MPK38 phosphorylated p21 at Thr55, stimulating its nuclear translocation, which resulted in greater association of p21 with peroxisome proliferator-activated receptor γ (PPARγ), preventing the PPARγ transactivation required for adipogenesis. Furthermore, restoration of p21 expression by adenoviral delivery in diet-induced obese mice ameliorated obesity-induced metabolic abnormalities in a MPK38 phosphorylation-dependent manner. These results suggest that MPK38 functions as a positive regulator of p21, regulating apoptosis, cell cycle arrest, and metabolism during obesity.Murine protein serine-threonine kinase 38 (MPK38)/maternal embryonic leucine zipper kinase (MELK), an AMP-activated protein kinase (AMPK)-related kinase, has previously been shown to interact with p53 and to stimulate downstream signaling. p21, a downstream target of p53, is also known to be involved in adipocyte and obesity metabolism. However, little is known about the mechanism by which p21 mediates obesity-associated metabolic adaptation. Here, we identify MPK38 as an interacting partner of p21. p21 and MPK38 interacted through the cyclin-dependent kinase (CDK) binding region of p21 and the C-terminal domain of MPK38. MPK38 potentiated p21-mediated apoptosis and cell cycle arrest in a kinase-dependent manner by inhibiting assembly of CDK2-cyclin E and CDK4-cyclin D complexes  An understanding of its multi-site protein phosphorylation, a common mechanism for regulating protein function and stability, should provide insight into the mechanisms by which MPK38 regulates the activity of its substrates3. For example, MPK38 has been shown to phosphorylate Bcl-G, a pro-apoptotic protein, leading to the inhibition of Bcl-G-induced apoptosis4. MPK38 phosphorylates ZPR9, a zinc finger protein, and stimulates its nuclear localization, which results in greater B-myb transactivation5. Conversely, MPK38 phosphorylates PDK1 at Thr354, thereby inhibiting its activity and function6. Furthermore, MPK38 phosphorylates ASK1 (Thr838), p53 (Ser15), and Smads , activating the downstream pathways8. Phosphorylation of serine-threonine kinase receptor-associated protein (STRAP) at Ser188 by MPK38 activates ASK1/TGF‐β/p53 signaling and inactivates PI3K/PDK1 signaling, leading to apoptotic cell death9. However, Thr76 phosphorylation of thioredoxin (Trx) by MPK38 inhibits activation of ASK1/TGF‐β/p53 signaling10. Further study of such mechanisms should greatly enhance our understanding of the biological roles of MPK38.Murine protein serine-threonine kinase 38 (MPK38)/maternal embryonic leucine zipper kinase (MELK), a member of the AMP-activated protein kinase (AMPK)-related kinase family, has been proposed to be involved in the control of a variety of biological processes, including the cell cycle, cell proliferation, apoptosis, signal transduction pathways, tumorigenesis, and metabolism12. Consistent with its multiple cellular functions, a number of proteins have been found to interact with p21 in cells. In addition to the binding of p21 to cyclin-CDKs and proliferating cell nuclear antigen (PCNA)15, which leads to the inhibition of cell cycle progression and DNA synthesis, a variety of other proteins have been found to bind to p21; for example, transcription factors19, pro-apoptotic proteins20, protein kinases23, and many others28. p21 has also been shown to function as a positive regulator of the cell cycle; for example, p21 overexpression improves cell survival in the face of prostaglandin A2 or p53 overproduction30. p21 expression is also high in various tumors, particularly in late-stage, aggressive tumors, such as glioblastoma multiforme (GBM)33. In addition, p21 has been shown to play an important role in adipocyte differentiation: p21 knockdown by RNA interference in 3T3-L1 cells or its ablation in p21−/− mouse embryonic fibroblasts (MEFs) inhibits adipocyte differentiation, suggesting that p21 is a pro-adipogenic factor34. By contrast, other studies have shown that mice lacking p21 become obese as a result of adipocyte hyperplasia35, which implies that p21 may function as an anti-adipogenic factor. Thus, the precise role of p21 in adipogenesis remains controversial.p21, a cyclin-dependent kinase (CDK) inhibitor, was initially isolated as a p53-inducible protein and is known to regulate the cell cycle and DNA replication. It has also been implicated in various other biological processes, including apoptosis, cell differentiation, development, tumorigenesis, and metabolismIn this study, we show that MPK38 phosphorylates p21 at Thr55 and functions as a positive regulator of multiple p21 effects by stabilizing the protein. Moreover, we show that phosphorylated p21 significantly inhibits adipocyte differentiation and ameliorates obesity-induced metabolic abnormalities, implying that MPK38-induced Thr55 phosphorylation of p21 plays a role in the regulation of metabolism in obesity.7 and regulates the cell cycle36, we reasoned that MPK38 might regulate the activity of p21, a p53 target. To investigate this possibility, we first determined whether p21 physically associates with MPK38 using co-transfection experiments. p21 was only detected in the presence of glutathione S-transferase (GST)-tagged MPK38 but not control GST alone  using GST-tagged recombinant MPK38  and in vitro-translated act Fig. . In addiact Fig. , suggest32P]ATP and the phosphorylation of recombinant p21 protein was measured. p21 phosphorylation was identified in the presence of MPK38  of p21. The T55A mutation, but not the S116A and S153A mutations, completely abolished MPK38-mediated phosphorylation  MPK38  and adiponectin (Adipoq), was also lower in 3T3-L1 cells expressing WT p21 than in control cells expressing an empty vector alone , a PPARγ agonist. We found that WT p21 suppressed the transcriptional activity of PPARγ in a dose-dependent manner, whereas the T55A mutant, which is unable to translocate to the nucleus  reduced the number of large hypertrophic adipocytes  concentration, Ad-p21 infection reduced the mRNA expression of lipogenic genes in liver and WAT Fig. . Ad-p21 ins Fig.  and the ins Fig. , but Ad-Ad-p21 infection increased the mRNA expression of lipolytic genes in WAT and the hepatic fatty acid utilization by enhancing the mitochondrial and peroxisomal pathways of fatty acid oxidation Fig. . Ad-p21 44, and indeed Ad-p21 infection caused lower levels of phospho-S6 Ser240/244 in response to fasting  stabilizes p21, whereas p21 is destabilized by glycogen synthase kinase (GSK)3β-mediated phosphorylation at Thr5750. However, Thr145 phosphorylation by AKT/PKB does not affect p21 stability but causes its cytoplasmic translocation51. Similarly, Ser153 phosphorylation by dual specificity tyrosine-phosphorylation-regulated kinase 1B (Dyrk1B) stimulates the translocation of p21 from the nucleus to the cytoplasm53. The present study has shown that MPK38 is capable of inducing greater stability and nuclear translocation of p21 through Thr55 phosphorylation. By contrast, the stability and subcellular localization of p21 are not affected by CDK and c-Jun N-terminal kinase (JNK)/p38-mediated Ser130 phosphorylation54.p21 is known to be an unstable protein, having a half-life of about 30 min, and is degraded proteasomally in both an ubiquitin-dependent and an ubiquitin-independent mannerp21 Fig. . Recent 56. PPARγ is considered a master regulator of adipogenesis from both in vitro and in vivo studies. Indeed, PPARγ is required for adipocyte differentiation58, and in many cases its expression is sufficient for the differentiation of non-adipose cells into adipocyte-like cells60. PPARγ also regulates insulin sensitivity, lipogenesis, and adipocyte survival and function61. Thus, it is reasonable proposition that p21, a transcriptional regulator, could regulate adipocyte differentiation by affecting transactivation by PPARγ. In the present study, we found that MPK38 plays a critical role in the association between p21 and PPARγ, following Thr55 phosphorylation of p21. Indeed, phosphorylated p21 strongly interacted with PPARγ in the nucleus, leading to inhibition of PPARγ binding to peroxisome proliferator response elements (PPRE) in target genes  antibody was raised against the synthetic phosphopeptide FDFVTETPL, in which T represents phosphothreonine , in a rabbit. The WWP-Luciferase (p21/WAF1 promoter) plasmid containing the p53-binding site was from Addgene (no. 16451). MEFp21−/− cells were generated after timed matings of homozygous p21 and MEFMPK38−/− has been described previously42. The oligonucleotides were from Bioneer Ltd . Biochemical analyses, including co-immunoprecipitation, immunoblot analysis, luciferase assay, and in vitro kinase assay for MPK38, as well as quantitative real-time PCR (qPCR), confocal microscopy, and assays for apoptosis and cell cycle arrest, were performed using the indicated cells and experimental conditions, as previously described42.The antibodies, plasmids, and chemicals used have been described previouslyGAATTCATGTCAGAACCGGCTGGG-3’; reverse, 5’-GCCTCGAGTTAGGGCTTCCTCTTGGA-3’; EcoRI/XhoI site underlined), together with one of the following pairs of primer sequences: for S116A, sense 5’-GTGGACCTGTCACTGGCTTGTACCCTTGTGCCT-3’, antisense 5’-AGGCACAAGGGTACAAGCCAGTGACAGGTCCAC-3’; for S153A, sense 5’-ACAGATTTCTACCACGCCAAACGCCGGCTGATC-3’, antisense 5’-GATCAGCCGGCGTTTGGCGTGGTAGAAATCTGT-3’; and for T55A, sense 5’-AACTTCGACTTTGTCGCCGAGACACCACTGGAG-3’, antisense 5’-CTCCAGTGGTGTCTCGGCGACAAAGTCGAAGTT-3’. The PCR products were cut with EcoRI and XhoI and then subcloned into pGEX4T-1 vector using EcoRI/XhoI site to generate pGEX4T-1-p21 substitution mutants.To achieve three p21 mutants , wild-type p21 was used as the template for PCR with either the p21 forward or reverse primer  RNA (5’-AATGGCGGGCTGCATCCAGG-3’) was designed to target the genome adjacent to the p21 mutation site. The annealed oligonucleotides (5’-CACCGAATGGCGGGCTGCATCCAGG-3’ and 5’-AAACCCTGGATGCAGCCCGCCATTC-3’) containing the p21 guide sequence and Bbs1 ligation adapters were ligated into pX458 vector (Addgene) using 5 μl of 2 × Quick-Ligation Buffer and 1 μl of QuickLigase . 