Neuronal intranuclear inclusion disease (NIID) is a rare neurodegenerative condition characterized by the loss of neurons and the presence of eosinophilic nuclear inclusions in the central and peripheral nervous system, skin and visceral organs. In this paper, we present a case of NIID with recurrent encephalitic attacks that remained stable and nonprogressive for seven years; no such case has previously been reported.NOTCH2NLC  gene confirmed the diagnosis of NIID. She received supportive treatment such as nutrition support therapy and vitamin B and C supplementation, as well as symptomatic treatment during hospitalization. The patient’s symptoms were completely relieved within one week.A 63-year-old female was hospitalized due to light-headedness, vomiting, unstable gait and cognitive impairment. Seven years prior, she had experienced an episode of light-headedness, central facial paralysis, unstable gait, aphasia, nausea, vomiting and loss of consciousness. She regained consciousness within 12 h, and her other symptoms were completely resolved within one week. During the present hospitalization, a brain magnetic resonance imaging (MRI) examination detected high signal intensity on diffusion-weighted imaging (DWI) of the bilateral frontal grey matter–white matter junction. We reviewed the patient’s previous MRI results and found that she had also had high signal intensity on DWI of the bilateral frontal grey matter–white matter junction seven years prior. In the intervening seven years, the high signal intensity in the frontal lobes had spread along the grey matter–white matter junction, but the deep white matter remained unaffected. Skin biopsy was performed, and intranuclear inclusions were found in adipocytes, fibroblasts and sweat gland cells. GGC repeat expansions in the This is a detailed report of a case of NIID with multiple reversible encephalitic attacks, diagnosed by clinical symptoms, intranuclear inclusions, characteristic DWI signals, and genetic tests. NOTCH2NLC  gene is associated with the mechanism of NIID [NIID is a rare, slowly progressing neurodegenerative disease characterized by eosinophilic intranuclear hyaline inclusions in cells of the central and peripheral nervous system (including the autonomic nervous system) and the visceral organs . GGC rep of NIID . Dementi of NIID . While a of NIID . Dementi of NIID . Diagnos of NIID .We report a case of NIID with multiple reversible encephalitic attacks, in which the patient experienced two encephalitic episodes followed by complete remission of symptoms. As illustrated by this case, clinicians need to be alert to the possibility of NIID when patients have reversible encephalitic attacks.9/l), and a normal leukocyte count (3 × 106 monocytes/l). The patient’s nerve conduction studies were normal. An electroencephalogram (EEG) showed no epileptic discharge. A review of the patient’s brain MRI results from 2011 revealed the presence of a high signal intensity on DWI of the bilateral frontal grey matter–white matter junction  premutation.A 63-year-old woman was admitted to hospital with an acute neurological episode. The patient’s complaints were light-headedness, vomiting, unstable gait and cognitive impairment. She had been healthy except for a similar attack seven years prior and had no family history of similar symptoms. Upon admission, the patient’s body temperature was 36.7 °C, and her blood pressure was 122/86 mmHg. Neurological examination revealed no abnormalities in cranial nerve function or muscle strength. The bilateral knee reflex and Achilles tendon reflex were diminished. The biceps and triceps reflexes were normal. The bilateral plantar responses were flexor. Her gait was wide-based and ataxic. She did not have increased muscle tone or bradykinesia. Her complete blood count was normal, as were her liver and kidney function. Neuropsychological examinations revealed mild cognitive impairment: 24/30 on the Mini-Mental State Examination (MMSE), 25/30 on the Montreal Cognitive Assessment (MoCA), 10 on the Hamilton Depression Scale (HAMD), 24 on the Hamilton Anxiety Scale (HAMA), 3 on the Neuropsychiatric Inventory (NPI) and 33 on the Scales for Outcomes in Parkinson’s Disease–Autonomic (SCOPA-AUT). The decreases in the MMSE and MoCA scores were caused mainly by reductions in the patient’s short-term memory and computational ability. Brain MRI revealed high signal intensity in the bilateral frontal grey matter–white matter junction on DWI . The expansion of repeats in the 5′ UTR of the NOTCH2NLC gene confirmed the diagnosis of NIID. At the time of this report, the patient’s outcome is good.In this case study, the patient had two acute encephalitic attacks that were manifested mainly as loss of consciousness, neuropsychological impairments, gait disorders, autonomic dysfunction, etc. Normal body temperature, complete blood count, and cerebrospinal fluid cell counts helped exclude infectious encephalitis. The characteristic high signal intensity on DWI alerted the physicians to perform skin biopsy and genetic testing. The skin biopsy revealed intranuclear inclusions in adipocytes, fibroblasts and sweat gland cells; this set of features was compatible with NIID. A negative test result for the FMR1 gene premutation and expansion of repeats in the NOTCH2NLC gene.This case study describes a patient with NIID who presented with two acute reversible encephalitic episodes over seven years. The patient was diagnosed on the basis of characteristic DWI signals, intranuclear inclusions, a negative NOTCH2NLC and the absence of FMR1 gene premutation, she was diagnosed with sporadic adult-onset NIID [FMR1 genetic testing, which showed that the number of CGG repeats was within the normal range (< 44 repeats).In light of the patient’s GGC repeat expansion within human-specific set NIID . We exclOver the seven-year period of disease progression, high DWI signal spread along the grey matter–white matter junction, but the deep white matter was not affected. The signal abnormality progressed continuously once it appeared, which is consistent with previous reports .Due to the highly variable nature of the clinical symptoms, NIID is considered a heterogeneous disease . Most ofNIID is characterized pathologically by the presence of eosinophilic intranuclear inclusions in neuronal and glial cells , 15. FacNOTCH2NLC gene alterations [NOTCH2NLC is highly expressed in various populations of radial glia and is associated with evolutionary expansion of the human cerebral cortex [NOTCH2NLC lacks the N-terminal signal peptide and has a 2-bp deletion just downstream of the NOTCH2 start codon. Moreover, by long-read sequencing, GGC repeat expansion in the 5′ region of NOTCH2NLC was identified in all patients with familial NIID as well as those with sporadic NIID. Sone et al. found that the change in the number of repeats ranged from 6 to 61 in control subjects and 71 to 183 in NIID patients [NOTCH2NLC gene is the main cause of NIID and related diseases [Recently, several studies suggested that NIID was associated with erations , 22. NOTl cortex . NOTCH2NIn summary, we present a case of NIID with multiple reversible encephalitic attacks; such a case has never been previously reported. The case was diagnosed by intranuclear inclusions on skin biopsy and by genetic testing; these diagnostics were carried out in response to high-intensity DWI signal at the grey matter–white matter junction. Clinicians need to be alert to the possibility of NIID when patients exhibit reversible encephalitic attacks and intense DWI signal at the grey matter–white matter junction.


