Cycas revoluta, however the characteristics of Cycas revoluta remain unknown.Starch is major component in the big seeds of Cycad revoluta seeds were systematically investigated, using various techniques.In this study, the physicochemical and structural properties of two starch samples extracted from Cycad revoluta starch showed distinctive characteristics. For physicochemical properties, in the process of gelatinization, the Cycad revoluta starch showed similar viscosity profile to starches of some potato varieties, but Cycad revoluta starch had higher gelatinization temperature. Upon cooling, Cycad revoluta starch formed harder gels than rice starch. For structure, the molecular weight , branching degree and the branch chain length distribution were determined.The amylose contents of the two samples were 34.3 % and 35.5%, respectively. The spherical-truncated shaped starch granules possessed A-type crystallinity, and had an average diameter less than 15 μm. Compared to most commonly consumed cereal and potato starch, Cycad revoluta starch were different in structure from the main-stream starches. Notable differences in some starch traits between the two samples were recorded, which could be attributed to environmental factors. In general, this study provides useful information on the utilization of Cycad revoluta starch in both food and non-food industries.The results suggested that  Among them, Cycas revoluta, the “Sago palm,” is the most cultivated species and mainly planted for uses in ornamental horticulture  was extracted from the seeds of Cycad revoluta grown in Shanghai City, China ; and the other (T2) was extracted from the seeds of Cycad revoluta grown in Guiyang City, Guizhou Province, China .Two starch samples were used in this study. One .2.3.Apparent amylose content was measured according to the method of Li et al. .2.4.Particle size distribution of the starch granules was determined using a laser diffraction particle size analyzer .2.5.The diffraction pattern of starch samples was determined by an X-ray diffractometer . An X-ray tube Cu-Kα (nickel filter) at 40 kV and 40 mA was used as the X-ray source. The diffraction angle (2θ) was 5°-40° with a step interval of 0.02°.2.6.The pasting properties were measured by a rapid visco analyzer , and the data was recorded and processed by the Thermocline for Windows software. Sample preparation and measurement were based on the procedure described previously .2.7.The starch gel formed after RVA test was further analyzed by a TA-XT2i Texture Analyzer  to determine the textural properties, following the previously published procedure .2.8.Thermal properties were measured by differential scanning calorimetry , based on the previously published procedure .2.9.−1 were measured at 4 cm−1 spectral resolution over 32 scans.Fourier transform infrared spectra was obtained via scanning starch samples using a Nicolet iZ-10 FTIR instrument . Starch sample (5 mg) was mixed with potassium bromide (250 mg) and pressed into a film-coated tablet. Wavenumbers from 400 to 4,000 cm2.10.High-performance anion-exchange chromatography (HPAEC) was performed to determine the branch chain-length distribution of amylopectin. Samples was analyzed by a CarboPac PA-100 anion-exchange column  using a pulsed amperometric detector . Data were collected on the ICS5000 , and processed using chromeleon 7.2 CDS .2.11.Gel permeation chromatography-refractive index-multiangle laser light scattering detector (GPC-RI-MALLS) was used to measure the starch molecular weight. The differential refractive index detector  was equipped with two tandem columns . The data was recorded and processed using ASTRA6.1 software .2.12.1H NMR scanning was performed 32 times, at a Larmor frequency of 500.23 MHz. Data were acquired and analyzed using MestReNova. Average DB was calculated with the following equation: DB (%) =  / *100, where I-1,4 is the integrated signal at 5.12 ppm and I-1,6 is the integrated signal at 4.77 ppm, respectively.The Bruker BioSpin GmbH NMR spectrometer equipped with a tempering unit was used to measure the average degree of branching. Starch sample (10 mg) was mixed with 1 mL of deuterated dimethyl sulfoxide-d6 (DMSO-d6) and the solution was thoroughly mixed at 80°C overnight. The mixture was centrifuged at 12,000 rpm. Then, the supernatant was taken and transferred into an NMR tube. The 2.13.p < 0.05).All traits were measured in duplicate. Data analysis was conducted with SPSS 25.0 statistical software program. Significance of differences between the means were determined by the independent t-test analysis  starch (The morphology of ) starch . In cont) starch , 11.3.2.Cycad revoluta starch samples were presented in Cycad revoluta starches were significantly different in D , D , D (0.9), D (0.5) (p<0.05), with mean values of these parameters of T2 were higher than those of T1. D (0.1) of T1 was higher than that of T2, but the difference was insignificant.Particle size distribution of Cycad revoluta starch granules had an average area diameter smaller than 15 μm, and volume diameter less than 20 μm. The T1 and T2 samples had mean area diameter ) of 12.8 μm and 14.5 μm, respectively. D (0.9) were 26.0 μm and 27.3 μm for T1 and T2, respectively. Generally, judging on the value of mean area diameter, the Cycad revoluta starch granules sized similarly to maize, cassava, and sweet potato starch, and smaller than potato starch, but larger than rice starch  with higher AAC also had higher PV, HPV, CPV. According to previous reports, AAC do have significantly positive correlation to HPV and CPV in some cases [, peak (Tp), conclusion (Tc) and gelatinization enthalpy (∆Hg) were 73.7 and 69.8°C, 78.8 and 74.6°C, 85.6 and 81.2°C, and 15.1 and 15.3 J/g, respectively. The two samples varied significantly (p<0.05) in all thermal parameters except ΔHg.Gelatinization temperatures and transition enthalpies of Cycad revoluta starches are within the range of that reported for 163 rice accessions, wherein To, Tp, and Tc ranging from 59.2 to 76.6°C, 66.7 to 81°C, and 71.8 to 87.8°C, respectively (p) varied from 72.5 to 75.7°C .The z-radius of gyration (Rz), number-average molecular weight (Mn), weight-average molecular weight (Mw) and the degree of the dispersion of the molecular weight distribution (Mw/Mn) of Cycad revoluta starches showed similar Mw, but smaller Mn and higher Rz values than rice starches (1 was significantly higher than that of T2 (p<0.05), this is in consistent with the variation pattern in Mw, Mn, Mw/Mn, and Rz between the two samples , branching patterns of the isoamylase-debranched Cycad revoluta starch samples. T2 had more proportions of short-branch chains (53.8% VS 46.5%) but less long-branch chains (27.8% VS 26.7%) and amylose chains (19.5% VS 25.7%) than T1 (p<0.05). Peng et al. (Cycad revoluta starch sample with higher proportion of amylopectin short chains (T2) did have higher PV, BD and lower HD values than that of sample with lower proportion of amylopectin short chains (T1).The ratios of the three peaks were 46.5 and 53.8%, 27.8 and 26.7%, 25.7 and 19.5%, respectively, in the two g et al.  conclude3.11.High-performance anion-exchange chromatography-pulsed amperometric detection (HPAEC-PAD) was used to determine the chain length distribution of amylopectin in this study. The results are shown in Cycad revoluta starches only varied significantly in proportion of A chain.As presented in Cycad revoluta starches were very different from that of wheat and corn, but similar to some rice varieties (Cycad revoluta amylopectin showed much higher proportion of long chains (B2 + B3 chains), but much lower proportion of short chains (A chain) than that of wheat starch . XRD analysis revealed that the Cycad revoluta starches had A-type crystallinity. During gelatinization, the Cycad revoluta starches showed similar viscosity profile with that of some rice and potato varieties, but higher gelatinization temperature than potato starch. The branch chain length distribution analysis revealed that the Cycad revoluta starches were structurally similar to some rice varieties, but totally different from that of corn, wheat and potato starches. In general, the Cycad revoluta starches had distinctive characteristics and its application remained explored in later studies, and the current study provides fundamental information for application of Cycad revoluta starches in food and non-food industries.The structural and physicochemical properties of two The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.KL: conceptualization, resources, supervision, data curation, writing-original draft, and writing-review & editing. TZ: data curation, methodology, and investigation. HR, WZ, and SH: software. YG: methodology. HC: writing-review & editing and funding acquisition. All authors contributed to the article and approved the submitted version.The authors would like to thank Shantou Science and Technology Bureau (grant no. STKJ2021024), Guizhou University Natural Science Project (2020-23), Guizhou University Seed Program (2020-26), and The Opening Foundation of National Laboratory of Hazard Factors and Risk Prevention of Agricultural Product Quality and Safety (2021DG700024-KF202209) for financial support.