3T3-L1 cells were co-transfected with 1 μg p21 sg RNA plasmid and pUC19 p21 T55A to generate the p21 knock-in (T55A) cell line. The clones obtained were then verified by DNA sequencing.Genome editing with the CRISPR/Cas9 system was performed in 3T3-L1 cells, as previously describedhttps://bioscience.lonza.com/lonza_bs/US/en/document/download/21296). The differentiation of 3T3-L1 cells was induced as previously described64. Briefly, 3T3-L1 cell differentiation was induced by treatment with differentiation medium (0.5 mM 3-isobutyl-1-methylzanthine (Sigma), 1 μM dexamethasone (Sigma), and 5 μg/ml insulin ) in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS)) for 2 days. Culture medium was refreshed every 2 days with DMEM supplemented with 10% FBS and 5 μg/ml insulin. At day 10–14 of induction, fixed cells  were stained with the Oil Red O dye (Chemicon). Quantification of Oil Red O staining was achieved by measuring the absorbance at 520 nm. The isolation of hepatocytes and adipocytes was as previously described62.3T3-L1 preadipocytes were transfected with the indicated plasmids using Amaxa Nucleofector technology  according to manufacturers’ protocol (9 plaque-forming units), as previously described9.All animal experiments were approved by the Chungbuk National University, and were conducted in accordance with the approved protocols and guidelines established by the Ethics Review Committee (CBNUA-966-16-02). Male C57BL/6 or p21-null mice (4-week-old) purchased from Jackson Laboratory were placed on a high-fat diet (HFD)  for 8–10 weeks. They were kept under a 12 h light/dark cycle, with free access to autoclaved food and water, in specific pathogen-free (SPF) animal center. To prepare recombinant adenoviruses expressing wild-type (WT) and T55A p21, FLAG-tagged p21 plasmids (WT and T55A) were used as templates for PCR using the primers (5’-GTAACTATAACGGTCATGTCAGAACCGGCTGGGGATGTC-3’ and 5’-ATTACCTCTTTCTCCTTAGGGCTTCCTCTTGGAGAAGAT-3’) and the tail vein or epididymal fat pads of the HFD-fed mice were injected with adenovirus  using an ELISA kit  and an Accu-Check glucometer (Roche), respectively42. 2-deoxy-glucose uptake, ketone concentration, fatty acid oxidation rate, lipogenesis, and lipolysis were measured in HFD-fed mice infected with the indicated adenoviruses, as previously described65.Blood obtained from a tail vein was used to measure fed and fasted serum insulin and glucose in 63.Blood samples were obtained from the abdominal aorta of 18 h-fasted HFD-fed mice infected with adenoviruses expressing GFP, WT p21, T55A p21, or MPK38. To separate the serum, whole blood samples were centrifuged at 2500 rpm at 4 °C for 15 min. An ELISA kit from R&D Systems (Joslin DERC Assay Core) was used to measure serum free fatty acid concentration. An automated serum analyzer  was used to measure serum glucose, triglyceride, total cholesterol, HDL-cholesterol, and LDL-cholesterol. A triglyceride determination kit (Sigma) was used to measure serum triglyceride concentration. Serum concentrations of TNF-α, IL-6, IL-1β, and MCP1 were measured as previously describedGTTs/ITTs were performed in fasted  HFD-fed mice infected with the indicated adenoviruses. D-glucose (2 g/kg body mass) or insulin ; human R-insulin; Eli Lilly) was administrated intraperitoneally and blood glucose was measured from tail bleeds with an Accu-Check glucometer (Roche) at indicated time points after the administration.The data represent the means ± SEM of at least three independent experiments, and all experiments were performed in duplicate. Statistical analysis  was performed using one-way or two-way ANOVA with Tukey’s multiple comparison test.Supplementary Figure LegendsSupplementary Figure 1Supplementary Figure 2Supplementary Figure 3Supplementary Figure 4Supplementary Figure 5Supplementary Figure 6Supplementary Figure 7


Recent studies identified the benzoxaborole moiety as a new zinc-binding group able to interact with carbonic anhydrase (CA) active site. Here, we report a structural analysis of benzoxaboroles containing urea/thiourea groups, showing that these molecules are very versatile since they can bind the enzyme assuming different binding conformations and coordination geometries of the catalytic zinc ion. In addition, theoretical calculations of binding free energy were performed highlighting the key role of specific residues for protein-inhibitor recognition. Overall, these data are very useful for the development of new inhibitors with higher selectivity and efficacy for various CAs. Intereseriments , whereaseriments 24. The s2a, 2b, 3a, and 3b were prepared by the soaking technique3a and hCA II/3b adducts, whereas a very long soaking time was necessary for hCA II/2a and hCA II/2b (1 week for both cases). Crystals were frozen in a gaseous nitrogen stream prior to the diffraction experiment without further manipulations.Crystals of homemade hCA II in complex with ,2a, hCA II/2b, hCA II/3a, and hCA II/3b, respectively).A complete X-ray data set was collected at 100 K by a copper rotating anode generator developed by Rigaku and equipped with a Rigaku Saturn CCD detector. Diffraction data were processed and scaled using program HKL2000 2a, 2b, 3a, and 3b as well as on two model complexes with 3a* and 3b*. The latter models were obtained by substituting the oxygen atom of urea moiety with a sulphur atom in the hCA II/2a-b crystal structure, using the Builder module of Insight II package . The partial atomic charges for ligands were obtained by quantum mechanical (QM) calculations (B3LYP/6–31G*) using the Gaussian16 software ,2+ ion were adopted from the work of Li and Merzσ = 1.271; ɛ  = 0.00330286). Since boron atom is not parametrised in Amber-derived force fields, it was substituted with carbon, as reported in other modelling studies,Theoretical calculations were carried out on the four crystallographic complexes of hCA II with Gbind in kcal/mol) for crystallographic complexes and the theoretical models were calculated using the molecular mechanics/generalised Born surface area (MM/GBSA) method,per-residue basis.The binding free energies  and electrostatic (ΔEelec) contributions; ΔGsol is the solvation energy, and includes electrostatic (ΔGGB) and nonpolar (ΔGSA) interactions. The electrostatic solvation energy (ΔGGB) is evaluated using the Generalised Born methodTΔS is the change of conformational entropy on ligand binding, which is not included in our calculations. Indeed, for comparison of similar ligands, it is acceptable to exclude the entropy contribution in practise,According to MM/GBSA method, the binding free energy was estimated as follows:2a, 2b, 3a, and 3b   see , their aone) see . One of a and 3b . A questtrans-trans and trans-cis conformations depending on the nature of the phenyl substituents and on the chemical environment (data not shown). Thus, the different binding geometries assumed by urea and thiourea derivatives within the hCA II active site are induced exclusively by specific interactions with the enzyme residues. Based on the inhibition data reported in 2a and 2b is energetically more favoured with respect to that adopted by compounds 3a and 3b. Thus, to understand why thiourea-containing inhibitors do not assume the same conformation of urea-containing ones, binding free energy calculations were performed by using the MM/GBSA theoretical method,3a* and hCA II/3b*) obtained by substituting the oxygen atom of urea moiety with a sulphur atom in the hCA II/2a and hCA II/2b crystal structures. The latter two structures represent hypothetical models in which thiourea derivatives would adopt the same binding conformations observed for the corresponding urea derivatives.To answer this question, a detailed analysis of urea and thiourea bis-phenyl derivatives contained in the Cambridge Structural Database (CSD)2+3b adduct, due to a hydrogen bond between the sulphur atom of thiourea moiety and Gln92 side chain, which is absent in the case of the other derivatives  interactions . These rivatives .3a* and 3b*, respectively. These data indicate that thiourea derivatives would experience a significant destabilising interaction with Phe131 if they would adopt the same binding conformation of urea derivatives, likely due to an increase of the steric hindrance between their CS group and the Phe131 ring. These data suggest that the thiourea moiety turns towards a trans-cis conformation, to avoid this unfavourable interaction.Regarding the model complexes , the ene,,Our energetic calculations can help also to rationalise the interesting inhibition profile of the thiourea derivatives against the different hCA isoforms, with a particular focus on hCA IX and XII which have been recently recognised as valuable targets for cancer treatment and diagnosisAltogether, studies reported here demonstrate that ureido- and thioureido-benzoxaborole derivatives are an interesting class of versatile CAIs. Indeed, these molecules can bind the enzyme active site assuming different binding conformations and coordination geometries depending on the interactions established with the enzyme active site residues. This feature can be efficiently used to design molecules with enhanced selectivity for pharmaceutically relevant hCA isoforms.