Starches were extracted from chickpea (C.P.), corn (C.S.), Turkish bean (T.B.), sweet potato (S.P.S.), and wheat starches (W.S.). These starches exhibited different amylose contents. The extracted starches were annealed in excess water and in germinated sorghum extract (GSE) (1.0 g starch/9 mL water). The α-amylase concentration in the GSE was 5.0 mg/10 mL. Annealing was done at 40, 50, and 60 °C for 30 or 60 min. The pasting properties of annealed starches were studied using Rapid Visco-Analyzer (RVA), in addition to the swelling power. These starches exhibited diverse pasting properties as evidenced by increased peak viscosity with annealing, where native starches exhibited peak viscosity as: 2828, 2438, 1943, 2250, and 4601 cP for the C.P., C.S., T.B., W.S., and S.P.S., respectively, which increased to 3580, 2482, 2504, 2514, and 4787 cP, respectively. High amylose content did not play a major role on the pasting properties of the tested starches because sweet potato starch (S.P.S.) (22.4% amylose) exhibited the highest viscosity, whereas wheat starch (W.S.) (25% amylose) had the least. Therefore, the dual effects of granule structure and packing density, especially in the amorphous region, are determinant factors of the enzymatic digestion rate and product. Swelling power was found to be a valuable predictive tool of amylose content and pasting characteristics of the tested starches. The studied starches varied in their digestibility and displayed structural differences in the course of α-amylase digestion. Based on these findings, W.S. was designated the most susceptible among the starches and S.P.S. was the least. The most starch gel setback was observed for the legume starches, chickpeas, and Turkish beans (C.P. 2553 cP and T.B. 1172 cP). These results were discussed with regard to the underlying principles of swelling tests and pasting behavior of the tested starches. Therefore, GSE is an effortless economic technique that can be used for starch digestion (modification) at industrial scale. Starch molecular structure contrasts by botanical source. The X-ray patterns of starches such as corn and wheat have an A-type crystal within the granule, while tubers exhibit B-type. Unlike the B-type, the A-type crystal is a closely packed arrangement of double helices which is more stable thermodynamically. The temperature during the growth of the plant could influence starch composition (ratio of amylose to amylopectin) and structure . EndotheGranular (native) starch digestion (hydrolysis) is not as fast as gelatinized starch whose crystallinity has been essentially damaged; amylose and amylopectin became more accessible to enzymes and not restricted by the double helices or amylose-lipid complexes. Bacteria, fungi, plants, and human α-amylase are common endo-enzymes known to hydrolyze α-(1-4) bonds in a random fashion, thereby decreasing the molecular weight of starch molecules. Previous studies have shown that the action of human α-amylase on starches of different botanical origin occurred in various digestion kinetics pathways and produced diverse degradation products .The characteristics of native starch granules that regulate the location, rate, and extent of hydrolysis by α-amylase are not yet known. Nonetheless, the size and shape of granules are controlling factors. Access of α–amylase to its substrate and the release of reaction products are influenced by many factors such as granule integrity, crystallinity, and porosity, amylose to amylopectin ratio, structural, phosphate content (in tubers), lipids on the surface of starch granules, and starch degradation products such as dextrin that have been shown to inhibit α–amylase . TherefoThe factors that contribute to the differences in the kinetics of α-amylase interaction with starch granules are still not well defined. In addition, the changes that occur in starch granules structure during heat processing of foods make starch digestion enzymology puzzling. The variations of α-amylase interaction with starch are mainly accredited to inherent distinctions in the granule structure of diverse botanical species in addition to food processing ,8.At small scale, we may picture the surface of a starch granule to differ significantly between starch granules according to origin, from fairly smooth, assembled from small blocklets fixed on an amorphous matrix, to a rougher surface rising from large crystalline blocklets extended from an amorphous structure ,10. It iThe objectives of this work were to determine the effect of crude extract of germinated sorghum seeds on the physicochemical, pasting, and swelling power of wheat, chickpea, sweet potato, corn, and Turkish beans starches. This study includes the effect of annealing on the physical and pasting properties of the isolated starches. Pure α-amylase enzyme was used parallel to the crude extract so that to assist in monitoring the extract effectiveness. The outcome of this work will allow for using economical crude (un-purified) α-amylase extract in the starch industry for starch physical modification or syrup production.Aspergillus fungal α-amylase (EC3.2.1.1) were purchased from Sigma-Aldrich . The centrifugation step of starch isolation was done using Beckman Centrifuge .The raw material was purchased from a local supermarket in Riyadh (Saudi Arabia). The sweet potato used was Salem variety grown in Altayef area of the western Saudi Arabia in the winter, whereas wheat was a hard red winter wheat grown in the north part of Saudi Arabia, the Hail region, in the winter. The chickpea and Turkish bean were purchased from the local market . Corn starch was donated by ARASCO Company . Analytical grade sodium hydroxide, acetic acid, and weight/volume) in a heavy-duty blender for 5 min  and passed through 200 mesh sieves and centrifuged at 2000× g for 15 min at 10 °C. The top layer on the precipitate was removed and the white substance at the bottom of the bottle was suspended in distilled water and centrifuged using the above-mentioned conditions. This procedure was repeated five times to obtain pure white starch  for 3 min using an auxiliary kitchen mixer . The slurry was filtered through a muslin cloth and the overs were re-blended and filtered in the same way. The starch in the filtrate is allowed to settle for one hour at room temperature and the top liquid was discarded. The precipitate was re-suspended in distilled water and centrifuged at 2000×g for 15 min at 10 °C. After centrifugation, the top layer was removed and the white matter at the bottom of the bottle was reconstituted in distilled water and centrifuged three times until pure white starch was obtained. The isolated starch was air-dried and stored in airtight jars at 5 °C.According to the method of Sit et al. , sweet pAmylose content of the isolated starches was determined according to the method of Williams et al. . The met×g. Fresh extract was prepared daily.Germination sorghum was prepared according to Afify et al.  with minw/v). To the slurry, 0.1 mL and 1.0 mL of extract were added and one set of samples was annealed without GSE. The slurry was placed in a shaking water bath at 40, 50, and 60 °C for 30 or 60 min with constant agitation at 100 rpm . The digested slurry was centrifuged to remove the digestion product, the supernatant was discarded, and the precipitate was air-dried. The centrifugation process was repeated three times by adding water to the precipitate for each sample. The dried starch was passed through a 250 µm sieve and stored in at −20 °C for further analysis.Starch (30.0 g) was weighed in a glass jar and 270 mL of distilled water was added to obtain a starch:water ratio of 1:9   was used as described by Alamri et al. , where sGel texture was determined according to Sandhu and Singh  using th×g for 15 min. The precipitate (W2) was weight and the swelling power was measured by the following formula: Swelling power = (W2 − W1)/weight of starch (g/g).The swelling power was measured according to the method described by Waliszewski et al.  with sliThe α-amylase activity in the GSE was estimated using the effect of pure α-amylase on the pasting properties of the tested starches. Therefore, 1.0% (5 mg/10 mL) α-amylase (EC3.2.1.1) was prepared and 0.1, 0.2, 0.3, 0.4, and 0.5 mL were added to 2.8 g of corn starch. Likewise, 0.1 mL of germinated sorghum extract (GSE) was added to 2.8 g of corn starch. Both corn starch samples were placed in a RVA aluminum can and the final weight (28 g) was completed with distilled water. The slurry was annealed for 30 min and the test was done in the RVA as mentioned above.p < 0.05). SAS Foundation 9.2 for Windows was used to analyze the data .All experiments were done in triplicate. Experimental data were analyzed using analysis of variance (ANOVA) and were expressed as mean value ± standard deviation. Duncan’s multiple range test was conducted to assess significant differences among experimental mean values (The amylose content (%) of wheat (W.S.), chickpea (C.P.), sweet potato (S.P.S.), Turkish beans (T.B.), and corn starch (C.S.) was 25.0% ± 0.07%, 24.0% ± 0.09%, 22.6% ± 0.06%, 20.9% ± 0.06%, and 20.4% ± 0.08%, respectively. Native starches exhibited peak viscosity as: 2828, 2438, 1943, 2250, and 4601 cP for the C.P., C.S., T.B., W.S., and S.P.S., respectively. Annealed starches without the addition of germinated sorghum extract (GSE) exhibited different pasting properties . It was During germination of sorghum, the activity of α-amylase increases evidently by the changes that occur during starch gelatinization, such as drop in the peak viscosity. After introducing GSE, the highest viscosity was recorded for the S.P.S. regardless of GSE concentration or annealing temperature, while the lowest was W.S. The viscosity of the remaining starches was in between S.P.S. and W.S. This indicates S.P.S. resistance to α-amylolysis under annealing conditions. At 60 °C, W.S. was completely gelatinized without GSE presence. In addition, C.S. and T.B. were the most heat sensitive when annealed at 40 and 50 °C, but after adding GSE, W.S. was the most sensitive and exhibited the least viscosity. What connects C.S. and T.B. is the similar amylose content despite their botanical origin. All starches, regardless of annealing time, exhibited lower viscosity at higher GSE. The enzyme attack on starch granules increases water absorption, which causes faster granule rupture, leading to lower viscosity. The opposite of this mechanism is when granules resist enzyme attack, which in turn prolongs rupture; consequently, granules maintain swelling for a longer time, and the viscosity subsequently increases. Based on this data, W.S. is the most susceptible among the tested starches due to the lowest viscosity, whereas S.P.S. was the least susceptible because of the highest viscosity followed by C.P. These two starches (S.P.S. and C.P.) did not have the highest amylose content among the tested starches. Variations in the degradation degree by α-amylase was observed among starch samples, indicating that subtle differences in the structure of starch granules with similar amylose content can influence α-amylolysis and the mechanism of hydrolysis.This occurrence is due to the ratio of amylose to amylopectin, as well as the degree of interaction between starch chains within the amorphous domains in the native granules as shown by XDR . In addip < 0.05) of W.S. to α-amylase compared to the other tested starches followed by C.P. Generally, starches with higher viscosity (less susceptible) exhibited higher breakdown without GSE, but by adding more GSE, the breakdown has increased due to the action of α-amylase that continued to degrade the starch fractions after the rupture of the granules.Therefore, granule structure and compactness of the amorphous region are the main cause for the difference between the data presented here and the literature ,26. CertIn the most part, annealed C.P. exhibited the highest setback 2553 cP), whereas C.S. (799 cP) and S.P.S (958 cP) exhibited the least setback; however, in the presence of GSE, W.S. had by far the least setback (568 cP) . These t53 cP, whThe final viscosity followed the same pattern as the setback where annealed W.S. ranked last (2896 cP) and C.P. first (5108 cP). Starch samples experienced drop in final viscosity at different rate at excess GSE, where C.P. and W.S. had the highest final viscosity drop the most in GSE, 95% and 78% for W.S. and C.P., respectively. The drop was sustained at the 60 min annealing as well, which indicates higher sensitivity to the enzyme degradation after gelatinization. The pasting temperature (PT) ranking of the native starch was: W.S. > S.P.S. > T.B. > C.S. > C.P., but after annealing without GSE, the ranking was W.S. > T.B. > C.S. > S.P.S. > C.P. Starch pasting after annealing at lower temperature indicates that annealing created porous granules, whereby quicker water uptake which leads to earlier pasting. This ranking did not match any of the other parameters except for W.S. swelling which supports the idea of the increased porosity after annealing. After annealing, the PT of most of tested starch samples stayed the same except for slight drop of W.S. from 84.1 to 72 °C, and slight increase for C.P. from 71 to 73 °C.Since the enzyme activity was not determined on the GSE, the following was done to estimate the amount of the enzyme in the GSE. RVA runs with pure α-amylase was compared to GSE. The pasting profiles of the samples with the GSE and the pure enzyme were overplayed on top of each other in order to match the GSE profile with the profile of the known fungal α-amylase enzyme concentration. The results indicated that by adding 0.1 mL of GSE, the profile matches the profile of the sample with 0.5 mL of the fungal α-amylase. Therefore, the concentration of the enzyme in the GSE was 5 mg/10 mL.p < 0.05) increased the swelling power of W.S. from the least to the most swollen (g/g) which can be attributed to increase in amylose packing in the amorphous region by closing the space between molecules and slowing the water uptake. Therefore, swelling power of S.P.S. is temperature dependent. The swelling power of annealed W.S. ranked first (6.86 g/g) with no GSE addition regardless of annealing time or temperature and S.P.S. ranked last with an average of 2.2 (g/g) g/g). The different behavior of the starches in GSE at 50 °C was also observed earlier during the discussion of the pasting properties, (p < 0.01) positively correlated with starch paste peak viscosity [The swelling power ranking of the native  starches was C.P. > S.P.S.> C.S. > T.B. > W.S., but after annealing at 40 °C, the ranking was W.S. > C.S. > C.P. > T.B. > S.P.S. It is obvious how annealing significantly  is the measure of the force needed to deform the gel during compression and is related to its structure and strength. Legume starches (C.P. and T.B.) exhibited the firmest gels among the five tested starches 677 and 481 (g), respectively, whereas after annealing, S.P.S. was the softest 80 (g), but W.S. gel was the softest in the presence of GSE 98 (g) , althougWheat starch was the most heat susceptible and liquefied after annealing at 60 °C without GSE. It was also the most swollen at lower annealing temperatures, and in the presence of GSE, the weakest gel was formed at higher temperatures. Sweet potato starch was the least susceptible to heat and GSE, the least swollen, and exhibited the highest peak viscosity. The native C.P. starch presented highest swelling power before annealing and after annealing at 60 °C for 60 min. All tested starches exhibited lower peak viscosity and softer gel in the presence of GSE, and the most change was observed for C.P. and W.S. at lower temperatures and S.P.S. at higher temperatures. The most setback was noted for the legume starches (C.P. and T.B.). Amylose content was not the determinant factor of α-amylase digestion of the annealed starches. The combined factors controlling enzyme susceptibility are granule structure and compactness. Although reports in the literature claimed that high amylose content correlated negatively with the swelling power of some starches, the data presented here indicated that high amylose (W.S.) correlate positively with high swelling power and negatively with peak viscosity. High swelling power of W.S. is consistent with faster enzymatic digestion rates and higher temperatures, which is controlled by rapid diffusion of enzymes into loosely packed granular structures even with the high amylose content. However, S.P.S. (lower amylose content) restricted water uptake due to the packed density of the amorphous region compared to W.S. (higher amylose content). Therefore, GSE is a straightforward low-cost starch hydrolyzer that can be used at industrial scale with further improvement of the germination process.