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Ectopic peritransplant varicosis represents an uncommon cause of late-onset gastrointestinal (GI) bleeding after simultaneous pancreas and kidney transplantation (SPK). We report on a 53-year-old female patient who suffered from recurrent upper GI bleeding seven years after SPK with persistent graft function. Upper endoscopy revealed perianastomotic angiodysplasias, treated by clipping and Argon-Plasma-Coagulation. Repeated endoscopy showed no signs of anastomotic ulcer. With persistent symptoms, computed tomography and angiography revealed extensive ectopic varicosis around the pancreas and duodenal graft. With no signs of portal hypertension, pancreas graft venous outflow impairment or arterio-venous fistula, the origin of variceal formation remained unknown. The extended finding did not allow for endovascular treatment by embolization. Surgery with extensive variceal ligation led to persistent cessation of hemorrhage and maintained stable graft function. In patients with unclear recurrent upper GI bleeding after SPK, one should consider ectopic peritransplant varicosis as an exceptional bleeding cause. If endoscopic treatments fail, angiography should be performed to rule out unusual causes of vascular complications. In case of extensive peritransplant varicosis, surgery may remain the only successful therapy, whenever possible including graft preservation in well-functioning grafts. Simultaneous pancreas and kidney transplantation (SPK) remains the benchmark treatment in selected patients with type-I diabetes mellitus (DM I) and chronic kidney disease (CKD), with excellent long-term patient and graft survival and high rates of insulin independence ,2. SPK iIn contrast, gastrointestinal bleeding (GI) after SPK is less frequent. Late onset of hemorrhage is mostly related to anastomotic ulcers of the graft duodeno-enterostomy, or severe vascular malformations like arterio-enteric fistula or pseudoaneurysms ,5. An evHere we report on a rare case of late recurrent GI bleeding after SPK due to prominent ectopic peritransplant varices, and without the presence of portal hypertension or venous outflow obstruction. We highlight the diagnostic and therapeutic challenges associated with bleeding complications after SPK and present successful graft-preserving surgical management of ectopic peritransplant variceal bleeding eight years after transplantation.Our patient, a 53-year-old woman, underwent SPK in 2014. Intra- and postoperative course was uneventful, with immediate and persistent graft function. Previous history was diabetes mellitus type 1, diagnosed at the age of 27, with unstable glycemia and repeat hypoglycemic episodes, as well as severe diabetic nephropathy present at the time of diagnosis. At the time of transplantation serum creatinine was 493 µmol/l, equal to a glomerular filtration rate (GFR) of 9 ml/min according to the Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equation . Both grSeven years after SPK, the patient developed GI bleeding for the first time. By that time, the patient was on Rivaroxaban (Xarelto®) for hereditary thrombophilia with deep vein thrombosis. Externally performed gastroscopy showed no pathology, however the graft duodeno-jejunostomy was not accessible for examination. Colonoscopy revealed possible signs of previous diverticular bleeding, and capsule endoscopy was inconclusive due to fecal contamination and delayed passage due to diabetic enteropathy. After the initial bleeding stopped and anticoagulation was discontinued, GI bleeding recurred with the persistent need of 1-2 erythrocyte (EC) transfusions every month. Coincidentally, kidney function and general health status slowly deteriorated over time. Recurrent erythropoietin and iron infusions were administered for severe anemia. After one year, a second capsule endoscopy revealed angiodysplasias at the site of graft duodeno-jejunostomy. Thereon push enteroscopy with Argon-Plasma-Coagulation (APC) was performed. Two days after the intervention, the patient suffered from hemorrhagic shock and emergency push enteroscopy revealed severe iatrogenic anastomotic bleeding, which was successfully treated with endoscopic clips and APC.Two months later, during holidays abroad, the patient was admitted to the local emergency department for severe hematemesis with hemorrhagic shock and need for mass EC transfusion. Emergency upper and lower endoscopy were without result, but no push enteroscopy was available there. After repatriation to Switzerland, computed tomography (CT) showed no signs of recurrent bleeding, however, several pathologically enlarged vein branches around the pancreas and duodenal graft were seen Figure .There was no sign of anastomotic portal vein stenosis or thrombosis. Push enteroscopy revealed recurrent neovascularizations at the site of duodeno-jejunostomy, which were treated by clip application and adrenalin injections Figure .There was no sign of graft rejection, with persistent insulin freedom and without elevation of serum amylase levels. That's why we decided against graft biopsy due to the severe risk of bleeding. Decision was made to undergo graft angiography to reveal the cause of the extended variceal formation. Arterial angiography was without pathological finding, excluding an arterio-venous fistula or pseudoaneurysm Figure .Venous angiography revealed numerous veins, also variceal and tortuous veins, around the pancreatic head and duodenal graft draining into the portal vein Figure .One varix was identified to even drain via mesentery to recipients' portal vein. Graft venous outflow was not impaired, with rapid venous drainage via graft portal vein into the inferior vena cava. Due to these extensive findings of peritransplant varicosis we refrained endovascular embolization due to the little expected chance of success. In light of persistent venous thrombosis in the left external iliac vein with the risk of kidney graft involvement, therapeutic anticoagulation with Phenprocoumon (Marcoumar®) was maintained.With ongoing episodes of GI bleeding, unsuccessful endoscopic treatment and little chance for success of endovascular treatment, interdisciplinary panel discussion opted for surgical exploration. 8.5 years after SPK, we performed median laparotomy. Around the pancreatic head and the duodenal graft, numerous varicose veins were found (Figure Pancreatic and duodenal grafts were otherwise normal and the site of enteric anastomosis was without abnormalities. Numerous varicose veins were ligated. Intra- and postoperative course was uneventful and there was no sign of graft venous congestion after these ligations. Since surgery, the patient had complete resolution of symptoms and no more episodes of gastrointestinal bleeding occurred within eight months of follow-up. Graft function was not compromised by surgery as insulin freedom was maintained, and renal function remained stable.This report describes a case of extensive peritransplant varicosis several years after SPK, causing recurrent severe upper GI hemorrhage, in a patient without portal hypertension, venous outflow obstruction or arterio-venous fistula. After failed endoscopic treatments and without the possibility for endovascular therapy, surgery with variceal ligation was finally successful, with sustained cessation of hemorrhage and persistent pancreas graft function. To the best of our knowledge, this is the first case of graft-preserving surgery for recurrent varicose bleeding after SPK. This case highlights the challenges in the management of vascular complications after SPK and illustrates the importance of an interdisciplinary team approach.Late GI bleedings after SPK are overall uncommon and may occur even several years after transplant. These are usually either caused by severe vascular malformations such as arterio-enteric fistulas and pseudoaneurysms, or graft rejection or infection ,5. With Variceal bleeding as a cause of recurrent GI bleeding after SPK is overall rare and to date, only few cases have been reported. Importantly, most reported cases occurred in patients with either portal hypertension  or with However, in our case, the cause of peritransplant varicose vein formation remains unclear. Although portal venous pressure was not measured, there were no indirect signs of liver disease or portal venous hypertension in this patient. Graft portal vein stenosis was excluded by angiography. However, this does not exclude possible damage or compromise of smaller veins within the graft, which is impossible to detect. Other possible triggers of neovascularization, like peritransplant infection, graft pancreatitis, severe rejection, or infectious causes such as cytomegalovirus, were not present in our patient. Formation of varicosis subsequent to extensive endoscopic clipping and APC application is rather unlikely due to the short time interval between endoscopy and CT. However, only one CT scan was performed right before venous angiogram, so there is no possibility for comparison of the variceal formation and size over time. Most likely, variceal formation was triggered by surgical preparation compromising small peripheral venous outflow. Such formation of jejunal varicosis had been previously described in patients after hepatopancreatobiliary surgery ,17, and Overall, treatment of variceal formation after SPK is challenging and to date, no recommendations or guidelines exist. Endovascular therapy plays a pivotal role in the treatment of vascular complications after SPK, above all arterio-venous fistula ,8, graftIn our case, after interdisciplinary discussion, we opted against endovascular variceal embolization. Because of the extent of the finding, with varices both around the graft duodenum and the adjacent jejunum, not only a transpancreatic but also a transhepatic endovascular procedure would have been required for embolization. The significantly increased risk of intestinal congestion or ischemia associated with such an extensive procedure, with only little chance for success in light of numerous and large varices, was ultimately considered unbearable. Importantly, in our patient, angiography could exclude other vascular complications such as arterio-enteric fistula or pseudoaneurysm as causes of bleeding. In our opinion, angiography is of utmost importance in patients with hemorrhage that cannot be treated endoscopically to rule out such complex vascular complications after SPK.In exceptional cases with severe hemorrhage, graft pancreatectomy represents the only therapeutic option ,15. MostIn conclusion, this report describes a rare case of peritransplant variceal bleeding after SPK in a non-cirrhotic patient with preserved graft function in the absence of impaired graft venous outflow, or other complex vascular complications. After failed treatment attempts including recurrent advanced endoscopies and with endovascular therapy considered unfeasible, surgery with variceal ligation was successful. One should consider newly formed ectopic peritransplant varices as a rare cause of late GI bleeding after SPK. Patients with pancreas transplants, where GI bleeding cannot be localized or treated by endoscopy, should undergo angiography to exclude this uncommon etiology.


Correction to: International Journal of Clinical Oncology (2022) 27:1828–1838 10.1007/s10147-022-02232-7In the original publication, under section: Secondary end points, the sentence starting with: “KM estimated…” should read as:KM estimated median DoR was not reached (95% CI 13.9–not reached) in patients who had confirmed responses.In Results section, under heading Secondary end points, the sentence starting with: "Among..." should read as: Among the 31 patients with confirmed response by IRC, median time to response was 1.8 months . In Table The original publication has been corrected.