Recently, with the increasing demand for virtual reality (VR), experiencing immersive contents with VR has become easier. However, a tremendous amount of calculation and bandwidth is required when processing 360 videos. Moreover, additional information such as the depth of the video is required to enjoy stereoscopic 360 contents. Therefore, this paper proposes an efficient method of streaming high-quality 360 videos. To reduce the bandwidth when streaming and synthesizing the 3DoF+ 360 videos, which supports limited movements of the user, a proper down-sampling ratio and quantization parameter are offered from the analysis of the graph between bitrate and peak signal-to-noise ratio. High-efficiency video coding (HEVC) is used to encode and decode the 360 videos, and the view synthesizer produces the video of intermediate view, providing the user with an immersive experience. As the virtual reality (VR) market is expanding rapidly, the need for efficient immersive VR technology has become more important. To play high-quality VR video through a head-mounted display (HMD), the minimum resolution of the video must be 4K. In this case, the amount of data to be processed from the HMD increases rapidly. Therefore, the Moving Picture Experts Group (MPEG) proposed a technology, which processes the viewport of the user, called motion-constrained tile set (MCTS)  in 2016;To support immersive media, the MPEG-I group, established by MPEG, divided the standardization associated with VR into three phases, namely three degrees of freedom (3DoF), three degrees of freedom plus (3DoF+), and six degrees of freedom 6DoF) . In 3DoFDoF . In A large amount of bandwidth is required for transmitting 3DoF+ or 6DoF 360 videos because such videos need both high-resolution texture and depth. To overcome this problem, down-sampling or region-wise packing could be applied. In this paper, we propose the View Location-based Asymmetric Down-sampling for Vie Synthesis (VLADVS) concept for the bitrate decreasing system with appropriate down-sampling ratio and a quantization parameter for 3DoF+ texture and depth in view synthesis, as shown in This paper is organized as follows: During the 116th MPEG meeting, the MPEG-I group was established for the support of immersive media. They began work by standardizing the format of immersive, omnidirectional video in 2017 . Figure To overcome the limitations of 3DoF, the concept of 3DoF+, part of phase 1b in MPEG-I, was proposed. 3DoF+ provides limited movements of yaw, pitch, and roll, as described in In 3DoF+, the VR device must offer the video of view that the user watches. If this video of view is not included in the original video, 3DoF+ system synthesizes the view that did not exist before. Thus, Reference Intermediate View Synthesizer  is requiAs the solutions to the abovementioned problems, enhanced communication technologies such as 5G mobile technology  and mobiX-, Y-, and Z-axes. It supports the user’s movements including walking, as described in View synthesis assumes video transmission from the server to the client. Therefore, the video must be compressed, as shown in Multi-view video provides the user with an immersive 3D experience. Such video provides diverse views gained from one scene simultaneously. Particularly, 3D multi-view video includes both texture and depth information. It enables users to have multiple views of what they intend to watch. MPEG defined a 3D video system , which iMulti-view videos have common features as they contain the same scene at the same time. The difference between each view is the indigenous point of view; that is, a multi-view video of one viewpoint can be made by referencing another view.Although multi-view video provides some views, it cannot offer out-of-source views. Because multi-view video coding requires a large amount of data and computing power to process, the number of views the multi-view video can support is limited. Accordingly, view synthesis for multi-view video ,27 was dX-, Y-, and Z-axes. The X-, Y-, and Z-axes represent the horizontal, vertical, and optical axis , respectively. The optical axis is the direction of the camera ray. The principal point is the intersection point between the principal axis and the image plane. The distance from the camera center to the principal is called focal length, as shown in Generally, the multi-view video is obtained from a pinhole camera. It projects the actual object onto a 2D plane image, as shown in To obtain the intermediate view, the point coordinates from reference views must be converted into the synthesized view. Each reference view, which is used to synthesize the intermediate view, has its own camera coordinate system. If we realize the camera parameter of reference views and intermediate view, the camera coordinate system of intermediate view can be generated using the world coordinate system. Once the conversion is complete, texture mapping from the reference views to intermediate view is performed.This section explains VLADVS for efficient use of bandwidth in video transmission, as described in To reduce the bitrate when transmitting multi-view video, this paper proposes a low-complexity multi-view video transmit system including down-sampling and up-sampling. The feasibility of this method was proved by a pilot test with FTV multi-view sequences .Champagne_Tower and Pantomime sequences, as shown in For encoding, the quantization parameter (QP) values are 22, 27, 32, and 37. In a pilot test with FTV multi-view sequences, the experiment was executed for every combination of down-sampling ratio, QP, and view synthesis. The pilot test results are shown in For the 3DoF+ experiment, MPEG provides Classroom-Video , TechnicIn another experiment, view v0 was defined as a synthesized view; v1, v2, v3, v4, v5, and v6 were called near views; and v9, v10, v11, v12, v13, v14 were called far views, as shown in In In CTC, the QPs used for texture and depth are shown in The goal of this experiment is to reduce the bitrate while conserving the PSNR. Modifying parameters such as down-sampling ratio, QP, and the number of input views to optimize them are included in the experiment, which is explained in In X-axis were QP of R1–R4. R2 can be used instead of R1; the gap between R1 and R2 was not high. With QP of R2 and 12.5% down-sampling ratio, it saved approximately 87.81% bitrate while losing only 8% WS-PSNR, compared to the result of R1 and 0% down-sampling ratio.In addition, experiment with two down-sampling ratios was conducted. After sorting the source views by the distance between the source views and intermediate views, the experiment assigned two down-sampling ratios to the source views. If the source views are close to the intermediate view, they got low down-sampling ratios. To decide the combination of two down-sampling ratios, the following formula is used:Here, n is the number of the entire down-sampling ratios, and r is the number of the down-sampling ratios to assign. To obtain the number of DR1 and DR2 to the source views, the following equations are used:Equation (2) explains how to calculate the number of DR1. After dividing the number of source views with 2, which means the number of down-sampling ratios to assign, the formula rounds up the result. DR2 is set to the difference value between the number of source views and the number of DR1, as shown in Equation (3).In This paper proposes a bitrate-reducing method for 3DoF+ video synthesis and transmission. Particularly, by down-sampling and up-sampling the texture and depth, the proposed method saves the bitrates of bitstream file while degrading the objective video quality very little in WS-PSNR. In addition, down-sampling the far views brings higher WS-PSNR value than down-sampling all the source views. However, because the number of the parameters for the experiment was not enough to deduct the optimal parameter for view synthesis, the experiment using video compression methods such as region-wise packing  must be


The differing glycemic response to trifle was not related to satiety or to subsequent energy intake.The effect that blood glucose concentration has on feelings of satiety is unclear. Our aims were to assess satiety and subsequent energy intake following the ingestion of trifle sweetened with sucrose or isomaltulose whilst measuring plasma glucose concentration to confirm glycemic differences between trifles. Seventy-seven healthy adults participated in a double-blind crossover trial where trifle sweetened with sucrose or isomaltulose was consumed on separate days with a two-week washout. Blood was sampled at the baseline, 1 and 2 h postprandially, and satiety assessed using visual analogue scales (VAS). Weighed diet records were taken on test days. A statistically significant difference in blood glucose concentration between trifles was found at 60 min following consumption, with the isomaltulose trifle having a 0.69 mmol/L  lower concentration when compared with the sucrose trifle. Mean satiety response by area-under-the-curve (AUC) was not significantly different between trifles. Mean (SD) appetite scores for the sucrose and isomaltulose trifles were 4493 (2393) and 4527 (2590) mm·min, respectively, with a between trifle difference of −9  mm·min. Mean (SD) energy intake for the remainder of the day following trifle consumption was 3894 kJ (1950 kJ) and 3530 kJ (1926 kJ) after the sucrose and isomaltulose trifles, respectively, and was not significantly different ( Overweight and obesity occur on a global scale and efforts are needed to counteract the problems of obesity-related diseases . The WorHowever, any effects of circulating blood glucose on satiety are unclear, as findings have been inconsistent . Part ofThe problem of attributing satiogenic effects to the glycemic response properties of foods selected on the basis of GI, independent of other properties of the food, has been reviewed . The autAnother strategy to manipulate glycemic responses by exchanging food ingredients is to use sugars with different GI. Isomaltulose  is a non-cariogenic sugar found in trace amounts in honey  and in JThe objective of this experiment was to compare the acute effect on satiety by incorporating a higher and a lower GI sugar as an ingredient into a solid food that could be consumed in practice. In order to test the hypothesis that feelings of satiety would be increased following the consumption of a food containing a lower GI ingredient, it was necessary to identify a food with a relatively high sugar content so that it would generate a difference in glycemic response between test foods. Trifle was chosen because each of its components  contain a considerable proportion of sugar. Hence, the current study was designed to measure glycemic responses and