A new coronavirus infection, COVID-19, has recently emerged, and has caused a global pandemic along with an international public health emergency. Currently, no licensed vaccines are available for COVID-19. The identification of immunodominant epitopes for both B- and T-cells that induce protective responses in the host is crucial for effective vaccine design. Computational prediction of potential epitopes might significantly reduce the time required to screen peptide libraries as part of emergent vaccine design. In our present study, we used an extensive immunoinformatics-based approach to predict conserved immunodominant epitopes from the proteome of SARS-CoV-2. Regions from SARS-CoV-2 protein sequences were defined as immunodominant, based on the following three criteria regarding B- and T-cell epitopes: (i) they were both mapped, (ii) they predicted protective antigens, and (iii) they were completely identical to experimentally validated epitopes of SARS-CoV. Further, structural and molecular docking analyses were performed in order to understand the binding interactions of the identified immunodominant epitopes with human major histocompatibility complexes (MHC). Our study provides a set of potential immunodominant epitopes that could enable the generation of both antibody- and cell-mediated immunity. This could contribute to developing peptide vaccine-based adaptive immunotherapy against SARS-CoV-2 infections and prevent future pandemic outbreaks. A new coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has recently emerged as a human pathogen that causes fever, pulmonary disease, and pneumonia ,2,3. FolIn the postgenomic era, the availability of vast sequence data from pathogens and the advancement of computational prediction tools have greatly facilitated identifying potential immunogenic epitopes in pathogen proteins. This can be useful in designing vaccines against designated pathogens ,9. In reIn the present research, we used an immunoinformatics-based computational approach to mine the proteome of SARS-CoV-2 and subsequently identify immunodominant epitopes of SARS-CoV-2. Detecting immune responses that are based on specific immunodominant epitopes enables generating both antibody- and cell-mediated immunity against a specific pathogen. This can facilitate the fast and effective elimination of the pathogen. Using a variety of computational tools, we predicted all possible B- and T-cell epitope regions in SARS-CoV-2 protein sequences, and selected those regions where both B- and T-cell epitopes mapped. Notably, despite evidence that SARS-CoV-2 originated from bats, it phylogenetically diverged from SARS-CoV ,19. TherWe retrieved the whole genome and proteome of SARS-CoV-2 isolates from different geographic locations from Genbank (NCBI). Protein sequences of SARS-CoV and MERS-CoV were also collected from Genbank. The experimentally determined B- and T-cell epitopes of SARS-CoV were retrieved from the publicly available Immune Epitope Database (IEDB)  with theLinear B-cell epitopes are peptides with antigenic abilities that are bound by receptors on the surface of B lymphocytes and, thus, generate immune responses . We usedPredicting T-cell epitopes is important for identifying the smallest peptide in an antigen that is able to stimulate CD4 or CD8 T-cells to generate immunogenicity. Thus, the aim here is to identify peptides within antigens that are potentially immunogenic. MHC-peptide binding is considered to be the most important determinant of T-cell epitopes . MHC binIt is important to identify epitopes that are crucial for inducing protection and eliminate others in order to develop peptide-based vaccines. Protective antigens are able to induce an immune response. Thus, Vaxijen V2.0  was usedAn IEDB conservancy analysis tool was utilized in order to analyze the degree of conservation of SARS-CoV-2 B- and T-cell epitopes. The population coverage of T-cell epitopes was analyzed using tools available at the IEDB . The preThe allergenicity of immunodominant epitopes were predicted using AllerTOP v. 2.0  and AlgPPeptide epitopes of various lengths (ranging from 7 to 20 residues) which presented on MHC Class I and II molecules were retrieved from SCEptRe  , AutoPepBackbone conformations of the peptides that were bound to human leukocyte antigen (HLA) proteins were collected from PDB, clustered, and used as structural templates for 3D modeling of the epitopes (identified as immunodominant in the immunoinformatics study). Based on similarities and common structural patterns in HLA-peptide binary complexes, we generated 3D structures of the epitopes that are listed in Docking grids were generated by the autogrid module of AutoDock4 application , using tvs. T-cell epitopes and selected those epitopes with 100% sequence coverage. The lengths of the immunogenic regions were selected based on the maximum coverage of B-cell or T-cell epitopes in the mapped regions. Immunodominant epitopes, which can generate both antibody- and cell-mediated immunity, were identified to generate memory cells against SARS-CoV-2. We first predicted B- and T-cell epitopes and their possible MHC alleles from the SARS-CoV-2 protein using a variety of tools described in the Methods in order determine immunodominant epitopes . All of in vivo. Therefore, it is necessary to test the predicted epitopes in vivo to ensure that they can generate B-cell and/or T-cell responses. Detailed understanding of protective immune responses against SARS-CoV might be presumably important for developing a vaccine against SARS-CoV-2 [We predicted the abilities of the epitopes to serve as protective antigens using Vaxijen and to understand the immunomodulatory effect of epitopes identified from immunogenic regions . Unique RS-CoV-2 . For thi2, Interestingly, the mapping of immunogenic regions onto the structure of SARS-CoV-2 spike glycoprotein C revealeSelection pressure of the human immune system has been shown to drive viral point mutations that evade immune surveillance . TherefoHuman leukocyte antigens (HLAs) are the most polymorphic genes in humans, and their allele distributio and expression vary by ethnic group and geographical location. The classical HLA loci are class I  and class II  molecules, which provide antigen presentation to CD8 and CD4 T-cells . TherefoEpitope allergenicity is a prominent obstacle for vaccine development. We thus verified that the identified epitopes are not allergens. The allergenicity analysis results of the seven immunodominant epitopes  highlighEpitopes are faced with extremely complex and competitive environments that include the multitude of HLA proteins that bind immunogenic peptides with different affinities, and present selected epitopes to surface receptors on immune cells. Therefore, we performed molecular docking analysis to understand the binding interactions of the identified immunodominant epitopes with human MHC complexes. Structures of different HLA-peptide complexes from MHC class I and II were collected and aligned, as described in Methods. Structures of HLAs are fairly similar within each group (I and II) and share the same canonical fold. The epitopes were clustered in similar conformations in the HLA antigen binding grooves created by two helices in parallel orientation A,B. For Theoretically, six types of peptide-MHC structures were possible: (1) peptide-HLA (MHC I), (2) peptide-HLA (MHC II), (3) peptide-HLA-TCR (MHC I), (4) peptide-HLA-TCR (MHC II), (5) peptide-HLA-BCR (MHC I), and (6) peptide-HLA-BCR (MHC II). In this study, types 1, 2, and 3 were considered. We modeled the binding of the epitopes to different HLA proteins from MHC class I and II, and to HLA-TCR (MHC I). In the peptide-HLA-TCR type of binding, the docking scores were mostly higher (as compared to the binary peptide-HLA complexes). This was because epitope molecules were confined to the interface area between their cognate HLA/TCR proteins C. This mKTFPPTEPK bound to HLA-A*1101 (PDB ID 1x7q) as the reference state, we performed an extensive conformational sampling and docking study of this complex. We demonstrated that the top-score docking peptide conformations were clustered around the native conformation, with an estimated energy −9.97 kcal/mol (corresponding to the nanomolar affinity range). Moreover, we found similar binding energies  in docking simulations of KTFPPTEPK binding with HLA-A*02:01 (epitopes from KTFPPTEPK) was identical to region Lys362–Lys370 in the SARS-CoV nucleocapsid protein. In the SARS-CoV-2 variant, this motif was predicted in the epitope sequences LNKHIDAYKTFPPTEPK, KHIDAYKTFPPTEPKKDKKK, and YKTFPPTEPKKDKKKK, corresponding to positions Lys361 to Lys369  is less pronounced . Vice versa, extended peptides LNKHIDAYKTFPPTEPK (length 17), KHIDAYKTFPPTEPKKDKKK (length 20), and YKTFPPTEPKKDKKKK (length 16) do not fit HLA binding sites in HLAs from MHC Class I. Interestingly, we found that the core part (KTFPPTEPK) of the LNKHIDAYKTFPPTEPK peptide can bind to the recognition site of HLA proteins from MHC Class I , whereas the N-terminal part of this 17-residue peptide is arranged outside the A-pocket. The C-terminal part was found to occupy the F-pocket of the binding site  [Using the crystal structure of the nonapeptide ing site D. In agrI system . We predI system  for all K length , and YKTVaccination is an effective way to improve public health by building up adaptive immunity to a target pathogen . HoweverIn this present study, we exploited immunoinformatics-based approaches to identify potential immunodominant epitopes from SARS-CoV-2, which could be useful for developing vaccines for the COVID-19 disease. The vaccines should be capable of activating both humoral and cellular immune responses in humans. Our approach to defining immunodominant epitopes entails identification of overlapping regions of B-cell and T-cell epitopes (MHC-I and MHC-II) from proteins of SARS-CoV-2, particularly at those sites where these epitopes are 100% identical to the experimentally-validated epitopes of SARS-CoV. We identified 15 potential immunogenic regions from three proteins of SARS-CoV-2, and mapped 25 epitopes that are 100% identical to experimentally validated SARS-CoV epitopes. Among 25 potential immunodominant epitopes identified containing 9–28 amino acid residues, the lengths of most of the epitopes were 16–18 residues. To understand the binding patterns of epitopes with MHC-I and MHC-II, we performed structural and molecular docking analyses. We found that in the library of our immunogenic sequences, epitopes 9–11 residues in length were mostly recognized by HLA proteins from MHC Class I, whereas longer epitopes tended to bind to MHC Class II proteins with higher affinities. This finding is in agreement with known canonical preferences. Further analysis of MHC class