Cystobacter sp. strain CF23. AmyCf is composed of 417 amino acids with N-terminal 41 amino acids as the signal peptide, and conserved glycoside hydrolase family 13 (GH13) catalytic module and predicted C-terminal domain with β-sheet structure are also identified. Phylogenetic and functional analysis demonstrated that AmyCf is a novel member of GH13_6 subfamily. The special activity of AmyCf toward soluble starch and raw wheat starch is 9249 U/mg and 11 U/mg, respectively. AmyCf has broad substrate specificity toward different types of starches without requiring Ca2+. Under ideal circumstances of 60 °C and pH 7.0, AmyCf hydrolyzes gelatinized starch into maltose and maltotriose and maltotetraose as the main hydrolytic products with more than 80% purity, while maltose and maltotriose are mainly produced from the hydrolysis of raw wheat starch with more than 95% purity. The potential applicability of AmyCf in starch processing is highlighted by its capacity to convert gelatinized starch and raw starch granules into MOSs. This enzymatic conversion technique shows promise for the low-temperature enzymatic conversion of raw starch. Malto-oligosaccharides (MOSs) from starch conversion is advantageous for food and pharmaceutical applications. In this study, an efficient malto-oligosaccharide-forming α-amylase AmyCf was identified from myxobacter  Corallococcus sp. EGB [Kitasatospora [Bacillus halodurans MS-2–5 [Bacillus stearothermophilus [Corallococcus sp. EGB [Malto-oligosaccharides (MOSs) are linear oligosaccharides consisting of around 3–10 α-D-glucopyranosyl units linked by α-1,4 glycosidic linkages. As functional ingredients, MOSs and their derivatives have been widely used in the food, pharmaceutical and other industries, which can be used as sweeteners, preservatives, bulking agents, thickeners, humectants and prebiotics . MOSs camophilus , maltohemophilus  and GH77 sp. EGB . To achi sp. EGB . However8-barrel domain, a small domain protruding out of the barrel as a longer loop between the strand β3 and helix α3 and a C-terminal domain [Thus far, RSDEs originating from bacteria, filamentous fungus, yeasts and plants have demonstrated effective activity toward raw starch . RSDEs hl domain . Even thl domain . Starch-l domain , and dell domain ,15. In tl domain . The domPontibacillus sp. ZY possesses a significantly elevated specific activity of 12, 621 U/mg, enabling the hydrolysis of raw corn starch into G1 (15.4%), G2 (27.2%) and G3 (32.7%) [Corallococcus sp. EGB hydrolyzes raw starch into G2 (41.3%), G3 (30.5%) and G4 (10.2%) [To date, RSDEs have been evaluated in production of ethanol, porous starch, cyclodextrin and MOSs. Among the enzymes under consideration, α-1,4-amylase AmyZ1 from  (32.7%) . Further (10.2%) . These f (10.2%) . Hence, Cystobacter sp. strain CF23, which exhibited sequence novelty compared to the reported amylases. AmyCf showed efficient hydrolysis of soluble starch into G2 and G3 and G4, and also exhibited activity toward raw starch with G2 and G3 as the main products. The objective of this study was to characterize the novel GH13 MOSs-producing amylase from myxobacteria and its potential application in MOSs preparation. Myxobacteria are microbial factories for the discovery of novel metabolites and lytic enzymes, and abundant of enzymes associated with starch, chitin, pectin, cellulose, β-glucan and xylan conversion have been identified, including hydrolases ,20,21,22Cystobacter sp. strain CF23 as the source. The host strains utilized for gene cloning and protein expression were Escherichia coli DH5, Escherichia coli BL21 and Pichia pastoris GS115 . The vector pEFαA was used for protein expression [The cloning of the α-amylase gene was performed using Cystobacter sp. strain CF23 was used as a template for PCR amplification of the predicted α-amylase using the primers AmyCf-F5′-CCGCTCGAGATGGGGCCGCTGGATG-3′ and AmyCf-R 5′-GCTCTAGACTTCGTCCAGACGGCG-3′. The PCR amplifications were performed using PrimeSTAR HS DNA Polymerase , and the obtained PCR fragments were digested with XhoI and XbaI, following by inserting into the same digested sites of pEFαA to obtain the pEFαA-amyCf plasmid. Then, we transformed P. pastoris GS115 with the pEFαA-amyCf-based plasmid and isolated the positive transformants which could grow on YPD plates containing 100 mg/L zeocin . Inducible expression of the amylase AmyCf in P. pastoris was performed as described before [v/v) methanol at 12 h intervals for the whole 96 h fermentation period [P. pastoris GS115 cells was cultured at 30 °C and 200 rpm for 4 days, the cell-free supernatant was collected and assayed for enzyme activity. The protein content was determined using the Bradford [The total genomic DNA extracted from d before . BrieflyBradford . The enzymatic activities of AmyCf toward soluble starch and raw starch were assessed using the dinitrosalicylic acid (DNS) technique by determining the quantity of released reducing sugars. The enzymatic reaction of AmyCf toward soluble starch or raw starch was performed at 60 °C or 50 °C in 50 mM Tris-HCl buffer (pH 7.0), respectively. One unit of α-amylase activity was defined as the amount of enzyme required to release 1 μmol of glucose under the standard assay conditions. w/v, 1.0%), reaction mixtures containing AmyCf and substrates were incubated at temperatures between 30 and 70 °C for 30 min. The thermal stability experiments were performed by incubating the produced AmyCf at various temperatures, and afterwards evaluating the remaining activity under standardized conditions. The determination of the best reaction pH was conducted under normal conditions by employing multiple buffers with varying pH values. Additionally, the pH stabilities of AmyCf were assessed by subjecting the proteins at 4 °C in different buffers for an incubation of 24 h, followed by determining the residual activity. All of the aforementioned processes were conducted in the absence of Ca2+. In order to assess the impact of metal salts on the activity of AmyCf, the enzyme solution was subjected to pre-treatment with 1 mM EDTA at a temperature of 4 °C for 24 h. Subsequently, the sample was dialyzed against a 50 mM Tris-HCl buffer with a pH 7.0 to eliminate any remaining EDTA. The effects of metal ions on the activity of AmyCf were carried out by the addition of various metals with 1 and 5 mM concentration  under standard condition. The residual activity was assessed as described above. Activity without any additive was regarded as 100%. To determine the optimal reaction temperature of AmyCf toward soluble starch or raw wheat starch (w/v) at 60 °C and pH 7.0 . Otherwise, the hydrolytic activity of AmyCf toward raw starches  was also measured at 50 °C in 50 mM Tris-HCl buffer (pH 7.0). The substrate specificity of AmyCf was determined by measuring the hydrolysis activity toward various gelatinized starches , or maltotetraose (G4), maltopentaose (G5) and maltohexaose (G6) in 50 mM Tris-HCl (pH 7.0) at 60 °C for gelatinized starch or 50 °C for raw starch. In order to ascertain the products distribution from the hydrolysis of starches, the supernatant of the reaction mixture was obtained subsequent to centrifugation. Subsequently, the supernatant was subjected to heat treatment at 100 °C for 10 min to inactivate the enzyme. Thin-layer chromatography (TLC) analysis was performed to examine the composition of the products. For TLC analysis, the prepared hydrolysates were spotted onto the silica gel 60 plates  and separated using n-butanol/methanol/H2O  as the solvent system [v/v) with a heating treatment at 90 °C for 10 min. Malt-oligosaccharide with degrees of polymerization (DP) ranging from 1 to 7 were used as the standards. The prepared AmyCf was incubated with different starch samples ), and sequence comparison and phylogenetic analysis of AmyCf with other 241 members of the family GH13 α-amylases were analyzed by MEGA 11. Based on the classification of subfamily, the alignment of the amino acid sequences of AmyCf with the members of assigned GH13 subfamily was performed using ClustalW version 2.0. The signal peptide was predicted using the SignalP 5.0 ), and the isoelectric point (pI) and molecular mass were determined by the Compute pI/Mw tool (https://web.expasy.org/compute_pi/ (accessed on 12 August 2023)). The protein structure of AmyCf was predicted by AlphaFold algorithm [The conserved domains and catalytic residues of AmyCf were analyzed by Conserved Domain Search via the Conserved Domain Database (CDD) of the National Center for Biotechnology Information (NCBI) . A totallgorithm . All data and experiments were presented and performed in triplicate, and mean values were designed with standard deviations. The data were analyzed utilizing Duncan’s multiple range test, which were operated in SPSS software . Cystobacter sp. strain CF23, has been officially recorded in the GenBank database with the accession number OR405308. The nucleotide sequence corresponding to the amylase gene amyCf, which was obtained from Cystobacter sp. strain CF23 and accordingly named AmyCf, which is composed of 417 amino acid residues. Sequence analysis showed that AmyCf contains a N-terminal putative signal peptide, GH13 catalytic module and C-terminal β-sheet domain, which shared the general structural feature with family GH13 α-amylase [Cystobacter fuscus. Subsequently, the α-amylases originating from Archangium and Stigmatella hybrida displayed a sequence identity of 84%. However, these proteins from myxobacteria are annotated as predicted α-amylase family without biochemical characterization. By genome comparison analysis, a potential Malto-oligosaccharides-forming amylase was identified from Corallococcus sp. EGB [Massilia timonae CTI-57 [Currently, α-amylase enzyme specificity within the CAZy database can be found unambiguously in the families GH13, GH57 and GH119; the family GH13 is considered to be the main α-amylase family first proposed in the CAZy system . In gene sp. EGB  and the e CTI-57 , which se CTI-57 , and thee CTI-57 ,31, indiCorallococcus and Archangium. AmyCf from Cystobacter shares low sequence identity (<30%) with these enzymes except for AmyM (42%), and AmyCf is the first identified α-amylase from Cystobacter. Phylogenetic analysis showed that AmyCf and amylase AmyM from myxobacteria form an independent branch, which is separated from the known MOS-producing amylases from other microbes  have been employed for the purpose of maltose conversion. However, it is worth noting that their ability to hydrolyze raw starch is constrained. Otherwise, most of the reported amylases operate in an endo-lytic manner, a characteristic that influences the presence of glucose in the resulting products. However, feasibility of MOSs is dependent on their DP values . The preCystobacter sp. strain CF23 with hydrolytic activity of raw starch, which shares low sequence identity (<45%) with the reported amylases. Under the optimal conditions, the special activities of AmyCf toward soluble starch and raw wheat starch are identified as 9249 U/mg and 11 U/mg, respectively. AmyCf effectively hydrolyzes gelatinized starch into G2, G3 and G4 without depending on Ca2+. It is noteworthy that AmyCf also displays the ability to hydrolyze raw starch into G2 and G3 with the absence of glucose. The uniform hydrolysis and diverse breakdown abilities makes AmyCf a highly viable choice for the enzymatic hydrolysis of starch into MOSs. Further research is warranted to explore the catalytic mechanism of AmyCf involved during hydrolysis processes, with particular emphasis on understanding the substrate selectivity. In this study, we identified a special Malto-oligosaccharides-producing amylase AmyCf from


