on yield (Agyepong and Barimah Paste Stability at 95°C = viscosity at 95 °C – viscosity at 95 °C (after 15 min)Paste Stability at 50°C = viscosity at 50 °C – viscosity at 50 °C (after 15 min)The pasting characteristics of starch samples were determined based on the modification of the Shuey and Tipples  method, Retrogradation (Setback on cooling) = viscosity at 50 °C –viscosity at 95 °C (after 15 min).© 2008. Completely Randomized Design (CRD) was used to organize all treatments. Data were subjected to a one-way ANOVA (unless otherwise stated) and significant differences were tested using the Duncan’s New Multiple Range Test. Unless otherwise stated, all results were presented as mean ± standard values of three replicates (n = 3) and Least Significant Difference (LSD) were also determined for all parameters. Significant differences between treatment means were accepted at P < 0.05 (unless otherwise stated).For all parameters measured, statistical analyses were carried out with SigmaPlot for Windows Version 11.0 by Systat Software Inc.After profiling the crude enzyme extracts for optimal starch yield (Agyepong and Barimah Pectolytic enzyme activity of 4.91 U and specific activity of 4.291units/mg protein (per minute) were recorded in the crude enzyme extract. However, most of the crude extract’s effects on starch pasting properties were due to the presence of endogenous amylases. An amylase activity of 0.293U, with specific activity of 0.257 units/mg, were obtained  treated root mashes of the various cassava varieties are shown in Figs. The effects of enzyme treatment on starch stability during processing are key to determining starch utility. Its effects on the behaviour of other pasting parameters such as its viscosity at gelatinization, time and temperature required for maximum viscosity, pasting temperature and peak viscosity are essential. According to Mayiza-Dixon et al. , gelatinAlthough enzyme treatment did not affect starch viscosity at gelatinization in all varieties Table , the tecPeak viscosities recorded for all starches were generally higher than reported in most literature   Table . Previou5) Table  has sugg5) Table . Althoug5) Table , the var5) Table .P > 0.05) and the ‘Nkabom’ was not affected . Starches from the treated ‘Afisiafi’ and ‘Esambankye’ varieties had lower peak temperatures and time requirements; those from the ‘Bankyehemaa’ were higher  higher values  increased the breakdown value of starches extracted from all cassava varieties  increased (Table P > 0.05) by enzyme treatment (Table P > 0.05) this parameter in starches from the ‘Bankyehemaa’. Given the varied responses, it can be inferred that response of starches to the technology (in terms of starches’ viscosity at 50 °C) was sensitive to variety.Upon cooling to 50 °C (after holding starch samples at 95 °C), there was an increased viscosity in individual starch samples. This was due to the residual amylose undergoing processes that re-associate its moiety to stabilize the paste during the period. Concerning this parameter, starches from the various cassava varieties had different responses: while enzyme treatment reduced the viscosity of starch samples from the ‘Afisiafi’and ‘Esambankye’ varieties Figs.  was signed Table ; ‘Bankyent Table . The valnt Table . Enzyme P < 0.05) in the stability of starches from the treated and untreated mashes were recorded  increased in starches from treated ‘Nkabom’  in starches from the ‘Afisiafi’ and ‘Esambankye’ varieties  by enzyme treatment. For all treated samples, starches from the ‘Afisiafi’, ‘Esambankye’ and ‘Bankyehemaa’ , or starch viscosity at 50 °C, in starches from the various cassava varieties also gave varied responses to enzyme treatment. Cold paste viscosity is a measure of the viscosity of gelatinized starch after holding it at 50 °C. CPV value was significantly  higher values than those from their corresponding treated samples  (Table The setback viscosity of starches from most untreated varieties recorded significantly (y) Table . The smay) Table , would sy) Table  resultedy) Table , eliminay) Table .P < 0.05) the setback values in the ‘Bankyehemaa’ variety  enzyme application, many other processing parameters (required for cooked starch paste) were impacted differently. Enzyme-assisted extraction generally significantly reduced the time for gelatinization in all starches; those from the enzyme-treated ‘Nkabom’ variety were the most positively impacted as it recorded the highest reduction in its time and temperature for gelatinization. Peak viscosity of starches extracted from the treated root mashes of the ‘Afisiafi’ was not affected; however, those from all other varieties were significantly enhanced. The technology greatly improved the peak viscosity of starches from the ‘Bankyehemaa’ variety at a shorter time but required a higher temperature to attain its peak viscosity. The technology also had varied effects on the viscosities of starches (at 95 °C) obtained from the selected varieties but generally reduced the stability of the viscose starch at 95 °C. Starches from the ‘Bankyehemaa’ and ‘Afisiafi’ varieties recorded the highest paste viscosity and stabilities (at 95 °C) respectively. Effects of enzyme application on starch viscosity and stability at cold temperatures (50 °C) also varied suggesting varietal sensitivity to this parameter. This study has therefore shown that application of enzyme-assisted starch extraction has varied effects on the pasting qualities of starches from different cassava varieties. Overall, the variations observed in the pasting properties of the starches can be attributed to the differences in the biochemical and structural architecture of the starch granules from the different cassava varieties. Application of enzyme technology to starch extraction, however, can help modify the starches pasting properties for various uses. Since industries that depend on starch in their business operations have peculiar viscosity profile requirements, it has become important that such industries closely consider these modifications in pasting parameters to inform their choice of variety. The observed modifications in some pasting parameters therefore suggest that the local starch industry could produce starches tailored for different purposes while benefitting from enhancements in starch yield due to the application of enzyme technology.