I processing reveals that epitopes of longer sequences can undergo proteasomal processing and that the core part of the region for MHC class I recognition within the epitope can be presented on the cell surface for surveillance by CD8 T-cells. An analysis of the population coverage by HLAs revealed seven epitopes among the predicted 25 immunodominant epitopes that are found in more than 87% of the global infected population, and show high binding affinity to MHC-I and MHC-II, as evidenced from structural and docking analysis. Furthermore, these seven epitopes were predicted as being non-allergen, non-toxic, and of low risk of triggering autoimmune responses, which highlights their potential as successful vaccine targets. The viral epitopes that are least likely to mutate should be selected in order to develop an effective vaccine. Thus, we analyzed available SARS-CoV-2 genomes from various geographic locations to identify the percentage of mutations in suggested epitope regions. We found evidence of point mutations in a few epitopes of SARS-CoV-2 isolates from the USA. This suggests that human immune pressure-induced genetic drift plays a central role in the genetic adaptation of SARS-CoV-2. Interestingly, we did not find any point mutations in the mentioned seven potentially immunodominant epitopes. This result indicates that these seven epitopes are potentially effective vaccine candidates. Hence, the development of vaccines using these seven immunodominant epitopes could activate both humoral and cellular immune responses in humans, and that these epitopes could cover almost all of the worldwide population. Our results thus offer important insight for the development of a peptide vaccine for COVID-19. The COVID-19 outbreak is an emerging threat across the globe. Despite this, there are currently no permanent antiviral drugs or vaccine reported for fighting this disease. In the present study, we identified immunodominant epitopes from SARS-CoV-2 proteins that could induce both humoral and cell-mediated immune responses in humans, using the most comprehensive immunoinformatics approaches. Molecular docking of the immunodominant epitopes with HLA alleles supports their higher binding affinities within different HLA alleles. Further, seven potential immunodominant epitopes were shortlisted based on their higher conservancy, higher global population coverage, and significant interaction to MHC class I and class II alleles with high affinity. These epitopes have a low risk of being allergen, toxic, or generating autoimmune reactions. These finding highlight that these seven immunodominant epitopes could be the potential vaccine targets against SARS-CoV-2. The computational approaches that were used in this study could be a benchmark for the identification of immunodominant epitopes from other emerging pathogens, particularly, coronaviruses, in order to develop potential universal vaccines against various new strains.


A-type corn starch; B-type potato starch; and C-type tapioca starch). Structural properties of the starches were studied by X-ray diffraction analysis. Mechanical activation in a planetary ball mill reduces the degree of crystallinity in proportion to pretreatment duration. C-type tapioca starch was found to have the highest degree of crystallinity. Energy consumed to achieve complete amorphization of the starches having different types of crystalline structure was measured. The kinetic parameters of the process (the effective rate constants) were determined. The rate constant and the strongest decline in the crystallinity degree after mechanical activation change in the following series: C-type starch, A-type starch, and B-type starch.This paper examines the effect of mechanical activation on the amorphization of starch having different types of crystalline structure ( Starch is a high-molecular-weight plant-derived polysaccharide that consists of amylose and amylopectin and occurs in many crops, such as potato tubers, corn kernels, and cassava roots ,2. StarcStarch consists of two types of polysaccharides. Amylose is a linear polymer, while amylopectin is a branched one. The crystallinity depends on the degree of structural ordering in a solid body. The crystallinity of native starch granules typically is 14%–45%; that it why there are a number of challenges related to its use and processing  is the current content of the crystalline phase of starch; [S]0 is the initial content of the crystalline phase of starch; t is the duration of mechanical activation; and k is the effective rate constant of the process.To determine the kinetic parameters of starch amorphization, this process can be described using the kinetics of a pseudo-first-order reaction :(3)Scr→ C-type tapioca starch has the highest rate constant of amorphization, while those for A- and B-type starches are comparable and twice lower. The C-type crystalline structure is formed by coexisting A- and B-type crystallites. According to the concept regarding the nature of mechanical impact exerted on polymeric materials and composites, it is fair to assume that when mechanical impact is applied to an object with a heterogeneous structure (C-type starch), the resulting stress wave front propagates nonuniformly. Therefore, crystal structure disordering takes place at a faster rate than when processing an object having a homogeneous structure (A- and B-type starches). Mechanical pretreatment of homogeneous A- and B-type starches is less efficient because of the uniform propagation of the deformation wave. Therefore, it is fair to say that the A- and B-type starches are more resistant to mechanical impact. The crystallinity degree significantly affects the physicochemical properties such as hardness, density, transparency, and reactivity. However, these properties do not depend solely on crystallinity degree and are also affected by granule size and the type of crystal structure. Mechanical pretreatment results in particle refinement and increases the amount of the amorphous phase of starch, eventually improving the solubility and reactivity of the sample. These properties were employed when conducting enzymatic hydrolysis of mechanically pretreated biomass and developing novel types of functional foods characterized by improved nutrient bioavailability .B-type structure), while the highest one, for tapioca starch (C-type structure). The effective rate constants, parameters depending on the crystalline structure of starch, were calculated in order to describe the amorphization kinetics. Tapioca starch having the C-type structure is characterized by the strongest reduction of its crystallinity degree after mechanical activation and the highest amorphization rate constant. Hence, mechanical activation of A-, B-, and C-type starches in a planetary ball mill under various conditions alters their degree of crystallinity, thus broadening the range of their industrial applications. The increased reactivity of starch was utilized to develop novel types of functional foods characterized by better nutrient bioavailability.The effect of mechanical activation in a planetary ball mill on amorphization of starch depending on the type of its crystalline structure has been studied in this work. It has been shown that increasing duration of mechanical activation gives rise to samples with various degrees of crystallinity, as well as to the completely amorphized sample. Power consumption and energy consumed for mechanical activation have been measured. The energy consumed for amorphization of starch samples depends on the type of crystalline structure of starch. The lowest value was observed for potato starch (


Radiologia Brasileira (RB) and getting it indexed in international databases has long been a priority for its editors and for the Board of Directors of the Colégio Brasileiro de Radiologia e Diagnóstico por Imagem . Those have also been the objectives of practically all major national and international medical journals.Improving the journal .The Brazilian Coordenação de Aperfeiçoamento de Pessoal de Nível Superior  recently promoted changes to its graduate program evaluation system, bringing to the national scenario in Brazil what is already a reality in the international scientific publishing community: the growing role of Scopus/Scimago, a database created by Elsevier, which is becoming increasingly more well structured and comparable to the more traditional, more widely used, and until recently unique Web of Science database , from which the Journal Citation Reports (JCR) is produced on an annual basis. The JCR is the most commonly used citation reference for calculating the impact factor of a journal. The Scimago database has established itself by covering a larger number of scientific publications, including all of those indexed in the Web of Science database. In comparison with that of the Web of Science, the Scimago database covers approximately twice as many journals from three times as many countries,3.Scientific journals are primarily classified, in terms of quality, by their impact factor (as published in the JCR) or by its Scopus equivalent, the Scimago CiteScore, which is based on the number of citations per document (cites/doc). Both indicators are based on the use of a similar formula: the measure of the impact of a journal is determined by calculating the number of citations received by the articles it published in a given year or number of yearsRB in the Scopus/Scimago database (in 2012/2013) and in PubMed (in 2015), its Scimago CiteScore has been progressively increasing, from 0.669 in 2014 to 2.868 in 2018. That places RB in a privileged position in the Scimago ranking. If we consider the cites/doc for the last two years, it is currently ranked 59th among 328 radiology journals worldwide and is now the highest-impact journal among the 207 indexed medical journals of Latin America.Since the indexing of RB papers in our scientific articles published in other indexed journals. It should also be noted that only citations from the last two years are of interest-3. The growing prestige of the journal is evidenced by the fact the number of its citations in other indexed journals rose from 35 in 2014 to 192 in 2018, an increase of 450%.Authors and researchers in the field of radiology, especially those pursuing a master’s or doctoral degree in a graduate program evaluated by CAPES, benefit greatly from the advances in the qualification of our journal, not only in terms of the evaluation of the graduate programs in which they are enrolled but also in terms of their personal evaluations when they apply for research grants from funding agencies. However, everyone can contribute to advancing this process. The only way for the journal to continue to rise in this ranking and to increase its impact further is to raise our awareness regarding the importance of citing RB Editors and the CBR Board of Directors are counting on everyone to help the journal play a solid, consistent role on the international stage and, much more importantly, become increasingly more useful to and enjoyable for its readers.The