Cystinosis, a rare lysosomal storage disease caused by mutations in the CTNS gene, is characterized by cystine crystallization and accumulation within multiple tissues, including kidney and brain. Its impact on neural function appears mild relative to its effects on other organs during early disease, but since therapeutic advances have led to substantially increased life expectancy, neurological implications are of increasing interest, necessitating deeper understanding of the impact of cystinosis on neurocognitive function. Behavioral difficulties have been reported in cystinosis in the visual domain. Very little is known, however, about how the brains of people living with cystinosis process visual information. This is especially interesting given that cystine accumulation in the cornea and posterior ocular structures is a hallmark of cystinosis.Here, high-density scalp electrophysiology was recorded to visual stimuli (during a Go/No-Go task) to investigate visual processing in individuals with cystinosis, compared to age-matched controls. Analyses focused on early stages of cortical visual processing.The groups differed in their initial cortical response, with individuals with cystinosis exhibiting a significantly larger visual evoked potential (VEP) in the 130–150 ms time window. The groups also differed in the associations between neural responses and verbal abilities: While controls with higher IQ scores presented larger neural responses, that relationship was not observed in cystinosis.The enlarged VEP in cystinosis could be the result of cortical hyperexcitability and/or differences in attentional engagement and explain, at least partially, the visual and visual-spatial difficulties described in this population.The online version contains supplementary material available at 10.1186/s13023-023-02985-y. Cystinosis is an autosomal recessive disorder caused by bi-allelic mutations in the 17p13.2-located CTNS gene , 5, charTo reduce this gap in knowledge, our research team has focused on characterizing the impact of cystinosis on brain and cognitive function. Though abnormally high levels of cystine have been observed in different brain regions –14, cystHere, we expand this investigation to the visual sensory domain. Interestingly, visual-perceptual indices are often significantly lower than verbal indices in individuals with cystinosis –19. ThisHere, we use high-density EEG to investigate basic visual processing in a group of individuals with cystinosis and compare them to a group of age-matched controls. The analyses focus on the visual evoked potential (VEP) component P1. P1 is an early VEP peaking around 100 ms following stimulus onset and has been associated with multiple generators in both dorsal and ventral visual streams –33. AddiConsidering the behavioral difficulties reported in visual-related processing in cystinosis and the tendency for cystine accumulation in the retina, we hypothesized that individuals with cystinosis would show different sensory-perceptual brain responses to visually presented stimuli, reflected in amplitude differences in the P1 component of the VEP. Identifying the processing stages that are impaired and contribute to visual and visual-spatial processing differences in cystinosis will be important in the development of impactful strategies that address visual processing difficulties and identify sensitive biomarkers of treatment efficacy on brain function.Thirty-eight individuals diagnosed with cystinosis  and 45 age-matched controls  were recruited. Individuals with cystinosis were recruited via social media and through contact with family organizations. Due to the rareness of cystinosis, most participants, all of whom lived within the United States, traveled from out-of-state to participate. Furthermore, the controls were recruited via flyers in the neighborhoods surrounding the lab and through a lab-maintained participant database. Developmental and/or educational difficulties or delays, neurological problems, and a severe mental illness diagnosis were exclusionary criteria for controls. Current neurological problems were exclusionary criteria for individuals living with cystinosis. To assess visual acuity, a Snellen chart was used. All participants had normal or corrected to normal vision. Nevertheless, all participants were asked at the start of the electrophysiological experimental session if they could see the stimuli and their different components without difficulty. One individual with cystinosis was excluded from the final sample due to illness on the scheduled day of testing. All individuals, and their legal guardian if under 18 years old, signed a consent form. Participants were monetarily compensated for their time. This study and all the associated procedures were approved by the Albert Einstein College of Medicine Institutional Review Board (IRB 2009-523). All aspects of the research conformed to the tenets of the Declaration of Helsinki.Participation consisted of two visits, which involved completion of a cognitive function battery and EEG recordings. The cognitive function battery included the assessment of verbal and non-verbal intelligence  and executive functioning components  were presented in a pseudorandom sequence. Participants were instructed to press the left mouse button with the right index finger upon each stimulus presentation, as quickly and as accurately as possible, unless the stimulus was a repetition of the immediately preceding stimulus, in which case they should withhold their response . Stimuli, subtended 8.6° horizontally by 6.5° vertically, were presented centrally for 600 ms at an average rate of 1 per second . Three 12-min blocks were run. Each block consisted of 540 trials, for a total of 1620 per participant. Here, we focus exclusively on hit trials, that is, trials that included a correct button press after stimulus presentation (only hits preceded by another hit were included), to ensure a good signal-to-noise ratio and to avoid overlap of processes related to inhibiting a response .https://github.com/DouweHorsthuis) [Continuous EEG data were recorded from 64 scalp electrodes at a sampling rate of 512 Hz  and preprocessed using the EEGLAB toolbox (version 2021.0)  for MATLrsthuis) . Preprocp-values (from t-tests and Spearman correlations) were submitted to Holm-Bonferroni corrections for multiple comparisons [p.adjust of the stats package in R [lmer function in the lme4 package [p-values were estimated using Satterthwaite approximations.P1 was measured between 130 and 150 ms at O1, Oz, and O2. Time windows and electrode locations were selected based on past research and confirmed (and adjusted) by inspecting the timing and topography of the major voltage fluctuations in the grand averages. Mean amplitude data were used for both between-groups statistics and Spearman correlations. All parisons , using tage in R . Mixed-e package  in R [40Welch corrections were applied to adjust the degrees of freedom. A chi-square test was run to test for independence between sex and group. Effect sizes were calculated utilizing Cohen’s d and w. As can be seen in Table Table ß = 7.55, SE = 2.45, p = 0.01).The averaged VEPs for P1 per channel and by group can be seen in Fig. rs = −0.73, p = 0.01). As can be seen in Fig. rs = 0.40, p = 0.04), such correlation was not significant in cystinosis. P1 did not correlate significantly with non-verbal IQ, for either the control or the cystinosis group.Figure Despite ample evidence for atypical visual processing in cystinosis in the neuropsychological literature , 26, 42 Increased sensory-evoked potentials may reflect increased excitability in sensory cortices –47. An iAlthough we think it unlikely, one cannot rule out the possibility that group differences in P1 amplitude, alternatively, reflect different levels of attentional engagement. Selective attention, most often in the context of visuo-spatial attention designs, has been shown to significantly modulate P1 amplitude –56. SuchP1 has been localized to sources in the dorsal extrastriate cortex, specifically to areas V3, V3a, and adjacent middle occipital gyrus and in the ventral extrastriate cortex, specifically area V4 in the fusiform gyrus –32 and iThe increased amplitude VEP in cystinosis appears to be especially localized to the occipital and lateral parietal-occipital channels. While the current analyses focused on the channels in which response was maximal for both groups, those channels also represented the largest differences between the groups. As one can appreciate in Additional file Correlational analyses revealed that neural indices of visual sensory processing correlated with age in a similar fashion across groups. In the general population, VEPs attenuate in amplitude through at least late adolescence . Here, P1 attenuation was replicated in our control sample and was also observed in the cystinosis group. The groups differed, however, in the correlations between P1 and verbal IQ. While controls with higher IQ scores showed larger P1s, a significant relationship between P1 and verbal IQ was not present in the cystinosis group. P1 did not correlate significantly with non-verbal IQ, for either the control or the cystinosis group.Lastly, a brief detailing of findings relating to general cognitive function in cystinosis is merited. Neurocognitive assessments showed lower verbal and non-verbal IQs in individuals with cystinosis, when compared to their control peers. We and others had previously reported lower IQ scores in this population , 19, 23 This study is not without limitations. First, our groups were not matched in terms of IQ, with the cystinosis group presenting significantly lower scores than the control group. Although there is no evidence of the association between VEPs and IQ, such differences could have impacted the results. Second, variables related to current health status  and compliance with treatment, which has been linked to better clinical outcomes , were noTo conclude, here we present evidence of visual processing differences in cystinosis which could contribute, at least partially, to both photosensitivity and to the behavioral visual-spatial difficulties described in this population. More work is needed to describe how visual processing differences might contribute to the cognitive profile associated with cystinosis. A better understanding of such associations could contribute to the identification of sensitive biomarkers of treatment efficacy on brain function. The current findings should motivate future studies utilizing paradigms tapping into higher- and lower-order visual processes and investigating visual processing and pathways in more depth, for instance, by employing tasks distinguishing between ventral and dorsal streams and magnocellular and parvocellular pathways. However, in so doing we believe it is important for future EEG research with this population to take sensitivity to light into consideration when designing visual experiments. Furthermore, these findings and their implications in classroom and professional settings utilizing digital media and screens should be further explored.Additional file 1. Averaged ERPs per group over parietal, parietal-occipital, and occipital channels.