to test for short-term effects on satiety and subsequent energy intake by providing participants with trifle sweetened with either isomaltulose or sucrose. The main outcomes were satiety and subsequent energy intake, with plasma glucose concentration measured as confirmation of the effectiveness in generating glycemic differences between trifles.Participants were a convenience sample of students from the University of Otago. Inclusion criteria were students enrolled in a third year human nutrition course older than 18 years of age. Students were invited to participate in the study providing they had no food allergies to any of the trifle ingredients. Students were not obliged to take part and were given an information sheet and the opportunity to seek clarification of what the study involved. The University of Otago Human Ethics Committee approved the study (reference H17/011) and students signed a consent form. The study has been registered with the Australian and New Zealand Clinical Trials Registry ACTRN12618001137280.Seventy-seven young adults received a sucrose- or isomaltulose-sweetened trifle at lunchtime in a cross-over design randomized to the order in which they received the trifle. Participants attended two testing days on Fridays starting at 12 p.m. with a 2-week washout. Participants were stratified by the weeks when they could attend the test sessions to ensure equal distribution at each clinic for the order in which the trifles were consumed. The day before the first test day, each participant indicated to the investigators the type and amount of cereal that he or she wished to consume at breakfast the following morning and this was weighed and packaged in a sealable plastic bag for the participant to take home. Participants were instructed to eat all of the cereal on the morning of the test day at his or her usual breakfast time and then not to eat or drink (apart from water) until 12 p.m. Participants were provided with the same breakfast and asked to eat the breakfast at the same time on each of those days to ensure that appetite and energy intake prior to the lunchtime test sessions were consistent.At 12 p.m., participants were seated in the testing facility and asked to consume the trifle within 20 min. No other food was consumed for the following 150 min. A staff member independent of the study used the random number generator in Excel  to allocate the order in which each student received the sucrose- or isomaltulose-sweetened trifle. This staff member placed a colored sticker (red or green) onto the lid of the trifle container corresponding to sucrose or isomaltulose. The students, study investigators, and the biostatistician were blinded to trifle type with the colored code revealed after the completion of the statistical analysis. On the morning of the first test day, the participants’ heights were recorded to the nearest mm using a stadiometer ; and weight was measured to the nearest gram using calibrated electronic scales . Body mass index was calculated as weight divided by height squared. Participants filled out a questionnaire regarding sex, age, and ethnicity.The trifles were made in the metabolic kitchen of the Department of Human Nutrition at the University of Otago. The ingredients were: eggs, sugar , plain flour, cornflour, baking powder, water, lemon juice, gelatin, full fat milk, and cream. Each serving weighed 446 g and contained 2600 kJ. The nutritional composition of the two trifles on a fresh weight basis were identical: protein 15.8 g; fat 18.6 g; available carbohydrate 98.2 g; total sugars 80.5 g ; dietary fiber 0.6 g, and ash 2.1 g; with a moisture content of 70%.g within 20 min of blood collection and the plasma was extracted and stored at −80 °C until analysis. Plasma glucose concentration was measured using the glucose hexokinase method on a Cobas c311 auto analyzer . Coefficients of variation for Roche control sera Precinorm U plus  and Precipath U plus  were 1.25% and 0.67%, respectively. A 500 µL capillary blood sample was collected into a microcontainer containing potassium ethylenediaminetetraacetic acid (EDTA)  using a contact-activated disposable lancet at the baseline and at 1 and 2 h following consumption of the trifles. The tubes were centrifuged for 10 min at 2000× Feelings of satiety were assessed using 100 mm Visual Analogue Scales (VAS) using methodology validated by Flint and colleagues . Four quTraining was given to participants in the use of the Model 3010 Salter electronic kitchen scales reading to 1 g . Participants took the scales home and weighed and recorded all food and beverages consumed throughout the day from waking on the morning of each test day through to midnight. The dietary data were entered into a University of Otago dietary analysis program that uses the New Zealand Food Composition database as the source of nutrient information .p-value < 0.05 was considered statistically significant. Stata 15.1  was used to analyze the VAS satiety data.A sample of 60 was required to detect a difference of 0.5 standard deviations for all outcomes in standardized form with 90% power and α = 0.01. Seventy-seven participants were recruited as a convenience sample, which allowed for some dropout. Random effects regression analysis was used to test for between-treatment differences in plasma glucose at the 60 and 120 min timepoints and for AUC satiety responses with participant id as a random effect and adjusted for randomized order and baseline satiety. Analysis was also undertaken for standardized AUC and to estimate differences in subsequent energy intake. A Seventy-seven participants were randomized to order and complete blood and satiety data were available for 66 people. A diagram of participant flow through the study is given in Anthropometric and demographic characteristics of the sample are given in Participants were mainly young female adults of European descent, with Asian and Māori ethnicities combined constituting 30% of the sample.p = 0.253). The mean blood glucose concentration data sampled at 60 and 120 min and comparisons between treatments at 60 and 120 min are given in The mean (SD) plasma glucose concentrations at the baseline were 5.2 (0.7) and 5.1 (0.7) mmol/L for the sucrose and isomaltulose-sweetened trifles, respectively, and these concentrations were not different  to 150 min postprandial. This enabled six VAS questionnaires to be completed by each participant on a given testing day. There were no significant differences for mean difference between the isomaltulose- and sucrose-sweetened trifle or in mean AUC for each satiety question across all time points following the consumption of the trifles .p = 0.133.A meaningful effect was ruled out as the mean difference and 95% CI were all under 0.3 standard deviations. Therefore, it is unlikely that there is a real difference in satiety between the trifles. Sixty-six participants recorded subsequent energy intake for the rest of the day after the trial; mean (SD) energy intake after the sucrose trifle was 3894 kJ (1950 kJ), and after the isomaltulose trifle it was 3530 kJ (1926 kJ). Energy intake for the remainder of the day after consuming the trifle did not differ significantly between treatments with a mean difference  of 364 kJ , In the present study, differences in postprandial glycemia were found between trifles, but there were no significant differences in the participants feelings of satiety or in their subsequent energy intake. These findings are consistent with other work. When comparing satiety among 38 foods, food volume or energy density were found to be the strongest predictors of satiety index scores, with satiety index defined as area under the 120 min satiety curve (AUC) of the test food divided by the AUC of white bread . Using tIn contrast, differences in some appetite measures have been found from studies where comparison treatments have been designed using food choices based on GI. In one such study, food was requested approximately three-quarters of an hour earlier after the high GI meal when compared with the low GI meals, although there was no difference in subsequent energy intake . In a stThe outcomes of these studies are variable both within and among studies, but in each study there was some indication that low, as opposed to high GI foods, resulted in some greater measure of satiety. However, whether any of the differences found were due to glycemic responses is uncertain. The fiber content of foods has been found in some studies to affect satiety . In the Controlling for factors such as fiber, energy, and macronutrient content is possible with the use of beverages sweetened with sugars with different glycemic-inducing characteristics. Beverages sweetened with glucose (G) and fructose (F) mixtures of G80:F20 (high glycemic) and G20:F80 (low glycemic) ratios were given to 12 and 19 people (two experiments), resulting in no difference in the ratings of appetite but a lower subsequent energy intake 80 min after drinking the high-compared with the low-glycemic beverage . In anotA means of isolating the glycemic effects of sugars on satiety is the use of isomaltulose and sucrose as the comparison treatments as these two sugars have identical monosaccharide constituents. It has been found that rats provided with sucrose or isomaltulose ingested more energy over 24 h when exposed to sucrose compared with isomaltulose . HoweverA major strength of the study was the use of isomaltulose and sucrose as the sweeteners that allowed for double-blinding and for the control of many factors associated with satiety including volume of food, macronutrients, fiber, and energy content. The study also had a strong design, being a crossover, participants were randomized to treatment order, and it was adequately powered with a relatively large sample. A limitation was the inclusion of these sugars into trifle that limited the glycemic difference between treatments. A maximum difference of around 1.5 mmol/L in blood glucose concentration between sucrose and isomaltulose has been found when participants ingested 50 g solutions of these beverages  whereas In conclusion, sucrose and isomaltulose contain identical glucose and fructose molecules but differ in the glycosidic bond joining the monosaccharides, resulting in the slower digestion of isomaltulose when compared with sucrose. The slower rate of digestion of isomaltulose compared with sucrose generated a glycemic difference between the two trifles at lunchtime, but this glycemic difference did not result in differences in feelings of satiety or in subsequent food intake over the remainder of the day. These data are novel and will hopefully lead to other investigators testing the satiating properties of these sugars amongst a wider demographic.