Translucent zirconia, manufactured by increasing the yttria content, offers improved translucency, but may have a negative effect on esthetic outcomes under clinical conditions such as discolored abutment because of the reflection of the underlying color. The purpose of this in vitro study was to compare the translucency parameter and masking ability of 3 mol % yttria-stabilized tetragonal zirconia polycrystal (3Y-TZP (Katana HT)), 4Y-ZP (Katana STML), and 5Y-ZP (Katana UTML) with those of lithium disilicate (Rosetta SM). Zirconia and lithium disilicate specimens of 10 mm diameters and 0.8 and 1.5 mm thicknesses were fabricated. Their CIE L*a*b* values  were measured at the center of the specimens against black and white backgrounds using a spectrophotometer, and translucency parameter (TP) values were determined. The microstructure of the specimens was observed using scanning electron microscopy. Four cylindrical backgrounds of different shades were prepared. The zirconia and lithium disilicate specimens were placed on the backgrounds without any intervening medium. CIE L*a*b* values were obtained, and the color difference value (ΔE) was calculated. Thresholds for acceptability and perceptibility were assumed as ΔE = 5.5 and ΔE = 2.6, respectively, to evaluate masking ability. Data were compared using one-way analysis of variance and post-hoc was performed using Scheffe’s test (α = 0.05). In zirconia specimens, the TP value increased as the yttria content increased from 3 mol %, through 4 mol % to 5 mol %, and all zirconia specimens showed lower TP values than lithium disilicate specimens did. All zirconia specimens showed optimal masking ability against a normal dentin shade (ND3) and acceptable masking ability against titanium at a minimum thickness of 1.5 mm. However, no zirconia specimen could mask severely discolored dentin (ND9), regardless of thickness. The decrease in zirconia thickness from 1.5 to 0.8 mm significantly increased translucency. Monolithic Y-TZP ceramics could mask a normal dentin background but could not mask severely discolored dentin at either 0.8 or 1.5 mm thicknesses. Zirconia is a bioinert and biocompatible material with high mechanical strength, good chemical stability, and low thermal conductivity ,2,3,4. FDespite its excellent mechanical properties, the opaque nature of 3Y-TZP is its significant disadvantage. This esthetic deficiency has conventionally been managed by the use of a layer of powder-fire porcelain on a zirconia core, but bilayered structures are vulnerable to chipping and delamination, exacerbated by thermally induced residual stresses . AttemptIn an attempt to develop a monolithic ceramic with acceptable translucency, 3Y-TZP processing was refined by reducing the concentration of alumina and minimizing the porosities by sintering at a higher temperature ,8. UsingMore recently, dental zirconia has been manufactured with increased yttria content to overcome the esthetic deficiencies. Partially stabilized zirconia with higher yttria content comprises an increased nonbirefringent cubic phase . Doping However, translucent materials may have a negative effect on esthetic outcomes under clinical conditions such as discolored tooth abutment, metal core, or titanium implant abutment, as the underlying color would be reflected. The effects of the background on the color of porcelain veneers ,12 and lTherefore, the purpose of this in vitro study was to compare the translucency parameter and masking ability of zirconia materials exhibiting differing yttria contents with those of lithium disilicate ceramics. First, we compared the translucency of 3Y-TZP (Katana HT), 4Y-ZP (Katana STML), and 5Y-ZP (Katama UTML) with that of lithium disilicate (Rosetta SM). Thereafter, we compared the masking ability of these zirconia materials with that of lithium disilicate on three different backgrounds.The zirconia materials selected for this study were Katana HT, STML, and UTML , exhibiting differing yttria content. Rosetta SM  was selected as the reference lithium disilicate material . The shaFour backgrounds of different shades were prepared to compare masking ability. Cylindrical substrates simulating different backgrounds included A2-shade composite resin  for reference, ND3-shade composite resin  for normal dentin, ND9-shade composite resin  for severely discolored dentin, and milled titanium for implant abutment . All bacTo obtain the value of the translucency parameter (TP), the CIE L*a*b* values  were measured at the center of the specimens on white and black backgrounds using a spectrophotometer . The values of each specimen were measured twice and averaged. The TP value was the color difference between values on the black and white backgrounds of the same specimen. The TP value was calculated using the following formula:The microstructures of the zirconia specimens were observed under a scanning electron microscope . For pretreatment, the specimens were degreased, dried, cleaned, and coated with platinum ions.The zirconia and lithium disilicate specimens were placed on four different backgrounds without any intervening medium . The CIEThe thresholds for acceptability and perceptibility were assumed as ΔE = 5.5 and ΔE = 2.6, respectively, to evaluate masking ability ,18,19.Statistical software  was used for statistical analysis of the data. The means of TP and ΔE values for each group were compared using one-way analysis of variance (ANOVA) and post-hoc was performed using Scheffe’s test. The level of statistical significance was set at 0.05.For the 0.8 and 1.5 mm thick zirconia specimens, the TP value increased significantly with the increase in yttria content from 3 mol % (HT), through 4 mol % (ST) to 5 mol % (UT). The TP values of zirconia specimens were less than those of lithium disilicate (RS) specimens .p < 0.001), which was significantly lower than the mean TP values in the UT group (15.36 ± 0.50) (p < 0.001) and the RS group (19.18 ± 0.29) (p < 0.001). There were statistically significant differences between all groups. The transparencies of specimens in the HT, ST, and UT groups were approximately 60%, 73%, and 80%, respectively, that of specimens in the RS group.For the 0.8 mm thick specimens, the mean TP value of specimens in the HT group (11.58 ± 0.57) was significantly lower than that of those in the ST group (13.90 ± 0.17) (p < 0.001), the ST group was significantly lower than the UT group (12.64 ± 0.19) (p < 0.001), and the UT group was lower than the RS group (14.20 ± 0.39) (p < 0.001). There were statistically significant differences between all groups. Similarly, the transparencies of specimens in the HT, ST, and UT groups were approximately 54%, 82%, and 89%, respectively, that of specimens in the RS group.As thickness increased, the TP value decreased, implying that opacity increased. In 1.5 mm thick specimens, the mean TP value of specimens in the HT group (7.75 ± 0.57) was significantly lower than that of those in the ST group (11.68 ± 0.23) (p < 0.001). In the zirconia groups, as yttria content increased, ΔE value increased; the RS group showed the highest ΔE value.Mean ΔE values of the specimens in the zirconia and lithium disilicate groups for the tested backgrounds are presented in Against the ND3 background, ΔE values of the specimens in the zirconia groups were under 2.6, regardless of thickness, implying optimal masking ability . The meaAgainst titanium backgrounds, all 1.5 mm thick specimens showed acceptable masking ability . HoweverAgainst the ND9 backgrounds, i.e., severely discolored dentin, only 1.5 mm thick specimens of the HT group (4.33 ± 0.45) showed acceptable masking ability .Translucency is defined as the relative amount of light transmission or diffuse reflection from a substrate surface through a turbid medium . The optThe translucency and opacity of materials depend on the extent of reflection, scattering, refraction, transmission, and absorption of incident light. Light transmitted to the interior of Y-TZP may exhibit interior reflection and refraction. This phenomenon is termed internal light scattering, which may result from several sources, including pores, impurities, defects, and grain boundaries.  In the cThe calculation of the translucency parameter is one of the most common approaches to assessing light interactions in dental materials. The translucency parameter has the advantage of direct visual evaluation of translucency . In thisConcerning translucency, thicker ceramic specimens exhibited lower TP values, a finding in line with those of previous studies ,26. In aHowever, the TP values of the specimens in the Y-TZP groups were lower than those of the specimens in the RS group; hence, monolithic anterior restorations would be limited to the highly translucent lithium disilicate ceramics.According to Yu et al., the TP value of human enamel at 1 mm thickness is 18.7, and that of dentin is 16.4 . In thisIn this study, ∆E values of the zirconia specimens on A2 and three other shades of abutments were obtained to evaluate shade-masking ability. The results showed significant differences in CIE L*a*b* and ΔE values, according to ceramic thickness and background type. Choi and Razzoog studied the masking ability of zirconia against four backgrounds and found a greater ΔE value with the black background than with the tooth-colored background . This reSeveral previous studies have found that not only abutment shade but also ceramic thickness affects the final shade of the restoration ,16,25. TConcerning masking ability on the ND3 background, 0.8 mm thick specimens of the HT, ST, and UT groups showed optimal masking ability; however, the mean ΔE value of the specimens in the RS group (3.36) was above 2.6, i.e., it did not show ideal masking ability. The transparencies of the 0.8 mm thick specimens in the UT group were approximately 80% of those in the RS group; hence, UT could be an acceptable alternative to RS. In the ST and UT groups, a thickness of 1.5 mm was required to mask the titanium background, and ΔE values were similar to those of the RS group. The transparencies of 1.5 mm thick specimens in the UT group and similar specimens in the ST group were almost 90% and 80%, respectively, that of specimens in the RS group. Considering both transparency and masking ability, UT and ST could be good alternatives for restorations against titanium abutments. Overall, the results of this study showed that 0.8 mm thick UT specimens showed masking ability and TP almost similar to those of RS specimens of similar thickness. Choosing the correct ceramic material with appropriate thickness might be the best method for masking background color, and the decision should be made based on mechanical and esthetic requirements.A limitation of this in vitro study was that it did not consider the effects of dental cements. Malkondu reported that regardless of cement type, color changes occur after cement application to monolithic zirconia . TranspaA decrease in zirconia thickness from 1.5 to 0.8 mm significantly increased translucency; however, all specimens showed translucency lower than that of human enamel.The TP value increased significantly with an increase in yttria content (mol %) of zirconia.5Y-ZP showed, approximately, 80% translucency at 0.8 mm thickness and 89% at 1.5 mm thickness, compared to that of lithium disilicate.Monolithic CAD-CAM ceramics could mask a normal dentin background but could not mask severely discolored dentin at either 0.8 or 1.5 mm thicknesses.Within the limitations of this in vitro study, the following conclusions could be drawn:


The above article published online on 02 October 2019 in Wiley Online Library (wileyonlinelibrary.com), has been retracted by agreement between the authors, the journal Editor in Chief Jun Chen, and John Wiley & Sons Ltd. The retraction has been agreed due to unreliable data and consequently its misleading results and conclusions.Retraction: Receptor tyrosine kinase AXL is correlated with poor prognosis and induces temozolomide resistance in glioblastoma, CNS Neuroscience & Therapeutics 2019, (


This article concerns the elaboration and the characterization of a novel biobased potato starch-beet pulp composite for packaging applications as cushion foams. A twin-screw extruder was used to elaborate composite foams. SEM observations of these materials were conducted, and thermomechanical properties were studied in terms of thermal transitions  and viscoelastic properties (DMA). The effect of relative humidity content on viscoelastic properties was analyzed as a function of frequency. The different test results show that the composite structures are homogeneously mixed. The sponge-like structure of the beet-pulp disappears indicating a good compatibility between the two mixed constituents. The DSC curve of starch-beet pulp foam shows a single thermal transition at 153.6 °C, indicating the thermal homogeneity of the obtained composite material. The density value of starch-beet pulp foam is higher than conventional foams, but this can be optimized by adjusting the technological parameters of the extruder. The viscoelastic properties of the developed materials depend on the relative humidity. The packaging industry is looking for materials with specific properties  that can be easily disposed of after use while being environmentally friendly. At the same time, consumers have developed an environmental conscience and have now become aware of the need to use biodegradable packaging . PackagiThe main objective of this exploratory study is the elaboration and the characterization of a novel foamed biobased composite made of beet pulp and potato starch used as cushioning for industrial goods . In thisIn this study, starch-beet pulp composite foams were prepared by extrusion. The composites were characterized for physical, structural, and mechanical properties.The extruded beet pulp (BP) pellets were purchased from Cristal Union factory  with 8–10 mm diameter and 18% humidity  during tPotato starch (15% humidity) was provided by Roquette . This starch was chosen among other botanical sources because of its high polymerization degree and high viscous properties providing interesting mechanical properties for the Starch-Beet Pulp (SBP) composite. In SBP composite starch acts as a binder to stick the particles together and assure the transfer of shear forces between the fibers.3, provided by A.D society (France), was used as a plasticizer. The obtained admixture was mechanically stirred to homogenize the distribution of the different constituents in the SBP blend.Potato starch and potato SBP composite foams were prepared as follows: to 10 parts of dried swollen pulp obtained from extruded beet pulp was added 3 parts of water and 10 parts of a 7/5 starch-glycerol mix previously dried for 2 h at 120 °C in an oven. Glycerol with a density of 1.26 g/cmThe SBP mix were compounded using a Clextral BC21 co-rotating twin-screw extruder machine. The configuration of a mixing and suited transport screw elements is shown in −3 g. The absolute density is calculated using the following equation:−3), −3), Bulk density, also called apparent density or volumetric density, is defined as the mass of the material divided by the total volume it occupies. It was calculated as the mass of an extruded SBP (ESBP) ribbon divided by its geometric external measured volume. Absolute density is obtained when the volume measured excludes the pores as well as the void spaces between material within the bulk sample. In this study, the absolute density was determined using the Bourdot et al. pycnometer method  particulScanning electron microscopy (SEM) observations were carried out on a 15kV JEOL/JSM 6460LA (JEOL (Europe) SAS, Croissy, France). All samples were dried at 60 °C for 48 h in a vacuum desiccator, then coated with carbon to increase their electrical conductivity.TGA experiments were conducted using a TGA (NETZSCH-TG 209 F3 Tarsus)  to evaluate the moisture content and the thermal stability of potato starch and starch-beet pulp composite foams. The measurements were carried out in triplicate on 10–15 mg samples put in an aluminum pan and heated from 25 °C to 600 °C with a heating rate of 10 °C/min in a nitrogen atmosphere.The thermal characteristics of potato starch and starch-beet pulp composite foams were determined by using a DSC (NETZSCH – DSC 204 F1 Phoenix) , after 48 h conditioning at a temperature of 23 °C and 65% relative humidity. Aluminum sealed pans were used to test in triplicate samples of 5–40 mg. The heat flow was measured as function of the temperature and time. The samples were heated from −80 °C to 200 °C at a heating rate of 20 °C/min. A reference empty DSC pan was used to balance the heat capacity of the sample pan. Proteus V4.8.5 NETZSCH software is used to analyze the results.DMA tests were carried out in a DMA Q800 , with DMA-RH accessory for humidity and temperature control. This makes it possible to determine the mechanical properties of the samples under controlled humidity and temperature conditions.2 circular slices of 5 mm thickness. The samples were preloaded with a force of 0.01 N and then assigned a constant stress of 10 kPa for 5 min at a constant temperature of 23 ± 2 °C and variable relative humidity conditions ranging from 30% to 52% RH. After 5 min, the stress was removed, and the samples undergo 15 min recovery. The creep compliance of the tested materials was measured.To compare the viscoelastic properties of the different foams, creep/recovery tests were performed using the compression clamp kit which is suitable for low to moderate modulus materials such as foams. In this mode, the sample is placed on a fixed flat surface of 40 mm diameter and an oscillating plate applies a defined force. The samples were cut into 1.22 cmTo assess the viscoelastic properties as a function of frequency, we have also performed tests in the multi-frequency mode (0.1 Hz–100 Hz), with a constant oscillation stress amplitude of 10 kPa at a constant temperature of 23 ± 2 °C and variable relative humidity conditions ranging from 13% to 54% RH. The storage and loss moduli of the materials were measured.The following SEM images make it possible to visualize the microscopic structure of the raw materials and their physical transformation after the extrusion process.The dry lyophilized starch raw material SEM micrographs are shown in After plasticization of starch, the cell structure observed in Beet pulp has a homogeneous sponge-like native structure as shown by the SEM micrographs given in The potato starch-beet pulp composite foam structure is given in 2 for example) can also be added in the extrusion process to improve the foam expansion and decrease the density value.TGA thermograms of beet pulps are given in DSC is one of the most used techniques for the analysis of thermal transition of polymers and biopolymers such as starch-based and beet pulp-based materials ,31. TypiThe storage modulus represents the energy stored in the elastic structure of the sample. When it is higher than the loss modulus the material can be considered as mainly elastic. The loss modulus is the viscous part and represents the amount of energy dissipated in the sample. The storage (G’) and loss (G”) moduli of starch-beet pulp foams at different RH conditions are given in The creep-recovery curves of starch-beet pulp composite foams at different RH conditions are plotted in The creep-recovery response of the foams can be modelled using the viscoelastic generalized Kelvin-Voigt model given by the following equations for creep (The obtained results are summarized in 2 for example) can also be added in the extrusion process to improve the foam expansion and decrease the value of the density.We proposed in this study a composite foam material extruded from beet-pulp and starch as an alternative to polymer foams such as expanded polystyrene (EPS), loose-fill (foamed chips for filling space around goods within a packing box) used in cushion packaging. The extrusion process of the beet-pulp/plasticized starch compound produces a homogeneous foamy material. These results were obtained with a non–optimized process and can be improved by adjusting the technological parameters at the exit of the extrusion die since the mixture tended to collapse after extrusion. A physical or a chemical agent such as an expanding gas (CO