The omental flap is often used to fill the space around the artificial vascular graft as a network sheet to prevent artificial vascular infection. In this study, we report a case in which the omental flap was divided into three parts to fill the dead spaces around the multiple-branched graft, as well as to wrap the suture lines of the graft after graft replacement in a patient with an infected thoracic aorta.An 88-year-old woman was admitted to the hospital with fever and impaired consciousness. Computer tomography revealed an aortic arch aneurysm with enlargement. After emergency stent–graft interpolation and antibiotic treatment, an infected thoracic aortic aneurysm was removed, and a multiple-branched graft replacement of the upper arch was performed. After harvesting an omental flap based on the right gastroepiploic vessels, the omental flap was divided into three on the basis of the epiploic vessels. The middle part of the omental flap was used to fill the space around the lesser curvature of the arch and the distal anastomotic site, the accessory part was used to fill the space between the ascending aorta and the superior caval vein, and the right part was used to wrap the three cervical branches, separately. Fifteen months after surgery, the patient had recovered enough to resume work without any signs of inflammation. The aneurysm is often replaced by an artificial vascular graft, which is covered by an omental or muscle flap to prevent graft infection wherever possible.Escherichia coli was cultured from the urine and blood, and ceftriaxone was administered. Emergency stent–graft interpolation was performed 1 week after hospitalization, and the infection subsided after treatment with piperacillin/tazobactam.An 88-year-old woman was admitted to the hospital with fever and impaired consciousness. Computer tomography revealed an aortic arch aneurysm with enlargement. Five weeks after admission, the infected thoracic aortic aneurysm was removed, and a multiple-branched graft replacement of the upper arch was performed A. We harThe omental flap is often used in patients with mediastinitis or infected aortic aneurysm. Moreover, it is often used to fill dead spaces as a network sheet without division. Settembre et al.Kim et al.The omental flap can be maximally utilized by confirming flap blood flow and dividing the flap into multiple parts as needed.None.Not required.None.


The first uses polyurethane (PU) as the polymeric binder and the second uses polydopamine (PDA). Our antimicrobial PU/Cu2O and PDA/Cu2O wraps, respectively, kill >99.98% and >99.82% of the human pathogen, P. aeruginosa, in only 10 min, and each of them kill >99.99% of the bacterium in 20 min. These antimicrobial wraps can be removed and replaced on the same object in <1 min with no tools. Wraps are already frequently used by consumers to coat drawers or cars for aesthetic or protective purposes.Antimicrobial coatings have a finite lifetime because of wear, depletion of the active ingredient, or surface contamination that produces a barrier between the pathogen and the active ingredient. The limited lifetime means that facile replacement is important. Here, we describe a generic method for rapidly applying and reapplying antimicrobial coatings to common-touch surfaces. The method is to deposit an antimicrobial coating on a generic adhesive film (wrap), and then to attach that modified wrap to the common-touch surface. In this scenario, the adhesion of the wrap and antimicrobial efficacy are separated and can be optimized independently. We demonstrate the fabrication of two antimicrobial wraps, both using cuprous oxide (Cu The human and economic burdens of infectious disease are vast ,2,3,4,5.The most common strategy to inhibit disease transmission via fomites is to disinfect solid surfaces by wiping them with bleach or 70% ethanol, but it is time-consuming to wipe many surfaces, and some disinfectants  are hazardous . PerhapsAntimicrobial coatings can be either incorporated during the manufacture of an object or applied to the finished object. Application to existing facilities is particularly important for a rapid response to evolving situations such as the COVID-19 pandemic: it is expensive and time-consuming to replace many common-touch items  in a short period of time. Ideally, the installation of antimicrobial coatings in existing facilities should be rapid and should not require sophisticated technical skills or equipment so that it can be carried out by non-experts on site.In this manuscript, we describe a method for rapidly applying an antimicrobial coating to common-use objects. The method is simple. We prepare an antimicrobial coating with an adhesive backing layer that can be simply applied to a solid by removal of the backing paper, application to the solid, and sometimes followed by moderate pressure onto the solid. This type of coating is common for cars, where it is called a wrap, or for lining drawers, where it is called contact paper, and is available in sheets with dimensions of meters. The primary criteria for the preparation of the antimicrobial wrap are that application of the antimicrobial layer does not degrade the integrity of the wrap and does not reduce its ability to adhere to surfaces. The use of a wrap also solves an important problem with antimicrobial coatings. Antimicrobial coatings have a finite lifetime because of wear, depletion of the active ingredient, or surface contamination  that produces a barrier between the pathogen and the active ingredient. When the coating is no longer functional, the coating must be cleaned (to remove contamination) or removed and replaced with a functional coating. The removal step can be time-consuming as is well-known for repainting houses, etc. For situations where the coating must be replaced frequently, the time for replacement is a significant cost, whether or not the old coating must be removed. In situations such as these, a wrap is very efficient. When the old coating is compromised, the wrap is peeled off, and a new wrap is applied. The adhesive properties of the wrap can be optimized for easy removal or longevity, depending on the application. In general, the adhesion and the antimicrobial layer can be optimized independently.2O) [2O has broad antimicrobial activity against many pathogens, for example, against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, and Enterococcus hirae [S. aureus (MRSA) [2O using previously developed coatings [This work describes two antimicrobial wraps that allow the very easy conversion of a common-touch surface into an antimicrobial common-touch surface. Both wraps utilize a potent antimicrobial material: cuprous oxide (Cu2O) ,31. Cu2Ous hirae ,32,33, as (MRSA) ,19. The coatings ,19,20 th2O wraps against P. aeruginosa. P. aeruginosa is labeled by the United States (US) Center for Disease Control (CDC) as a serious threat [P. aeruginosa has developed antibiotic resistance to many antimicrobials [We provide measurements of the bactericidal activity of the Cus threat  because crobials ,36. Of pcrobials  and formcrobials .2O wrap), which caused a 99.97% reduction in P. aeruginosa colony-forming units (CFUs) in 10 min and >99.99% in 20 min. The second used polydopamine (PDA/Cu2O wrap) which caused a reduction of 99.80% in 10 min and >99.99% in 20 min. Polydopamine (PDA) has numerous advantages: it is derived from natural sources, uses water as the polymerization medium, has a high LD50 (~500 mg/kg), and polymerizes on most solid materials [To illustrate the versatility, we prepared two antimicrobial coatings on the same commercial wrap, which was adhesive-backed polyethylene (PE). The first wrap was prepared using polyurethane , and then, a suspension of Cu2O in ethanol was cast on the tacky PU layer. The sample was heated for 2 h to accelerate the cure of PU. Scanning electron microscopy (SEM) images of the coating are presented in 2O coating only had a thickness of 10–20 µm, and the loading of particles in the PU/Cu2O wrap was 4.5 × 10−3 g/cm2. Because the coating is thin, only a very small amount of copper is used per area, which reduces the cost of implementation.The fabrication of both wraps is uncomplicated. The PU/Cu2O wrap was prepared by applying a suspension of Cu2O in dopamine to the uncoated wrap, and then, it was heat-treated to complete the polymer cure. 2O wrap. As shown in 2O particle morphology is visible, indicating that the particles protrude from the coating and therefore are accessible to solutions and bacterial suspensions. The particles form a much sparser layer on the solid compared to the PU/Cu2O wrap. The PDA/Cu2O wrap has a loading of 0.13 × 10−3 g/cm2 of Cu2O particles, which is about 1/35th of the loading of PU/Cu2O wrap, thereby enabling even lower costs of the active ingredient.The PDA/Cu7 CFU/mL P. aeruginosa on each wrap, waiting for a preset interval , and extracting the bacteria from the solid, followed by counting the colonies formed by bacteria that remained viable (CFUs). The resulting CFUs were compared to the input CFUs to obtain the log survival, as defined by the following equation:The antimicrobial activities of the wraps were tested by placing a 5 µL droplet of approximately 1.6 × 10Both wraps caused a large decrease in the bacterial viability in a short period of time.2O wrap, as shown in P. aeruginosa in 10 min, and >99.99% (4 log reduction) in 20 min. For registration, the EPA requires an antimicrobial coating to cause a minimum 3-log (99.9%) reduction in 1 h; the wrap achieved a greater reduction in only 10 min. Using the 99.9% reduction from the control as our metric, we performed Student’s t-tests separately on the 10 min and 20 min data, comparing 1/1000th of the control CFUs to the wrap CFUs to show that the antimicrobial wrap caused a reduction of >99.9% (p < 0.001) compared to the uncoated wrap after 20 min, but that this target was not achieved in 10 min (p = 0.22).For the PU/Cu2O coating described above included heat and plasma treatments, whereas for simplicity, it would be beneficial to prepare a coating that does not require these steps. We prepared a second set of PU/Cu2O coatings with no heat or plasma treatment. As shown in p = 0.003) for 20 min of exposure.The preparation of the PU/Cu2O wrap was also extremely effective against P. aeruginosa, as shown in 2O wrap. This shows that the Cu2O is a very potent antimicrobial, and it is unnecessary to provide the high loading of the PU/Cu2O wrap to achieve a good performance in these tests.The PDA/Cu2O and PDA/Cu2O wraps were designed for rapid replacement. 