Fusarium cause Fusarium head blight (FHB) of cereal crops and contaminate grain with sesquiterpenoid mycotoxins, including culmorin (CUL) and trichothecenes. While the phytotoxicity of trichothecenes, such as deoxynivalenol (DON), and their role in virulence are well characterized, less is known about the phytotoxicity of CUL and its role in the development of FHB. Herein, we evaluated the phytotoxic effects of purified CUL and CUL-trichothecene mixtures using Chlamydomonas reinhardtii growth and Triticum aestivum (wheat) root elongation assays. By itself, CUL did not affect growth in either system. However, mixtures of CUL with DON, 3-acetyldeoxynivalenol, 15-acetyldeoxynivalenol, or NX-3, but not with nivalenol, inhibited growth in a synergistic manner. Synergistic phytotoxic effects of CUL and DON were also observed on multiple plant varieties and species. The severity of wheat FHB caused by 15 isolates of Fusarium graminearum was negatively correlated with the CUL/DON ratio, but positively correlated with the sum of both CUL and DON. Additionally, during the first week of infection, CUL biosynthetic genes were more highly expressed than the TRI5 trichothecene biosynthetic gene. Furthermore, genomic analysis of Fusarium species revealed that CUL and trichothecene biosynthetic genes consistently co-occur among species closely related to F. graminearum.Species of the fungus  Fusarium species, such as Fusarium graminearum, produce an array of secondary metabolites, including mycotoxins, that can pose serious risks to plant, animal, and human health .,49.Fusar queries ,14,50,51Fusarium, species trees were inferred from nucleotide sequences of 11 primary-metabolism genes that have been used previously for species tree inference in Fusarium and other fungi (CLM1 and CLM2 were also aligned with MEGA7, and the resulting alignments for each gene were subjected to maximum likelihood analysis with IQ-Tree as described above.To map the presence and absence of the biosynthetic genes to er fungi  [35,37,5


In T2D patients, treatment with metformin is associated with reduced blood Mg2+ levels. To investigate how T2D and metformin affect Mg2+ homeostasis db/m and db/db mice were treated with metformin or placebo. Mice were housed in metabolic cages to measure food and water intake, and to collect urine and feces. Serum and urinary Mg2+ concentrations were determined and mRNA expression of magnesiotropic genes was determined in kidney and distal colon using RT-qPCR. Db/db mice had significantly lower serum Mg2+ levels than db/m mice. Mild hypermagnesuria was observed in the db/db mice at two weeks, but not at four weeks. Metformin-treatment had no effect on the serum Mg2+ concentration and on the urinary Mg2+ excretion. Both in kidney and distal colon of db/db mice, there was a compensatory upregulation in the mRNA expression of magnesiotropic genes, such as transient receptor potential melastatin 6 (Trpm6), whereas metformin treatment did not affect gene expression levels. In conclusion, we show that T2D causes hypomagnesemia and that metformin treatment has no effect on Mg2+ homeostasis in mice.Approximately 30% of patients with type 2 diabetes mellitus (T2D) have hypomagnesemia (blood magnesium (Mg Hypomagnesemia has serious clinical consequences as it increases the risk of complications such as retinopathy, nephropathy, micro and macrovascular disease and foot ulceration4. Moreover, Mg2+ deficiency is correlated with insulin resistance, abrogated glucose metabolism and an increased risk of developing T2D7. However, the etiology and underlying mechanisms of hypomagnesemia in T2D patients remains largely unknown8.Approximately 30% of patients with type 2 diabetes mellitus (T2D) have hypomagnesemia  transport11. In the colon, the final absorption of Mg2+ takes place by an active transcellular mechanism through transient receptor potential melastatin type 6/7 (TRPM6/TRPM7) cation channels9. In the kidney, 95–99% of filtered Mg2+ is reabsorbed under physiological circumstances9. Approximately 85% of the filtered Mg2+ is reabsorbed paracellularly by the proximal tubule and the thick ascending loop of Henle (TAL), where transport relies on tight junction permeability14. Active transport in the distal convoluted tubule (DCT) determines the final urinary Mg2+ concentration, as this is the final segment where Mg2+ is reabsorbed15. In physiological conditions, the DCT reclaims 5–10% of filtered Mg2+ transcellularly via TRPM6/7 channels16. The expression and/or the activity of TRPM6 is affected by SNPs, dietary Mg2+ intake, drugs and hormones, such as insulin and epidermal growth factor (EGF)20. SNPs in TRPM6 that impair its response to insulin have been associated with an increased risk of developing T2D and gestational diabetes19.The intestine and kidney collaboratively regulate Mg21, suppresses hepatic gluconeogenesis and improves insulin sensitivity22. Therefore, its major clinical benefit is reducing blood glucose levels with only a minimal risk of hypoglycemia24. The most common side effects of metformin treatment are lactic acidosis, nausea and diarrhea25. Recent cohort studies showed that metformin use in T2D patients is associated with reduced blood Mg2+ levels26. However, the mechanism that underlies this correlation has not yet been elucidated. To investigate how T2D and metformin affect Mg2+ homeostasis, control (db/m) and diabetic (db/db) mice were treated with placebo or metformin for four weeks. Serum and urinary electrolytes were measured and mRNA expression of magnesiotropic genes was evaluated in kidney and distal colon.Metformin, the first-line pharmacotherapy in T2Dad libitum access to tap water and standard pellet chow. At day 0, diets were changed to a diet containing 0.05% (w/w) MgO  and drinking water to demineralized water. At days-2, 12 and 26 mice were housed individually in metabolic cages for 48 hours  to measure food and water intake and to collect urine and feces. Mice were weighed twice weekly and blood was collected via the submandibular vein at days -2 and 15. Mice were randomly divided into four experimental groups of ten mice per group, of which half received metformin hydrochloride , dispersed in the drinking water. Researchers and animal caretakers were blinded for the metformin treatment. After 28 days of treatment, mice were anaesthetized by 4% (v/v) isoflurane and exsanguinated by orbital sinus bleeding, and death was confirmed by cervical dislocation. Colon and kidney tissues were cleaned with ice-cold PBS and snap-frozen in liquid nitrogen.The animal study was approved by the animal ethics board of the Radboud University Nijmegen (RU DEC 2015-0073) and by the Dutch Central Commission for Animal Experiments . Experimental procedures were conducted in accordance with the institutional guidelines and in compliance with Dutch and European laws and policies. Twenty diabetic (db/db) and twenty control (db/m) male mice , aged 8–10 weeks, were acclimatized for two weeks in a temperature- and light-controlled room two mice per cage (Eurostandard Type IIL), with Hprt), the relative gene expression was calculated by the Livak method (2−∆∆ct). Primers sequences are provided in Table 1.TRIzol reagent  was used to extract total RNA from kidney and distal colon according to the manufacturer’s protocol. RNA was subjected to DNase  treatment at 37 °C for 30 min and then to DNase stop buffer at 65 °C for 10 min. The RNA concentration was measured using the Nanodrop 2000c . To synthetize cDNA, 1.5 µg of total RNA was reverse transcribed for 1 hour at 37 °C using Moloney-Murine Leukemia Virus (M-MLV) reverse transcriptase . SYBR Green Supermix  was used to analyze the gene expression levels on a BioRad  analyzer. After normalizing to housekeeping gene expression , according to manufacturer’s protocol. Ca2+ concentrations were determined by the o-cresophthalein complexone method. Absorbance for the Mg2+ and Ca2+ assays was measured at 600 nm and 570 nm, respectively, on a Bio-Rad Benchmark plus microplate spectrophotometer . Serum and urinary Na+ and K+ concentrations were measured at the clinical chemistry department applying standardized methods1. Serum and urinary glucose concentrations were determined by a spectrophotometric assay according to the manufacturer’s protocol .Serum and urinary Mgt test, with the Holm–Sidak method for multiple comparisons, was used. In the absence of a significant interaction effect, a two-way ANOVA approach with a Tukey’s multiple comparisons test was used. Statistical significance was assessed using Graphpad Prism v7  was investigated using a two-way ANOVA test. If there was a significant interaction effect, an unpaired multiple 2+ homeostasis, control (db/m) and diabetic (db/db) mice were treated with metformin or placebo for four weeks. Db/db mice were significantly heavier than db/m mice . Metformin treatment had no effect on body weight in both db/m and db/db mice  and four weeks . At two weeks, there was a significant genotype effect on urinary Mg2+ excretion, demonstrating an increased urinary Mg2+ loss in db/db mice , whereas no significant difference was observed at four weeks . There were no significant differences on urinary Ca2+ excretion . Urinary excretion of Na+ and K+ was higher in db/db than db/m mice, and metformin treatment reduced Na+ and K+ excretion only in db/db mice .Serum Mgeks Fig. . At fourion Fig. . Despiteice Fig. . Serum K2+ levels decrease, intestinal uptake of Mg2+ is enhanced15. Colonic mRNA expression of Trpm6, the major channel for regulated Mg2+ absorption, was elevated in db/db compared to db/m mice   was lower in both db/db groups .When serum Mgice Fig. . There wm4) Fig. . The col2+ channel Trpm6, and the basolateral Mg2+ extruder Slc41a3  in db/db mice . In contrast, the mRNA expression of Cldn14, Cldn16 and Cldn19 was enhanced in db/db mice , despite major improvements in the blood glucose concentration29. In our study, metformin treatment did not affect the serum Mg2+ concentration and urinary Mg2+ excretion in db/db and db/m mice. In addition, metformin did not alter gene expression of colonic and renal Mg2+ transporters. This is in line with a study that observed no effect of a two-week metformin treatment on serum Mg2+ levels in streptozotocin-induced diabetic rats31. Possibly, a two- to four-week treatment duration is too short to detect effects on Mg2+ homeostasis. The association between metformin and lower serum Mg2+ levels in T2D patients could also be caused by other factors that were not included in the analyses. For instance, a well-known side effect of metformin-treatment is chronic diarrhea, leading to intestinal malabsorption and hypomagnesemia28.Hypomagnesemia is a common clinical feature in T2D patients35. A remaining question is whether hypomagnesemia is the cause or the consequence of T2D8. In the present study, db/db mice developed hypomagnesemia, indicating that hypomagnesemia is a consequence of T2D. At the fourth week of the experiment, db/db mice developed massive glycosuria but no renal Mg2+ wasting. This finding is against the leading hypothesis that renal Mg2+ wasting in T2D patients is a result of glycosuria36. Indeed, metformin treatment noticeably decreased glycosuria in db/db mice but did not modify the urinary Mg2+ excretion. This is in line with recent observations that glycosuria-causing SGLT2 inhibitors, lead to a mild increase in serum Mg2+ levels38. Therefore, it is unlikely that glycosuria underlies hypermagnesuria-induced hypomagnesemia in T2D. As