Furthermore, Fourier transform infrared spectroscopy analysis demonstrated the interaction between starch and CTS, which confirmed that the compatibility between CTS and PS is excellent.The effects of chitosan (CTS) as the reinforcing phase on the properties of potato starch (PS)-based foams were studied in this work. The formic acid solutions of CTS and PS were uniformly mixed in a particular ratio by blending and then placed in a mold made of polytetrafluoroethylene for microwave treatment to form starch foam. The results showed that the molecular weight and concentration of CTS could effectively improve the density and compressive properties of starch-based foams. Furthermore, orthogonal experiments were designed, and the results showed that when the molecular weight of CTS in foams is 4.4 × 10 Conventional plastics are nondegradable, which degrade slowly and require a long time, even hundreds of years, to degrade completely. They also impact the soil negatively during this time. Therefore, the exploration of degradable plastics that can replace traditional plastics holds great practical significance.Global pollution is increasingly becoming a serious concern, causing extreme climate changes and natural disasters. The huge pollution of traditional plastic products in the environment has extensively raised people’s concern. Traditional plastics such as polystyrene (PS), soft (hard) polyurethane, polyvinyl chloride, and polyolefin are widely used as the matrix of foaming materials, but due to the poor operability and degradability of these plastics, most of the foaming materials are wasted, leading to increasingly serious white pollution. With people’s increased awareness of environmental protection, considerable attention has been paid to the development of environmentally friendly materials. Therefore, bio-based and biodegradable polymers as replacement materials for traditional nonbiodegradable plastics have been extensively studied ,2,3,4. B2O and CO2. Starch-based plastics are beneficial in being nontoxic, harmless, widely available, and cheap and also possess relatively superior properties. Use of these plastics effectively can alleviate the problem of white pollution and the crisis of biochemical energy shortage  is defined as the reduced specific viscosity (sp/cη) or the relative viscosity of the inherent relative viscosity (r/cln η) of a solution in the case of infinite dilution of solution mass concentration (c), which is expressed as follows: The intrinsic viscosity [η] of polymer solution exhibits a specific relation with the molecular weight of polymer at a specific temperature. According to the Mark–Houwink–Sakurada (MHS) equation, the following empirical formula is used for calculating [η] and the molecular weight of polymer solution ; the molecular mass of polymer can be calculated using Equation (3).The relationship between (2) Morphology of starch-based foamsThe starch-based composite structure was investigated using scanning electron microscopy (SEM)  . Foam samples were sputter-coated with platinum for 20 min after cutting with a razorblade into 3-mm thick slices, and the transversal cross-section area was examined using SEM.(3) Pore size, porosity, and density of CTS–PS foams3 size, and then the exact size of the sample was measured to ensure that the error was less than 0.02 mm. The sample mass was weighed with an electronic balance to make the result accurate to 0.001 g, and finally the density of the material was calculated using the following formula:ρ = density (g/cm3), M = foam weight (g), V = foam volume (cm3)Image analysis (Image-J) software  was used to measure the average diameters of nearly 100 pores. Plot of pore size distribution enabled the comparison of cellular structure for different formulations of starch foams. The finished foam was cut into five samples of 24 × 24 × 32 mm(4) Mechanical propertyAccording to the standard GB/T 8813-2008, the compressive strength of foam materials was tested using a CMT 4000 universal testing machine . The vertical direction of foam growth was consistent with the direction of compression. The compressive strength of the foam material is the ratio of the maximum compressive stress to the original cross-sectional area of the sample, when the relative deformation of the material is less than 10%, which is expressed in kPa.P = compressive strength (kPa), F = compressive stress (kN), S = cross-sectional area (cm3), The specific formula is as follows:(5) Fourier transform infrared analysis−1 was acquired for every sample. The processing software OMNIC 8.0  was used to perform automatic baseline correction of the collected infrared spectrum to ensure its baseline level.The dried samples and potassium bromide were ground into a fine powder in an agate mortar and pressed into tablets. The Fourier transform infrared (FTIR) spectrum of material was measured using the VERTEX 80 V type FTIR spectrometer  and the spectrum at a range of 4000–400 cm(6) Thermal characterizationThe total mass loss on a TG209F3 analyzer  was assessed using the thermogravimetric (TG) method. For the 6 mg samples dried for 24 h at 70 °C, the experimental temperature was set as 30 ± 3 °C to 800 °C in nitrogen atmosphere. The heating rate was 10 °C/min, and the experimental flow rate was 30 mL/min.(7) Differential scanning calorimetry (DSC) testThe glass transition temperature of sample was evaluated by a 200 F3 Differential Scanning Calorimeter . The dried sample of around 3 mg was placed in an aluminum crucible and sealed. An empty aluminum box was used as a control and nitrogen was used as purge gas. The temperature ranged from 20 to 180 °C and the heating rate was 10 °C/min.(8) Solubility of foams in waterThe samples were dried in a drying oven at 60 °C for 24 h and then soaked in deionized water at room temperature. The shape of the sample in water was recorded until it dissolved.η] was 502.39, and the molecular weight of the CTS sample calculated according to the MHS equation was 3.5 × 105. Likewise, the molecular weights of other two types of CTS were also estimated through the same calculation; 2) of CTS with different viscosities was greater than 0.95, which indicates the reliability of the calculated results. The molecular weight of CTS with viscosity between 200 and 400 mPa·s was 4.4 × 105 and that of CTS with a viscosity of 400 mPa·s was 5.2 × 105  (5 (CTS-3.5/PS) was relatively smaller (5 (-4.4/PS) e; the qu smaller a than thaller  is the main structure of esterified phosphate in PS, and its acidity is stronger than that of orthophosphate [3+ species increases beyond a critical value, synthesis of uniform CTS/PS foams becomes challenging and results in weak interaction at the boundary and poor compressive performance.The foams composed of CTS and starch exhibit such strength mainly because of the formation of a high intermolecular hydrogen bond between NHhosphate ,31. Therhosphate ; however2B2C3, implying that the CTS molecular weight of 4.4 × 105, CTS mass fraction of 4%, and the CTS/PS mass ratio of 3/4.2 are optimal for foam synthesis. We verified the compression performance of the CTS starch-based foam under these conditions, which was higher (1.077 mPa) than those observed in other conditions.To determine the effect of each factor on the foam compression performance, we further designed an orthogonal experiment and variance analysis. −1 for the dried PS corresponding to the characteristic absorption peak of the hydroxyl group in the starch. A moderate intensity peak at 2930 cm−1 was associated with –CH2 antisymmetric stretching vibration, and an absorption band at 1647 cm−1 was attributed to water absorption in the amorphous region of PS; the absorption peak near 1380 cm−1 was ascribed to –CH3 bending vibration and –CH2 twisted vibration. C–C, C–O, and C–H stretching vibrations and bending vibration of C–OH formed obvious absorption peaks in the 1300–800 cm−1 region of the spectrum. CTS molecules also displayed a strong and wide absorption peak at 3428 cm−1, corresponding to the stretching vibration of O–H and N–H; the absorption peaks at 1660, 1500, and 1425 cm−1 corresponded to amide-Ι belt stretching vibration, –NH2 deformation vibration, and –CH2 and –CH3 bending vibrations, respectively. The characteristic absorption peak at 1157 cm−1 has been ascribed to the CTS C–O–C asymmetric vibration [To study the interaction between CTS and PS, an infrared spectrum of each material was obtained. ibration ,34.−1 and the characteristic peaks of starch at 1646 cm−1 moved to a low-frequency region, and the intensity of the absorption band increased. Correspondingly, the absorption band strength of foam composites at 3413 cm−1 increased, which indicates that the hydrogen bond density in the CTS-starch system increased and inter- and intra- molecular hydrogen bonds formed between amino and hydroxyl groups in molecular chains of CTS and starch, thus improving the compatibility between CTS and starch [In a study, the interaction between CTS and starch in starch-based biological foam was shown to cause clear changes in the surroundings of polymer groups, which led to variations in the frequency and intensity of relevant absorption bands, and a unique response was obtained with changes in the characteristic spectra peak wave numbers . Relativd starch ,37.The thermogravimetric curves and corresponding thermogravimetric rate curves of raw materials and starch-based foams are shown in The second thermogravimetric stage (between 200 to 400 °C) involved the depolymerization of chitosan molecular chains and the cleavage of starch molecular chains; this included the dehydration of sugar rings and the depolymerization and decomposition of glucose units . Due to The DSC diagrams of chitosan and starch and CTS/PS foams are shown in Biological foams containing CTS and PS were prepared successfully in a new polytef mold in a microwave oven. The effects of different viscosities and concentrations of CTS and different mass ratios of CTS-starch on the morphological, physicochemical, mechanical, and thermal properties of CTS–PS foams were comprehensively studied. The research conclusions can be summarized as follows:5, 4.4 × 105, and 5.2 × 105. The results confirmed that high viscosity corresponds to high relative molecular weight of CTS.(1) The viscosity of CTS solution at a particular concentration directly reflects the molecular weight of CTS. The molecular weights of CTS with different viscosities were 3.5 × 105 exhibit small pores and low density. Moreover, the foams with a larger proportion of starch demonstrated small pore size and low density. Interestingly, the reverse was found to be true in the case of compressive properties; foams comprising CTS of molecular weight 4.4 × 105 exhibited the highest compressive strength, and foams with a larger proportion of starch exhibited higher compressive strength. In addition, orthogonal experiments were set up to confirm that the compressive performance of foams is highest when the molecular weight of CTS is 4.4 × 105, the mass fraction is 4 wt%, and the mass ratio of CTS-PS is 3/4.2.(2) Analysis of morphology, pore size, and density of starch-based biological foams indicated that CTS foams comprising CTS of molecular weight 4.4 × 10(3) The change in the characteristic peaks of the foams composed of CTS and PS in the FTIR diagram indicated that the amino group in CTS interacted with the hydroxyl group in the starch. In TGA and DSC analysis, it could be seen that the interaction between starch and chitosan improved the thermal stability of the foam, and in the water solubility test, the interaction maintained the integrity of the foam’s morphological structure in water within a certain period of time, and the foams completely degraded after 30 days.