2O wrap was added and then replaced. The replacement of the coating took less than 1 min and was achieved without tools.Recall that the motivation for this work is the simple replacement of an antimicrobial coating to overcome the problems of wear and contamination. Both the PU/Cu2O coating used ethanol, which is a relatively safe solvent. The polymerization of dopamine only requires water as the solvent. Both coatings require a low temperature to complete the cure (80 or 120 °C), and we found that heating was not even required for the PU coating. In any case, this heat treatment is not required on site but is only used for manufacturing. Paintable PU formulations have some toxicity due to the evaporation of solvents, but this need only be carried out during the manufacture of the wrap, not at the implementation site. The Median Lethal Dose (LD50) of polydopamine is high, about 500 mg/kg [2O wrap (no plasma or heat-treatment) that was tested against P. aeruginosa remained active against the bacterium. Finally, the processes used to prepare both antimicrobial wraps do not require complex equipment.Here, we discuss characteristics of the specific coatings that we used on the wraps. The PU/Cu00 mg/kg . CuprousThe utilization of antimicrobial coatings has been shown to correlate with a lower number of infections ,29. Howe2O. One utilized polyurethane and the other polydopamine as the binding polymer. Both coatings killed >99.995% of P. aeruginosa in 20 min, and the bacteria had half-lives of <1.4 min. By demonstrating the preparation of two antimicrobial coatings on the same base wrap, we showed that the wrap concept is modular, which allows the independent optimization of the adhesive coating and the antimicrobial surface.For the highest effectiveness, an antimicrobial wrap needs to kill microbes rapidly. We prepared two different antimicrobial wraps based on the active ingredient Cu2O particles  were obtained from American Chemet Corporation . Polyurethane  was obtained from Lowes Home Improvement departmental store . TSA (Tryptic Soy Agar) and TSB (Tryptic soy broth) were purchased from BD, Sparks, MD, USA.In this study, 70% ethanol was purchased from VWR . Dopamine hydrochloride (99%) was obtained from Fisher Scientific . In-house DI water was further purified using a Milli-Q Reference system. Tris(hydroxymethyl)aminomethane  was obtained from Sigma Aldrich. Adhesive-backed polyethylene sheets  and stainless steel 301 were purchased from McMaster-Carr , washed with (70%) ethanol, rinsed with DI water, and dried with nitrogen gas. Cu2O coating was applied as described previously [2O in ethanol was subsequently applied on the PU film, and it was left to partially dry for a few min. The coated wrap was then heat-treated in an oven (120 °C for 2 h) to speed up the formation of the PU coating, washed using DI water, and cut into smaller pieces (12 × 12 mm). In some cases, where indicated, the wrap was plasma-treated with Argon . The antimicrobial wrap was applied to stainless steel.A PU/Cueviously . In brie2O wrap was prepared using the method in reference [2O particles (0.2% w/w) in 10 mm tris solution was sonicated, and dopamine hydrochloride was added to the suspension to yield a final concentration of 0.05 g dopamine/L. Subsequently, 140 µL of the prepared suspension was applied to the uncoated wrap (15 × 15 mm in size). The sample was heat-treated in an oven (80 °C for 30 min). The antimicrobial wrap was applied to stainless steel.The PDA/Cueference . A suspeThe morphology of the coatings was characterized via scanning electron microscopy (SEM) using either a JEOL IT500 SEM  or a FEI Quanta 600 FE-ESEM . Samples for SEM were treated with a 5 nm layer of Pt/Pd to enhance electrical conductivity.P. aeruginosa (strain DSM-9644) was chosen to test the antimicrobial properties of the samples.According to the United States Environmental Protection Agency ,42 guidaP. aeruginosa were confirmed before using the batch to test the antimicrobial coatings.Bacteria were grown to the mid-exponential phase at 37 °C in 5 mL of Tryptic Soy Broth (TSB) with aeration (rpm = 60). For the visual verification of the identity and purity of the bacterium, cells were streaked from the liquid culture on Tryptic Soy Agar (TSA) plates and then incubated (37 °C for 48 h). The specific color and morphology traits of g), followed by the removal of the supernatant and then re-suspension in 5 mL of Phosphate-Buffered Saline (PBS), followed by vortexing (60 s). Two rounds of centrifugation and resuspension in PBS were carried out.Bacterial cells were extracted from liquid cultures via centrifugation . CFU/mL of the suspension was measured by spreading 0.1 mL of a suspension on a TSA plate (in triplicate), and colonies were counted after incubation for 48 h at 37 °C. Because of the large range of CFU, each test suspension was used to create a 10-fold dilution series, and each of these dilutions were measured to obtain the optimum number of CFUs for counting (20–200). The suspensions had a CFU/mL of approximately 1.6 × 10P. aeruginosa cells in PBS was placed on solid test samples six days after the antimicrobial wraps were prepared. An uncoated wrap was used as the control. After a known time interval , an individual sample was placed in a 50 mL centrifuge tube containing 5 mL of PBS. The sample was then vortexed (10 s and at the highest setting) and sonicated  to detach all cells from the solid. Assuming the resulting suspension of cells was homogeneous, the total number of surviving cells could be measured by obtaining the CFU/mL of this bacterial suspension. In total, 0.1 mL of the suspension was removed without contacting the sample, and the CFUs were measured as described above. In addition, the CFU/mL for input bacterial suspension was measured for the calculation of survival. Three independent solid samples were tested in each condition , and three repeats were carried out for each of these independent measurements. The detection limit and resolution on the agar plate was 1 colony, but because only 0.1 mL of the 5 mL volume was sampled, the detection limit of bacteria was 50 CFU or log (CFU) = 1.7.A 5 µL droplet of t-tests were two-tailed and heteroscedastic. A significance level of 0.05 was employed for statistical analysis. The half-life was calculated from a linear fit of the slope of all log (CFU)–time data at the three time points. The half-life is:All statistics were calculated from log CFUs, and Student’s All data from 0 to 20 min were used to calculate The CFU counting technique counts cells that can reproduce enough times to form a visible bacterial colony. We use the term “kill” as a shorthand for cells that cannot form a colony. Percentage kill is a comparison of the CFUs measured after a specific time to the CFUs of the input suspension:For calculations, the measured sample CFUs were adjusted for dilutions that took place during extraction of bacteria from the solid. When compared to a standard at the same time, we used reduction, defined as:p-values for 99.9% reduction were obtained by the following method: for each of the three independent measurements, we calculated:t-test to determine whether those values were different to the three independent values of p-value indicated the confidence that the antimicrobial coating decreased the cell numbers by more than 99.9%.We began the statistical treatment with a log transformation because we found that the residuals were approximately normally distributed after a log transformation. Each CFU measurement in Equations (3) and (5) is the mean of three repeat counts of a different agar plate from the liquid sample; each mean in Equations (3) and (5) is the mean from three measurements on three independent samples.


Following viral infection, viral antigens bind specifically to receptors on the surface of lymphocytes thereby activating adaptive immunity in the host. An epitope, the smallest structural and functional unit of an antigen, binds specifically to an antibody or antigen receptor, to serve as key sites for the activation of adaptive immunity. The complexity and diverse range of epitopes are essential to study and map for the diagnosis of disease, the design of vaccines and for immunotherapy. Mapping the location of these specific epitopes has become a hot topic in immunology and immune therapy. Recently, epitope mapping techniques have evolved to become multiplexed, with the advent of high-throughput sequencing and techniques such as bacteriophage-display libraries and deep mutational scanning. Here, we briefly introduce the principles, advantages, and disadvantages of the latest epitope mapping techniques with examples for viral antigen discovery. Adaptive immunity plays a vital role in the elimination of pathogens and the protection of organisms from re-infection. Adaptive immunity relies primarily on two types of lymphocytes, B cells and T cells, which mature in the bone marrow and thymus respectively and enter the peripheral lymphoid organs via the circulatory system . When foThese foreign poly-peptides that stimulate adaptive immunity are collectively referred to as antigens. There are many different pathogens in nature, which in turn contain a variety of antigens. An effective antigen should have two main functions: immunogenicity, the ability of the antigen to efficiently activate the proliferation and differentiation of naive lymphocytes; antigenicity, the ability of the antigen to bind with high specificity to the immune effector cell . As anti26 antibodies in different sequence combinations , and differentiate into plasma cells and memory B cells with the same antigenic specificity. Plasma cells secrete antibodies that bind specifically to antigens and are important effector molecules in mediating humoral immunity against pathogens. The structure of an antibody is closely linked to its function. Antibody structures can be divided into variable region (V region) and constant region (C region) . The reginations , 14. Wheinations . Plasma inations , 17. Antinations . At highinations .The antigen recognition receptors on the surface of T cells are also membrane surface immunoglobulins and are known as T cell receptors (TCRs). Similar to BCRs, TCRs are also divided into V regions and C regions, and therefore TCRs have the ability to recognize, bind to and deliver activation signals to the cell . TCRs caBoth B cells and T cells have the capacity to recognize various epitopes that suit a variety of functions. Epitopes may be further classified into the following classifications: linear epitopes and conformational epitopes, based on their structural distinctions. Linear epitopes, also known as continuous epitopes, consist of amino acid residues arranged in a continuous sequence. Conformational epitopes, also known as a discontinuous epitope, consists of amino acid residues that are not continuously arranged but are close to each other in spatial structure . While BDuring infection, diverse viral epitopes stimulate the host to produce a wide variety of antibodies and cytotoxic T cells , 27. AntEpitope mapping technology is essential to clarify the natural antigenic landscape and evaluate the most potent and effective epitopes