db/db mice develop severe hyperinsulinemia, the observed hypomagnesemia could be a consequence of a Mg2+-shift towards the intracellular compartment, induced by insulin41. Future studies should focus on measuring intracellular Mg2+ concentrations in diabetic mice.Hypomagnesemia is prevalent in over 30% of T2D patients2+ concentration within the physiological range15. The DCT is the final segment where Mg2+ can be reabsorbed9. In the DCT, regulated Mg2+ reabsorption takes place transcellularly via TRPM618. Mg2+ uptake by TRPM6 is dependent on NCC, although the underlying mechanism remains largely unknown43. Gene expression levels of Trpm6 and Slc12a3, encoding for NCC, were enhanced in db/db mice, indicative of compensation in the DCT. As only a minor hypermagnesuria is observed at two-weeks, and no hypermagnesuria at four-weeks, there appears to be proper renal compensation in the db/db mice. The TAL is responsible for the bulk of renal Mg2+ reabsorption9. In the TAL, paracellular Mg2+ and Ca2+ reabsorption is regulated by the Cldn14/16/19 complex45. Cldn14 mRNA expression is strongly regulated by dietary Ca2+ intake47. The high food intake, and therefore high Ca2+ intake, of db/db mice is likely the underlying cause of the extensive upregulation of Cldn14 expression. The high expression of Cldn14 will have a negative effect on Mg2+ reabsorption in the TAL, leading to a compensatory increase in Cldn16/19 expression48. In contrast, gene expression of Cldn10b was decreased. Cldn10b enhances the Na+-permeability of the TAL, and thereby indirectly increases uptake of Mg2+ and Ca2+ in the TAL. Therefore, Cldn10b-deficient mice develop hypermagnesemia and hypomagnesuria. Likely, the observed reduction in Cldn10b expression in the db/db mice is a response to the high osmolality of the pro-urine. INSERT ENTER The strength of this study is that using oral metformin treatment in diabetic mice closely resembles the human situation. Db/db mice developed hypomagnesemia making them an excellent model to study the mechanisms of hypomagnesemia in T2D. Moreover, this study extensively investigated differences in expression of all known genes involved in Mg2+ transport, in both kidney and colon. Some limitations have to be considered. The fact that metformin treatment did not affect Mg2+ homeostasis raises the question whether the dose and duration of metformin treatment were sufficient. However, the metformin treatment reduced the food intake of db/db mice, a known positive effect of metformin. Moreover, the dosage of metformin that the db/db received  is similar to previous studies investigating the metabolic effects of metformin in mice52. A second limitation is that the expression of genes such as Cldn10b/14, Slc12a1 and Slc12a3 is regulated by both dietary intake and serum levels of K+, Na+ and Ca2+ 53. As db/db mice have hyperphagia, their dietary intake of ions is also increased. Despite the higher food intake, db/db mice still develop hypomagnesemia. However, for other differences between db/m and db/db mice it is difficult to differentiate whether they are caused by T2D-related factors or by a higher food intake.The kidneys are essential in maintaining the serum Mg2+ concentrations is likely mediated by other factors, and not by a direct effect of metformin on Mg2+ (re)absorption.In conclusion, hypomagnesemia is a consequence of T2D, which is not affected by metformin treatment. The reason that metformin-users have lower serum Mg


Additionally, the iK parameters of these molecules for BChE and AChE were calculated in the ranges 23–88 and 18–78 nM, respectively.Compounds containing nitrogen and sulfur atoms can be widely used in various fields such as industry, medicine, biotechnology and chemical technology. Therefore, the reactions of aminomethylation and alkoxymethylation of mercaptobenzothiazole, mercaptobenzoxazole and 2-aminothiazole were developed. Additionally, the alkoxymethyl derivatives of mercaptobenzoxazole and 2-aminothiazole were synthesized by a reaction with hemiformals, which are prepared by the reaction of alcohols and formaldehyde. In this study, the inhibitory effects of these molecules were investigated against acetylcholinesterase (AChE), butyrylcholinesterase (BChE) enzymes and carbonic anhydrase I, and II isoenzymes (hCA I and II). Both hCA isoenzymes were significantly inhibited by the recently synthesized molecules, with  Chemists are interested in derivatives of mercaptobenzothiazole and mercaptobenzoxazole because a number of biologically and physiologically active compounds with bactericidal, fungicidal, tuberculostatic, anti-inflammatory, parasympatholytics and anesthetic properties have been synthesized based on them.2) and water (H2O) to bicarbonate (+).The carbonic anhydrases  are a superfamily of metalloenzymes that catalyze a crucial and simple biochemical reaction, the reversible hydration of carbon dioxide  is a growing priority for pharmaceutical research and discovery. In addition to the defined role of CAIs as antiglaucoma drugs and diuretics, their potential as anti-obesity, anti-convulsant, anti-infective and anticancer has been recently described.,,,,,Alzheimer’s disease (AD) is the most prevalent cause of dementia in elderly people.1–17) and to generate more potent BChE and AChE enzymes, CA II and I isoforms.The goal of this paper is to design and synthesize some novel aminomethyl and alkoxymethyl derivatives  in ethanol. The resulting product was recrystallized from methanol. The aminomethyl derivatives of benzothiazole and benzoxazolthiones Initial aminals were obtained by condensing of secondary amines with formaldehyde. The physico-chemical characteristics of the obtained products are shown in Formaldehyde was used as a form of paraformaldehyde. The reaction was carried out in an absolute ethanol solution. Hemiformals reacted immediately after its preparation without isolation. The resulting reaction water was separated by azeotropic distillation with benzene. The crystals were obtained after distilling the solvents, including ethanol and benzene, and recrystallizing. The melting points and yields are given in To do this, hemiformal was obtained from 0.05 mol of a formaldehyde  and 40 ml of the corresponding alkanol (taken in excess as a solvent). Hemiformal reacted immediately after its preparation without isolation. Then, 0.05 mol of mercaptobenzoxazole (or 2-aminothiazole) dissolved in ethanol was added to hemiformal at the temperature of 10 °C. The resulting reaction water was separated by azeotropic distillation with benzene. The crystalline substances were obtained after distilling off the solvent  and recrystallization.1–17) on CA I, and II isoforms, which purified from fresh human erythrocyte using an affinity chromatography procedure.,et al.,,p-Nitrophenylacetate (PNA) compound was used as a substrate, and it was converted by both isoforms to p-nitrophenolate ions.,,,50 values were calculated from activity (%) against compounds inhibition.iK values.To observe of inhibition effects of novel aminomethyl and alkoxymethyl derivatives (1–17) on AChE and BChE activities were measured according to Ellman et al.The inhibitory effects of novel aminomethyl and alkoxymethyl derivatives  properties, were used as amino-methylation reagents.The structure of the products was established by NMR spectroscopy and the composition was confirmed by elemental analysis. Spectra were measured on a Bruker device in acetone. A singlet at 4.6 ppm corresponding to N-CHThe alkoxymethyl derivatives of mercaptobenzoxazole and 2-aminothiazole were synthesized by reacting them with hemiformals, which were prepared by the reaction of alcohols with formaldehyde.in vivo and in vitro; therefore new therapeutic approaches targeting either hCA IX/XII (for antitumor activity) or hCA II (for antiglaucoma action) have been developed.,iK and IC50 of the aminomethyl and alkoxymethyl derivatives (1–17) were calculated and they are given in Sulfamate and sulfonamide CAIs demonstrated fundamental anti-glaucoma and anti-tumour activities 1–17) had iK values in the range of 58 ± 15 to 157 ± 38 nM (8 (iK: 58 ± 15 nM); N-morfolinomethylbenzoxazoline-2-thion and compound 5 (iK: 70 ± 15 nM); N-diethylaminomethylbenzothiazoline-2-thione) inhibited the hCA I isoform more potently than the standard compound AZA (iK: 333 ± 28 nM), which is used to treat glaucoma, cystinuria, periodic paralysis, epileptic seizure, dural estasia and central sleep apnea. hCA I is involved in retinal edema and cerebral and the inhibition of hCA I can be a significant factor for eliminating of these conditions.Cytosolic hCA I, and II isoenzymes are widely distributed throughout the human body and interference with these enzymes may cause side effects. For the cytosolic hCA I enzyme, aminomethyl and alkoxymethyl derivatives ( ± 38 nM . Especia+) into the eye along with the influx of water, which leads to an increase in intraocular pressure.1–17) had iK values ranging from 81 ± 19–215 ± 40 nM. In addition, AZA compound applied as a standard CA inhibitor, which obtained iK value of 353 ± 60 nM. As can be observed in hCA II, the most considerable inhibition result was recorded by N-oxazinomethylbenzothiazoline-2-thione (1) (81 ± 19)  .1–17). It was calculated that iK values were in the range of 23 ± 3–88 ± 11 nM for BChE and 18 ± 2–78 ± 36 nM for AChE, respectively methyl-benzoxazoline-2-thione) showed the highest selectivity for AChE over BChE (ratio: 0.388) and weakest compound was 6 (N-diethylaminomethylbenzoxazoline-2-thione) (ratio 1:500).In this work, we calculated AChE/BChE selectivity. The most promising compound 1–17) with cofactor Zn2+ ions in the structure of the isoforms. For hCA I isoform  was good inhibited by entire of the evaluated molecules, the best inhibitors of them were N-diethylaminomethylbenzothiazoline-2-thione (5), N-morfolinomethylbenzoxazoline-2-thion (8) and N-oxazolidinomethylbenzoxazoline-2-thione (9) (16) and 2-(methoxy)methylaminothiazole (17) compounds are weaker inhibitors compared to other compounds for this isoenzyme. The molecule 8 was shown to had the excellent inhibitory efficacy on hCA I isoenzyme activity while the molecule 1 was shown to had the excellent inhibitory efficacy on hCA II isoenzyme activity. For hCA II isoform, the best inhibitors of them were N-oxazinomethylbenzothiazoline-2-thione (1) and N-isopropoxymethylbenzoxazoline-2-thione (13). The 2-(methoxyethoxy) methylaminothiazole (15) and 2-methoxymethylaminothiazole (17) molecules are weaker inhibitors compare with other molecules for this isoform. As seen in 50 values are in the range of 89–187 nM towards hCA II, while for hCA I is in the range of 79–156 nM. The IC50 values for standard molecule AZA towards hCA II and I are 520 and 373 nM, respectively. All molecules have lower IC50 value compare with AZA toward hCA II and hCA I isoenzymes.The synthesized molecules are shown to inhibit hCA II and I isoenzymes by the interplay of aminomethyl and alkoxymethyl derivatives (ione (9) . The 2-i50 amounts were in the range of 36–89 nM towards AChE, while they were in the range of 48–145 nM towards BChE , N-(methoxyethoxy)methyl-benzoxazoline-2-thione (14) and also for BChE were N-diethylaminomethylbenzoxazoline-2-thione (6) and N-morfolinomethylbenzoxazoline-2-thion (8), respectively.As seen in rds BChE . The IC5iK amounts were obtained for entire novel aminomethyl and alkoxymethyl derivatives (1–17) and these molecules can be considerable inhibitor of AChE, BChE enzymes and both hCA isoforms. The molecules 5 and 8 towards hCA I and molecules 1 and 13 towards hCA II and molecules 4 and 14 towards AChE and molecules 6 and 8 towards BChE enzymes recorded which can to be the leader molecules of the parts for subsequent evaluations.In this paper, nanomolar levels of