Glioblastoma (GBM) is identified as a lethal malignant tumor derived from the nervous system. Despite the standard clinical strategy including maximum surgical resection, temozolomide (TMZ) chemotherapy, and radiotherapy, the median survival of GBM patients remains <15 months. Accumulating evidence indicates that rapid‐acquired radioresistance is one of the most common reasons for GBM recurrence. Therefore, developing novel therapeutic targets for radioresistant GBM could yield long‐term cures.To investigate the functional role of CXCL1 in the acquired radioresistance and identify the molecular pathway correlated to CXCL1.In this study, we identified that CXCL1 is highly expressed in GBM and the elevation of CXCL1 is involved in radioresistance and poor prognosis in GBM patients. Additionally, silencing CXCL1 attenuated the proliferation and radioresistance of GBM cells. Furthermore, we demonstrated that CXCL1‐overexpression induced radioresistance through mesenchymal transition of GBM via the activation of nuclear factor‐kappa B (NF‐κB) signaling.CXCL1 was highly enriched in GBM and positively correlated with poor prognosis in GBM patients. Additionally, elevated CXCL1 induced radioresistance in GBM through regulation of NF‐κB signaling by promoting mesenchymal transition in GBM. Additionally, knockdown of CXCL1 attenuated the proliferation and enhanced radiosensitivity of GBM cells. Furthermore, we demonstrated that CXCL1 induced mesenchymal transition of GBM via activation of NF‐κB signaling.22.12FC (Fold change) and adjusted P value, in which log2FC > 1 with an adjusted P value < .05 was identified as an upregulated gene. Venn diagram was carried out to find the overlapping upregulated genes in different datasets.Expression data of GSE56937 and GSE67089 were extracted from GEO database . Ninety‐one glioma samples and three nontumor tissue samples (from epilepsy surgery) were collected from patients underwent surgical operations from 2008 to 2016. All the patients' samples have obtained necessary consent. The samples were embedded in paraffin blocks as previously described.2.5Immunohistochemistry was performed as previously described.2.6shCXCL1#1: CCGGCAAATGGCCAATGAGATCATTCTCGAGAATGATCTCATTGGCCATTTGTTTTTGshCXCL1#2: CCGGGTTCTCCAGTCATTATGTTAACTCGAGTTAACATAATGACTGGAGAACTTTTTGLentivirus production and transduction was conducted as previously described.The CXCL1‐overexpression lentivirus was designed by Genechem.2.72 at 37°C. As for cell viability assay, cell number was calculated by cell counter with trypan blue, then cells were seeded into 96‐well plates after adequate suspension at a density of 2 × 103 cells/100 uL per well and cultured for 12 hours. After 24 hours of culturing at 37°C with 5% CO2, U87 and U251 cell lines received in vitro radiotherapy using X‐RAD 320 from Precision X‐ray at a dose of 8 Gy. Afterward, these cells were used to conduct indicated experiments. Finally, cell number was counted by alamarBlue and IC50 was calculated by using SPSS 22.0.Glioblastoma cell lines U251, SHG‐44, and NHA were provided by the First Affiliated Hospital of Xi'an Jiaotong University . U87 cell line was bought form BeNa Culture Collection. Cells were cultured in DMEM‐F12 containing 10% vol FBS and antibiotics (1% penicillin and streptomycin). All these cells were cultured in a humidified condition containing 5% CO2.85 cells in 2 uL PBS then injected into the nude mice brains as described previously.The usage of experimental animals in this study was approved by the Ethics Committee of the School of Medicine, Xi'an Jiaotong University . In vivo xenograft model was performed by using 6‐week‐old female nude mice. Corresponding GBM cell  was suspended and diluted to the density of 1 × 102.9® 488 Annexin V/Dead Cell Apoptosis kit strictly following the manufacturer's protocols.Flow cytometry assays were conducted as previously described.2.10high and CXCL1low (by quartile cutoff). Subsequently, limma packagehttps://david.ncifcrf.gov/).Gene expression profiles were derived from the Cancer Genome Atlas (TCGA) database. All data were preprocessed by background correction, gene ID transformation, and normalization by R. Furthermore, these data were ordered by the expression of CXCL1 to divide all samples into two groups, CXCL1high groups.As for gene set enrichment analysis (GSEA), the GSEA software2.11t tests were used to evaluate statistical differences. One‐way ANOVA was utilized in comparisons more than two groups, following Dunnett's post‐test. Kaplan‐Meier survival analysis was conducted using log‐rank analysis. All statistical analysis was calculated by GraphPad Prism 7 or SPSS 22.0 software. Unless specifically indicated, statistical significance was considered as a two‐sided P value < .05.All the results in this study are exhibited as mean ± SD (Standard Deviation). Number of independent replications is presented in the corresponding Figure legends. Two‐tailed 33.1P = .016, with log‐rank test) and ESM1 were significantly correlated to overall survival in GBM patients  were introduced into U87 and U251 cells. qRT‐PCR analysis illustrated that the CXCL1 mRNA expression was reduced more than 50% in both U87 and U251 transfected with shCXCL1 lentivirus compared with shNT cells Figure A and B. 3.4+; PI‐) and late  apoptosis were significantly diminished by CXCL1 overexpression followed by radiation, compared with radiotherapy alone  while 51 downregulated in CXCL1low group  was utilized to conduct the Gene Ontology (GO) annotation. Negative regulation of apoptotic process, positive regulation of cell proliferation, NF‐κB signaling, EMT, and ERK1/ERK2 cascade were the most enriched cancer‐related GO terms (Figure As we showed that CXCL1 functions as an oncogene and induces radioresistance in GBM, bioinformatics analysis of TCGA database was performed to further evaluate the underlying signaling pathways for CXCL1. Patient samples from TCGA database were grouped into two groups according to the expression of CXCL1. Hierarchical bi‐clustering analysis indicated significant gene signatures in CXCL1M Figure A. Throug4Glioblastoma is one of the most lethal tumors in human central nervous system. Despite the development of clinical strategies during the past several decades, outcomes of GBM patient is still remains dismal.DNA damage repair is regarded as a crucial process for GBM cells to acquire radioresistance.Recent studies have clarified that GBMs are highly plastic and have a natural tendency to transition from one subtype to another.In this study, we analyzed the transcriptome expression profiles of two published GEO databases related to radioresistance and molecular subtype classification of GBM. As a result, CXCL1 was identified as one of the most upregulated genes in both radioresistant cells as well as mesenchymal cells. Next, elevated CXCL1 expression was closely related to poor prognosis in glioma/GBM patients. Furthermore, molecular biological assays were conducted to demonstrate the potential role of CXCL1 by lentiviral silencing and overexpression, and we illustrated that CXCL1 promotes radioresistance of GBM while suppressing CXCL1 attenuated the migratory and invasive behaviors of GBM cells. Moreover, in vivo tumor xenograft mice model demonstrated that CXCL1‐knockdown prolonged the overall survival of tumor‐xenografted mice and increased the sensitivity of GBM. Additionally, EMT and NF‐κB signaling were proved to be the most correlated pathways of CXCL1 expression according to the results of GSEA by using TCGA GBM database, which was also confirmed by molecular biological assays. Overall, our findings suggest that the elevation of CXCL1 indicates a poor prognosis and radioresistance by inducing mesenchymal transition in glioblastoma.Although the potential role of CXCL1 in GBM radioresistance is well discussed in this study, additional researches on the molecular mechanism are still required for evaluating the clinical significance of CXCL1 inhibition in radiotherapy. Moreover, whether CXCL1‐induced radioresistance solely depends on its regulation in NF‐κB signaling and mesenchymal transition remains unclear. As GBM is a type of heterogeneous disease, inhibition of a single candidate biomarker might have unpredictable consequences. Therefore, in future studies, it will be critical to improve the overall management of GBM patients by focusing on specific subgroups of patients.5In conclusion, our findings suggest that CXCL1 is overexpressed in GBM and confers aggressive radioresistance to GBM cells, and suggest CXCL1 as a valuable biomarker for GBM patients. Both bioinformatic and functional analysis confirmed the role of CXCL1 in promoting radioresistance in GBM. We further identified EMT and NF‐κB as the downstream signaling pathways of CXCL1 and silencing CXCL1 attenuated tumor progression.The authors declare no conflict of interest. Click here for additional data file.