for an appropriate immune response. Common B cell epitope mapping techniques include Deep Mutational Scanning, peptide/protein microarrays, bacteriophage peptide/protein display and other peptide display techniques. Common T cell epitope mapping techniques are divided into three categories, peptide-MHC multimer-based, cell-based and yeast-based epitope mapping techniques. Here, we cover the latest in multiplexed epitope mapping technology Table 1 Traditional immunology has relied on specific antibodies generated against a single, well-defined, conformational epitope or a short linear section of a protein (often <50 aa’s). While whole proteins are sometimes utilized to generate antibodies, manual mapping of the antibody epitope must then occur through a range of somewhat arduous techniques. These techniques may provide high specificity and detail but they lack the capacity for en-bulk high-throughput mapping.Deep Mutational Scanning (DMS) utilizes gene mutagenesis, coupled with other techniques such as high-throughput sequencing, to obtain a saturated library of mutations covering antigenic epitopes , 35. DMSIn 1984, Geysen et al. first proposed a technique for the rapid synthesis of peptides on the surface of immobilized carriers, followed by screening the immobilized peptides for antigenic epitopes using antibodies. Subsequently, Frank and Fodor et al. proposed new peptide synthesis techniques, SPOT synthesis and photolithography synthesis, respectively , 42. ThePeptide/protein microarrays can be used to characterize antibody responses. Gallerano et al. prepared a set of microarrays covering HIV-1 clade C proteins and peptides that can be used to diagnose HIV and detect changes in specific antibody responses as HIV infection and treatment progress . ValentiThe Sengenics KREX chip is a commercially available protein microarray that immobilizes proteins with full-length sequences, correctly folded carrier and functional validation. Due to the presence of biotin carboxyl carrier protein (BCCP), BCCP-tagged biotinylated KREX proteins can only be immobilized on the chip if the carrier protein is correctly folded . The majThe identification of important immunogenic epitopes of pathogens plays a key role in the development of targeted antibody therapies. Gu et al. used peptide arrays to screen the epitopes recognized by antibodies in the sera of patients at different stages of hepatitis B infection . They foDue to their flexibility, speed, relative affordability and high throughput, peptide/protein microarrays are widely used in the development of new diagnostics, therapeutic antibodies, and vaccines against pathogens. However, there are still many shortcomings with the automated use of peptide microarrays. Current techniques for constructing novel peptide/protein microarrays are costly and inefficient –55. When11 pfu/ml in size and can contain thousands of different peptides/proteins  for identifying antigenic epitopes recognized by antibodies in autoimmune diseases . Unlike Phage-display libraries may be used to quickly screen multiple different peptide or protein display libraries. One particular PhIP-seq library, Virscan, encodes a peptide display library covering the antigenic epitopes of the currently known human viral proteome . Due to 11), phage display techniques utilizing synthetically produced libraries still have an irreplaceable advantage  . While ldvantage , 64, 72.Bacterial surface display is a common technique for displaying epitopes on the surface of bacteria. First, a suitable host is selected as the display platform . Gram-ne8-109, significantly less than other peptide display techniques . This limitation has been partially ameliorated with integration of CRISPR-Cas9, or another approach that utilizes libraries from immunized animals with initial antibody screening and maturation in vivo  complex by an APC in order to be specifically recognized by the TCR. Overlapping peptide pools have previously been used for systematic screening and epitope identification of Infectious Bronchitis Virus in humans by direct stimulation in PBMCs, using readouts such as ELISpot or cytokine production measured by flow cytometry , 100. ThAs this complex heteromeric MHC protein is normally folded inside the cell prior to loading a peptide , this key p-MHC complex formation is essential for T cell epitope mapping. The generation of libraries for T cell binding therefore goes well beyond just the generation of antigens. In the absence of co-stimulation, p-MHC has a very weak affinity for TCR(1-100 μM), dissociating rapidly and making it difficult to detect the interaction between TCR/p-HMC . Alman eWhen cells come in contact, the phenomenon of sharing and transferring membranes and membrane-associated proteins between cells is called trogocytosis . When T 6 epitopes, but significantly less than the diversity of the yeast display library. False positive results have occurred in some cases and further refinement of the technique is required to improve the specificity of the assay.In addition, they have developed another epitope mapping technique that uses a chimeric receptor called the signal transducer and antigen presenting antigen bifunctional receptor (SABR) to identify TCR-p-MHC interactions . SABR, aUsing both approaches, homologous epitopes of novel TCRs isolated from melanoma patients were successfully retrieved. The feasibility of these two cell-based epitope mapping techniques in exploring homologous epitopes of TCRs was demonstrated, to guide the site of action of immunotherapy , 113. ThT-scan is a genome-wide approach that allows comprehensive scanning and discovery of cytotoxic T cell epitopes . The keyAs described earlier for B cell epitopes, the structure of proteins displayed through yeast is highly similar to that of mammals, and therefore yeast display can be applied for high-throughput epitope mapping of T cells. Yeast libraries are specifically generated with a high diversity of peptides though unlike B cell epitope mapping, the display libraries utilize a fusion protein with a cell wall anchoring protein expressed on the cell surface. This gives each yeast the ability to display a unique p-MHC on its surface . The yeaEscherichia coli, suggesting that the presence of antigenic cross-reactivity between tumor and virus may be a novel feature in oncogenesis  epitope, TMEM161A, overexpressed in cross-reactive epitopes of Epstein-Barr virus and ogenesis .In contrast, the yeast agglutination-mediated TCR antigen discovery system (YAMTAD) enables rapid and efficient confirmation of p-MHC-TCR interactions even in the absence of TCR tetramers with the aid of yeast display and engineered yeast mating . The basOther techniques for T cell epitope identification have also been developed recently, including lentiviral expression libraries containing antigens for binding of TCRs and BCRs, coupled to scRNA-Seq for simultaneous identification of both antigen (post-integration into the genome) and the TCR/BCR itself from the same cell . This tePredictive structural modelling of proteins with the likes of AlphaFold has dramatically improved accessibility of computationally predicted structures. However, epitopes and conformational epitopes, are often flexible regions that do poorly with structural prediction due to the lack of constraints. Additionally, modelling and docking of peptides inside the MHC-groove and between MHC, peptide and TCR are still relatively poor predictors of true binding dynamics. Recent advances in machine-learning driven TCR-epitope binding are improving, however, structure-based epitope mapping techniques are still an indispensable tool for understanding many epitopes . While tX-ray crystallography not only enables the construction of 3D structural models of antigen-antibody complexes, but also the detailed description of epitope, paratope and the interaction forces between them. Koenig et al. designed four SARS-CoV-2 neutralizing nanobodies and identified two different epitopes by x-ray crystallography, among which one bound by VHH E overlaps highly with ACE2 . HoweverNeisseria meningitidis vaccine and thus guide the improvement of vaccines . Hydrogen/deuterium exchange, in the core binding region of antibody-bound epitope, is significantly lower than in the unbound region, allowing a readout for specific information on conformational epitopes . Mass spvaccines . Combinevaccines , 142. Mavaccines , 144.10-1014) coupled with classical structure-based techniques for fine conformational identification. T cell epitopes require the design of peptide libraries based on specific MHC molecules which poses difficulties due to the high polymorphisms of MHC molecules and more complex binding dynamics. Still, many tools for T cell epitope mapping have been utilized, although on a much smaller scale than those for B cells, to obtain high-resolution epitope information. These techniques, summarized in Adaptive immunity generates long-term immunological memory and can stimulate powerful pathogen clearance; as such, antigenic epitopes play an important role in immunotherapy and vaccinology as a ‘switch’ to stimulate initiation. Epitopes associated with B cells can generally be characterized by the specific binding to antibodies generated by B cells. T cell epitopes are generally considered to be more simple, smaller epitopes compared to antibody binding, but their recognition is tightly associated with MHC loading and presentation. Both are associated with high-specificity and a great diversity of potential epitopes. This in turn increases difficulty for identification of specific epitopes and leads to several differences in epitope mapping techniques between B and T cells. Despite this, many mapping tools have been developed, allowing high-throughput epitope analysis  becomes cheaper and more high-throughput, it facilitates more comprehensive characterization of higher diversity libraries . DMS, inCurrent techniques for epitope mapping have evolved towards large scale epitope screening, combined with lower cost, higher sensitivity, and increased specificity. Advances in sequencing, scRNA-Seq, barcoding, surface display libraries, DMS and cell-free binding have all rapidly expanded our knowledge of epitopes. Further utilizing this high-throughput data with machine-learning algorithms may also facilitate optimal epitope prediction methods, with higher success rates than what is currently observed, to advance targeted vaccine design. This will also help to explore the general immune response process during viral disease, autoimmune disease, tumor progression and provide future guidance for disease diagnosis, vaccine development and immunotherapy –149. TheHD & AI wrote and composed the manuscript, sourced references and summarized appropriately. AI conceptualized the idea. Both authors contributed to the article and approved the submitted version.