Robots performing automated tasks in uncontrolled environments need to adapt to environmental changes. Through building large collectives of robots, this robust and adaptive behavior can emerge from simple individual rules. These collectives can also be reconfigured, allowing for adaption to new tasks. Larger collectives are more robust and more capable, but the size of existing collectives is limited by the cost of individual units. In this article, we present a soft, modular robot that we have explicitly designed for manufacturability: Linbots. Linbots use multifunctional voice coils to actuate linearly, to produce audio output, and to sense touch. When used in collectives, the Linbots can communicate with neighboring Linbots allowing for isolated behavior as well as the propagation of information throughout a collective. We demonstrate that these collectives of Linbots can perform complex tasks in a scalable distributed manner, and we show transport of objects by collective peristalsis and sorting of objects by a two-dimensional array of Linbots. This interest is due to the hypothesis that multiple low-cost units are more robust than a single, advanced robot.2 Modular robot collectives can be reconfigured for different tasks,3–5 setting them apart from monolithic robots. Collectives of modular robots have greater capability than the sum of their parts.6 This increased capability allows modular collectives to be used for tasks that are not possible with monolithic robots such as reconfigurable furniture7 and modeling cell collectives.8 Although modular actuators can also be combined into a single system for tasks such as controllable surfaces,9–11 collectives of modular robots have the advantages of reconfigurability and inbuilt sensing and computation.T1 The size of a collective robotic system, however, is often limited by cost, with larger groups sacrificing the functionality of individual robots to keep costs low. Creating low-cost robots with more capabilities is one of the key challenges in collective robotics. In this work, we developed modular, soft robots, demonstrated in The capability of a modular robot collective is dependent on the number of units within it.12 The Kilobot is the lowest cost (∼$15) robot currently available. It uses an actuation mechanism based on cheap vibrational motors. It also uses a single infared radiation (IR) sensor for both proximity sensing and communication. Through pulsing its IR transmitter, a Kilobot is able to communicate with others and by measuring the intensity of incoming IR communications. It is also capable of detecting how close other Kilobots are to it. This use of a single piece of hardware for multiple functionalities reduces the cost of robots.The use of multifunctional hardware is demonstrated by the Kilobot.13 This architecture allows the user to define the required abilities of each robot and keeps the cost-to-volume ratio balanced with respect to the task at hand. The base of the robot can then be repurposed for different tasks.Building a robot with an extensible architecture allows extra sensors to be easily added to the robot.14 uses a Hall-effect sensor for both odometry and navigation.15 This swarm robot is made of a single printed circuit board (PCB) with electromagnetic coils and uses an air table with embedded permanent magnets for low-cost locomotion. Absent magnets on the air table create magnetic landmarks that can be detected by the HoverBots. This dual use of a Hall-effect sensor allows the number of required sensors and the cost of each HoverBot to be reduced.The HoverBot16Our Linbot uses a voice coil system for actuation, audio output, sensing, and communication. This voice coil system is based on the Wormbot.17Voice coil systems are electromagnetic systems that are based on the same fundamental principles as loudspeakers; their method of actuation is detailed in 16 The voice coil systems are multifunctional components that demonstrate different behaviors over a range of different control signal frequencies. In addition to actuation capabilities at low frequencies, the voice coils can function as loudspeakers at audible sound frequencies and can be used for communications at higher frequencies.16 This communication is based on inductive data transmission.18Voice coils have been shown to be a useful actuation system for soft robotics.19We also utilize the permanent magnets in our voice coil system for sensing. The soft body of the robot deforms when it is actuated or when it is subjected to external force and we measure this deformation by monitoring a three-axis Hall-effect sensor, which responds to the relative position of the permanent magnets that are embedded on the top of the robot. This approach allows us to perform both proprioceptive sensing and tactile sensing.20 Many of these simple pairs of myocytes and nerves, placed at different positions and orientations, can produce complex actions such as prehensile movement or skeletal locomotion. Stacking units in this way can produce useful behavior for robotic applications.21–24 In this article, we provide two examples of stacking: first, peristaltic locomotion, which has been a major interest for soft robotic researchers.25–28 Second, we rearrange the units used for peristaltic locomotion into a grid to create a peristaltic table that is capable of moving objects over the surface.30 Two-dimensional (2D) matrices of actuators have been created by Kim et al.,9 Stanley et al.,10 and Follmer et al.,11 and these arrays could be adapted to allow a system designer to make a peristaltic table.The simple linear action of voice coil systems is similar to the action of myocytes (muscle cells). Myocytes can only perform a simple action, contraction, and respond to simple inputs, signals from a nerve.In this article, we have combined both computation and sensing into actuators that we arrange in a 2D array to create a fully distributed and modular peristaltic table.To allow voice coils to function similarly to nerve/myocyte pairs, they need onboard computation to control their behavior. By including this functionality, we have created linear modular robots that can be stacked together. We have developed low-cost robots—Linbots—that can be configured to produce different forms of useful behavior. Our Linbots are capable of communication, actuation, sensing, and proprioception all through their central voice coil system.The Linbot can be extended or contracted axially from its rest position, depending on the polarity of current applied to the electromagnetic coil in the voice coil system. A sketch of the actuation mechanism is shown in 31 The electromagnetic coils consist of two 12-turn coils used for the transmission circuit, and a larger 200-turn coil used for the actuation and receiver circuit. A circuit diagram of the coils is shown in We made the magnet holder, reel, and spring from acetate sheets using kirigami. Kirigami involves cutting a pattern out of the sheets and folding it into the desired configuration, the 2D patterns are shown in We used permanent neodymium magnets in our voice coil system. The internal components of the voice coil system are shown in We designed a fully integrated PCB incorporating a transmission circuit, receiver circuit, microcontroller (STM8S), voltage regulator, H-bridge (DRV8837), and Hall-effect sensor (MLX90393) as shown in 32 including I2C libraries for the Hall-effect sensor and universal asynchronous receiver/transmitter (UART) for communications. The Hall-effect measurements of magnetic field are used to calculate the displacement of the top half of the Linbot from its rest state.We wrote the control software in C and used standard peripheral libraries from ST,33 for magnetic communication to take advantage of the libraries provided by ST. The UART protocol uses a high idle line, which is pulled low at the start of a message. However, with the Linbot system, if two robots have their transmitters switched in, then all communications will be blocked. Therefore, we need our robots to switch their transmitters off when they are not sending any data. This requirement can lead to an extra byte, with a 0 value, being received at the end of each transmission. To avoid the extra bytes affecting the received data, we check for and remove these erroneous bytes.We use the standard UART protocolThis section discusses the design of the experiments used to demonstrate the capabilities of a single Linbot.To evaluate the output force of the Linbots, we designed a controllable experimental setup with a scale, ruler, and clamp, shown in rx is the extension, x is the absolute length, and x0 is the rest length. x0 is measured before starting the experiment. We increase the extension in steps of 0.05 from 0.65 to 1.1; these extensions represent the full range of motion for the Linbots. Using relative length will introduce some error to the experiments as the electromagnetic force from the voice coil will depend on absolute length. The measured mass on the scale was used to calculate the force output of the Linbot at each extension. This output force is a combination of the spring force and electromagnetic force.where We repeated the experiment on a sample of five different Linbots as a screening experiment; this sample size allowed us to measure more than half of the available Linbot population . We recorded the average force from these experiments and standard deviation at each extension. One repeat of this experiment is shown in The primary function of the voice coil system is linear actuation at low frequencies, but can also be used across a wide frequency range to provide audio output or communicate with neighbors. In this experiment, we programmed the microcontroller to vary the coil frequency from 7 Hz to 13.5 kHz. We then recorded sound produced by the Linbot. The experimental setup is depicted in To test pulse-width-modulation (PWM) control of the Linbot, we used the microcontroller to provide a PWM signal to the coil. We then observed the effect of changing the duty cycle on the actuation scheme. We first sweep through duty cycles from 0% to 100% and then from 100% to 0%. Next, we step the Linbot between set duty cycles of 10%, 20%, 40%, and 80%. We recorded the experiments using an 18-mp Canon EOS 100D camera and an EF-S 18–55 mm f/3.5–5.6 IS STM lens. We ran this experiment once to demonstrate the capabilities of the Linbots and the output is shown in Z axis of the Hall-effect sensor can be used to measure force parallel to the direction of motion of a Linbot. The X and Y axes of the sensor can measure the shear force between the two halves of the Linbot. In this experiment, the top half of the Linbot is moved along the positive and negative directions of the X, Y, and Z axes. A combination of the different LEDs on the Linbot is used to display the direction of displacement. The schematic of the experiment is shown in We designed an experiment to demonstrate the tactile sensing capabilities of the Linbot by showing that the Hall-effect sensor can be used to track the movement of the magnets in three dimensions. The We designed an experiment to investigate bidirectional communication between two Linbots and to evaluate the error rate in data transmission