Increases in food production to meet global food requirements lead to an increase in the demand for nitrogen (N) fertilizers, especially urea, for soil productivity, crop yield, and food security improvement. To achieve a high yield of food crops, the excessive use of urea has resulted in low urea-N use efficiency and environmental pollution. One promising alternative to increase urea-N use efficiency, improve soil N availability, and lessen the potential environmental effects of the excessive use of urea is to encapsulate urea granules with appropriate coating materials to synchronize the N release with crop assimilation. Chemical additives, such as sulfur-based coatings, mineral-based coatings, and several polymers with different action principles, have been explored and used for coating the urea granule. However, their high material cost, limited resources, and adverse effects on the soil ecosystem limit the widespread application of urea coated with these materials. This paper documents a review of issues related to the materials used for urea coating and the potential of natural polymers, such as rejected sago starch, as a coating material for urea encapsulation. The aim of the review is to unravel an understanding of the potential of rejected sago starch as a coating material for the slow release of N from urea. Rejected sago starch from sago flour processing is a natural polymer that could be used to coat urea because the starch enables a gradual, water-driven mechanism of N release from the urea–polymer interface to the polymer–soil interface. The advantages of rejected sago starch for urea encapsulation over other polymers are that rejected sago starch is one of the most abundant polysaccharide polymers, the cheapest biopolymer, and is fully biodegradable, renewable, and environmentally friendly. This review provides information on the potential of rejected sago starch as a coating material, the advantages of using rejected sago starch as coating material over other polymer materials, a simple coating method, and the mechanisms of N release from urea coated with rejected sago starch. In plants, nitrogen (N) deficiency leads to symptoms such as stunted growth, yellowish leaves (chlorosis), and a thin and spindly stem . Nitroge3−) leaching, ammonia (NH3) volatilization, and denitrification [3− into the groundwater and hindering the volatilization of NH3 into the atmosphere. In another related study on encapsulated urea, Mulder et al. [Changing the rate of urea dissolution in the soil to synchronize the availability of inorganic N with crop requirements could be achieved by coating the urea to ascertain an improvement in the NUE. Moreover, owing to the fast release of N from the urea and the N becoming readily available in the soil, not all the N is used by plants due to factors such as the plants not efficiently absorbing 40 to 70% of the N applied from urea, which results in a low N uptake and NUE [fication ,6. The sfication . Accordification , the encr et al.  revealedr et al. .The advantages of coated urea over conventional urea are cost savings and a reduced application frequency in the case of short-term crops, such as leafy vegetables, because less urea is applied to sufficiently provide the N required for the growth season when N is available in the soil for crop uptake . Coated 3− leaching, and N loss, as indicated in The materials used for coating urea can be divided into organic and inorganic coatings, as summarized in Starch, a naturally based coating material, is the most-researched natural polymer for coating urea due to its high availability, low cost, and environmentally friendly nature . RychterCompared with the uncoated urea granules, which released N into water in 6 min under mild shaking, the corn-starch-coated urea took approximately 32 min to completely release N . In this review, focus is narrowed to the use of organic materials for urea coating, as summarized in The use of a natural, biodegradable polymer such as starch for urea coating constantly increases because when starch-coated urea is applied to the soil, the degradation begins with the presence of bacteria, fungi, and algae enzymes which catalyze the process of chemical hydrolysis . In addiSome of the methods adopted for urea coating are pan coater, rotary drum, and fluidized bed coater technology . Among t3 volatilization from urea [For the rotary pan operation procedures, urea particles are shaken by the rotation of the pan and sprinkled with solution by the spray nozzles. Throughout the sprinkling operation, high-temperature air is applied over the wet coatings for drying purposes . When throm urea . In our rom urea . To comprom urea . Apart fw/v of starch, carboxymethyl cellulose (CMC), and 1 g of polyethylene glycol (PEG) [3 loss and NO3− leaching from the application of urea. Our study confirmed that the lower dissolution rate of coated urea using an RSS concentration of 2% was found to be suitable for encapsulating urea compared with other concentrations of 3%, 4%, 5%, and 6%, respectively. The lower dissolution rate with the lower RSS concentration used for the urea coating material in our study contradicted the findings of Naz and Sulaiman [In our previous work, we coated urea using a self-built mini rotary machine which was assembled from two stainless steel basins with a diameter of 30 cm and a depth of 10 cm that were attached to a driller that rotated the basins. A fan regulator was used to control the speed of the rotary machine. In our study, the method of preparing the coating materials was slightly modified from the method of Himmah et al. . Himmah ol (PEG) , whereasSulaiman . IncreasSulaiman . The economic activity of sago starch flour production in Sarawak, a state in East Malaysia, has played an essential role in sustaining rural livelihood, either for subsistence, trading at the local and domestic market, or for export to Taiwan, Japan, China, and Korea. As shown in The sago segments are commonly immediately transported to the mill for sago flour processing to ensure the premium quality of the sago starch and to avoid deterioration of the starch in the trunk. Sago palms are harvested for starch at the age of 7–15 years, when the starch fully developed in the sago stem and before flowering . The ste4+. Additionally, RSS has a high number of basic cations, especially K+ (170.27 cmol(+) kg−1) and Mg2+ (20.3 cmol(+) kg−1), and its inherent basic cation content helps to enhance its CEC, suggesting an improvement in soil fertility and nutrient retention [In our previous study on coating urea granules with RSS, we analyzed the selected physico-chemical properties of RSS, as summarized in etention . Both thetention . Sago st4+), which is then taken up by the plants. In addition, micropores allow small molecules such as NH4+ and NO3− to diffuse through the hydrogel [4+ and NO3−) in a delayed manner that is in synchrony with the needs of crops for NH4+ and NO3−, thus improving the N use efficiency with an enhanced yield. As sago starch is a natural, hydrophilic polymer, the movement of N from the polymer into the surrounding soil is driven by a mass transfer resulting from a concentration gradient and by Brownian molecular motion (self-diffusion). In this way, the diffusion of nutrients from the polymer is similar to that of the diffusion from other inorganic sorbents used as release-controlling devices. Thus, the result of the restricted diffusion of nutrients from within the polymer can delay the release of nutrients into the surrounding soil. Moreover, the unique properties of sago starch, such as hydrophilic nature, enable it to dissolve in water. Although sago starch is hydrophilic, it is highly water sensitive, which can prevent water from entering into the starch film, thus slowing down the hydrolysis of urea by preventing water from reacting with the urea granule. Hydrophilic polymers are characterized by the presence of a main polymer “backbone” with attached polar groups, and they contain micropores. The presence of abundant hydrophilic groups (sago starch contains a hydroxyl group) and micropores in the starch results in the protective layer being less resistant to water and easier to crack, enabling the dissolution of the nutrients. The presence of functional groups can help to retain a positively charged ion  and preventing anion–anion electrostatic repulsion in the network, decreasing the swelling capacity. In contrast, the presence of Na+ ions in the solution shields the COO− anion and prevents anion–anion electrostatic repulsion in an alkaline environment (>pH 9). Thus, the pH of the coating material is important, as it can acidify the soil microsite and reduce the swelling capacity. In our study, coating urea with RSS minimized the emission of NH3 because the acidity of RSS, which is 3.43, enables the urea granule surface to be temporarily acidified. In addition, when the pH is less than 5.5, urea hydrolyzes slowly [The morphology of uncoated urea and RSS-coated urea were analyzed under scanning electron microscope (SEM), which showed that the uncoated urea granule surface was rough, with fine openings and a porous structure through which water can penetrate to dissolve the urea granule. On the other hand, a decrease in membrane porosity and uniform layer were observed in surface morphology of the RSS-coated urea . The RSSs slowly ,77. ConsThe mechanism of the slow release of N from coated urea reflects the effectiveness of a slow-release fertilizer and the fertilizer’s efficiency. However, the slow release of N from coated urea is complicated to visualize because the release mechanism of N from coated urea depends on several factors, such as the nature of the coating material, atmospheric temperature, soil moisture, agricultural environment, farming methods, and many other variables. As summarized in 4+ in the soil, 4+ for plant uptake, leading to increase in the NUE and minimizing the frequency of N fertilizer application. In addition, the coated urea mitigates N loss through the volatilization of NH3 and the emission of nitrous oxide (N2O) in such a way that the coating serves as a nitrification inhibitor, which reduces the rate of nitrification of NH4+ to NO3−. Thus, N2O and nitric oxide (NO) emissions, NO3− leaching and/ or run-off, and the denitrification of NO3− are reduced during this process. The mechanisms of N release from the coated urea take place in three different stages, such as the lag, linear, and decay regions, while the uncoated urea follows a linear trend  against losses through mineralization. In the early stage of incubation, uncoated urea dissolved rapidly due to its high solubility and the rapid urease-catalyzed hydrolysis of urea, whereas RSS-coated urea demonstrates a delayed solubility due to the presence of the physical barrier of the coating layer; thus, N loss is lower than for the uncoated urea (4+. As a result, the RSS-coated urea retained N in the soil when compared with the uncoated urea (ted urea . Most ofted urea . At 80 dted urea . Compareted urea . The sloAlthough natural materials are renewable, biodegradable, and environmentally friendly, some organic coating materials have disadvantages, as summarized in The production of slow-release fertilizers with biodegradable coatings, such as the use of RSS, is a huge contribution to soil ecosystem and organic farming. To achieve fully organic farming, other native starches, such as potato starch, were used as a urea coating. A single layer of native potato starch substance for urea coating increased the static strength of the urea granules by up to 22.7 N/granule, while a two-layered coating reduced the granules’ strength to 19.3 N/granule but was still higher than the minimum requirement provided by fertilizer producers, which is 7.0 N/granule . The mecTo that end, this paper reviewed the potential of RSS, the solid waste from sago flour processing, as a coating material for urea encapsulation. The proper application and better understanding of the function of RSS as coating material could improve the retention of soil-available inorganic N by controlling the rapid release of N during urea hydrolysis due to the specific physicochemical properties of the RSS. Inorganic, organic, and natural polymers have been widely explored for the formulation of urea encapsulation with the properties of slow-release fertilizers or controlled-release fertilizers. In addition to its function as a natural polymer for urea encapsulation, starch is also used as a binder and filler. Starches that have been modified for their thermophysical and mechanical strength are very important in the release pattern because each starch has specific properties that can facilitate the desorption of N from the coated urea. The existing materials for urea coating are not necessarily the best options because they have some drawbacks, such as being expensive and non-biodegradable. This review revealed the potential of RSS as a urea coating material for agricultural means and highlighted the benefits of RSS, which is abundantly available, low-cost, and considered waste in sago starch flour processing. In terms of agronomic efficiency, the coating method is easy and does not require any expensive equipment. Thus, the adoption of the RSS-coated urea by farmers could be affordable. From economic and environmental efficiency perspectives, the use of RSS is considered low-cost because RSS is waste generated from sago starch processing that has the potential to pollute the environment.