at different communication distances. The experimental setup is shown in In this experiment, we tested the communication between two Linbots at varying distances from one another. We increased the distance between the Linbots in steps of 5 mm from 90 to 150 mm. We measured the distance from the centers of the Linbots. The experiment is divided into two parts, where at each distance, the first Linbot transmits a stream of data and the second Linbot receives the transmitted data, then the second Linbot transmits a stream of data and the first Linbot receives the transmitted data. The transmitted information consists of 50 bytes of data. We set these data to be the numbers 11–211 in incremental steps of 4. This array of numbers varies the shape of the signal.The Linbot that receives the transmitted data knows the series of bytes it should be receiving, and when it receives a byte it checks the value against its expected value. If the value is different from the expected value, the Linbot interprets the byte as a faulty byte. In the experiment, the orange light depicts a Linbot in transmission mode and a green light depicts a Linbot in receiver mode. After receiving the stream of bytes, if there is a fault in the received data, the Linbot uses a combination of its LEDs to depict the number of faulty bytes received. The Linbot flashes a blue LED once for every 10 faulty bytes and flashes an orange LED once for the remaining single faulty bytes.The number of wrong bytes received is used to calculate the error rate at each communication distance. If the Linbot does not receive any transmitted data because it is outside the communication range, the error rate is considered to be 1. Once a distance is reached where both Linbots have an error rate of 1, all larger distances for that pair are assumed to be 1.Due to the large number of Linbots, we decided to perform a screening experiment by sampling more than 50% of the population, so that we could gain a good understanding of the overall capability of the Linbots. We chose five Linbots at random, from the population of nine, and used each one for two tests at each range. To separate the effects of transmission strength from the receiving sensitivity for each Linbot, we chose to test each Linbot twice, in a different pairing. By taking into account all of the permutations we calculated that we needed 500 binary tests of communication at each range. This experiment is shown in In addition to the experiments that show the capabilities of one or two Linbots, we performed additional demonstrations to show how the Linbots can perform collective behaviors. For the first demonstration, we designed a peristaltic conveyor to show how a collective of our Linbots can use simple individual behaviors to perform a more complex task. The system is shown in The platform consists of two parts: a backboard to prevent objects from falling off the conveyor and a base to hold the Linbots. The conveyor was given a 4° roll so that conveyed objects rest against the backboard, and a 1° pitch, so that these objects are moved uphill. For this experiment, we programmed the Linbot as a finite state machine to respond to either contact by an object or receiving a message from a neighbor. On detecting communication from another Linbot, the receiving Linbot contracts. When the Linbot contracts or is compressed, it turns on its communication and transmits information to its neighboring Linbot. The Linbot then expands to push the object to the next Linbot. Multiple Linbots performing this sequence create a traveling wave along the conveyor from one side to the other.After expansion, each Linbot enters an unresponsive state for a set amount of time to avoid responding to the signal from the succeeding Linbot in the conveyor. This mechanism ensures that the wave travels only in one direction, carrying the object with it. Schematics of the mechanism are shown in To demonstrate how Linbots can be reconfigured to perform other tasks, we designed a peristaltic sorter using the Linbots. The layout of the Linbots for this sorter is shown in As the sorter configuration uses multiple, nondormant Linbots within the communication range of one another, we needed to use an addressing system to control if a received command activates a Linbot. To implement the addressing system before starting the sorting task, when the Linbot collective is turned on, the bottom left Linbot is pressed. Pressing a Linbot will cause it to propagate a message through the collective, allowing each Linbot to know how far away, on the grid, it is from the Linbot pressed. The process is repeated with the top left Linbot. Based on the grid distances given by the bottom left and top left Linbot, each Linbot will have unique coordinates. Each Linbot will also know the relative position of the other coordinates on the grid. This addressing system is demonstrated in 34When a Linbot detects a weight placed on it, it increments a counter related to the class of the weight it believes is on top of it. When the counter of a class reaches a threshold, the Linbot sends a message to its neighbors causing them to actuate. This buildup and discharge behavior is inspired by action potentials in heart muscle cells.The message transmitted by the Linbot under a weight contains the coordinates of the signaling Linbot and the direction to move the weight. The neighboring Linbots actuate based on their relative position to the instructing Linbot. This actuation leads to the ball rolling off the edge of the grid in the desired direction. The Linbots all have the same controller and can function in any role or position within the system. The setup and behavior of the sorter are depicted in We designed the Linbots to have a size and weight appropriate for ease of fabrication, manufacturing, and assembly. They each have an inner diameter of 50 mm, an outer diameter of 70 mm, and a height of 50 mm at the rest position. This size allowed us to fit all of the electronic components on one side of the PCB while also allowing large numbers of Linbots to be space efficient. Each Linbot weighs 33 g, and the battery used for each Linbot also weighs 33 g. The Linbots are designed for manufacturability and the total bill of materials is £13.64. Further information about the components and assembly can be found in The Fabrication of the Linbot section of We expected the force output to be the sum of spring force and magnetic force between the electromagnets and permanent magnets. When the coil and magnet are fully separated, the force is given by the following:F is the total force, k is the spring constant, d is the distance from rest position, μ is the permittivity of air, r0 is the initial distance between center of the coil and center of the magnets. coilq is given by the following:where N is the number of turns, I is the current through the coil, A is the cross-sectional area of the coil, and L is the length of the coil.where When the permanent magnets enter the coil, the magnetic force between them drops, reaching zero when their centers align. At the beginning, we expect a positive force trending down linearly due to the spring force dominating. Once the centers of the permanent magnets move away from the center of the coil, we expect a rapid increase that overtakes the decrease of the spring force. Once the permanent magnets are separated from the electromagnet, further behavior will be an inverse-square relationship, summed with a negative linear term as the spring force starts pulling the magnet back.We see this expected behavior in the results from our force test, shown in The rest length of the Linbots used in the experiment ranged 46.5–51.0 ± 0.5 mm. Removing this variance would require using a machine to fold the springs of each Linbot. The error seen in this experiment is acceptable as these robots are soft and have not been designed to require a high level of precision to achieve their tasks; instead, they rely on compliance to adapt to the environment. The maximum force output was 0.81 N, which suggests that the Linbots could lift objects with masses below ∼83 g. For objects near this limit, lifting would require a controller that actuates such that it does not allow the Linbot to compress below 0.85 relative length. Below this relative length, the force output drops off and the Linbot may be unable to lift as heavy an object.We expected the Linbots to actuate at a low control frequency and transition to sound output as the frequency reaches audible levels. At high frequency, we also expect to use PWM to produce forces smaller than the maximum actuation force, and therefore, partial actuation. We demonstrated that the Linbots can actuate up to high frequencies and act as a loudspeaker. The waveform for the output sound is shown in We designed the Linbots to be able to sense the displacement of their top half relative to their bottom half. We expected accurate classification of movement for positive and negative movements along three orthogonal axes. We demonstrated this ability in We expected each pair of Linbots to show a very high successful transmission rate at separation distances below their maximum transmission range, as the receiving circuits have a threshold for signal strength that is set above the background noise. The background noise level from the receiver circuit is shown by Due to variation in the construction of the coils, the transmission strength and receiving sensitivity vary in each Linbot. This variation gives each pair of Linbots a different maximum transmission range. The effect of this variation was expected to give rise to a high success region before any pairs reach their maximum range, then a distance where the average success rate is below 1 and has a high error due to some pairs having started their rapid decrease, while others have not. The average success rate should then decrease to zero as more pairs move past their maximum range.The results of the communication test between the Linbots are shown in The results of these communication experiments are used to quantify the maximum distance for reliable communications and acted as a guideline for the separation distance between Linbots in the following experiments that used collectives of robots.We expected a collective of Linbots to be able to generate a sufficiently powerful peristaltic wave to roll a ball up a gradient. Our Linbot collective achieved this task and the result can be seen in This conveyor would be suitable for any objects that slide or roll but would have trouble moving adhesive objects. The peristaltic wave used could have been faster, as the video shows the ball sometimes getting slowed by falling into the trough of the wave. A faster wave also increases the chance that the ball would fall behind the wave and stop. The speed of the wave is limited by the gradient created for the ball to roll down. This gradient is determined by the actuation range of the Linbots and the spacing between them. In this experiment, the angle of the rising slope of the peristaltic wave is ∼12°, including the opposing 3° pitch of the conveyor. Increasing the range of motion of the Linbots or reducing the distance between them would allow for a faster peristaltic wave.We designed the Linbot collectives to be able to sense the difference between two balls, one weighing 2.5 g and the other 40 g. We also expected that they would move the 2.5 g balls off the grid to the left side and the 40 g ball off the grid to the bottom side. Our collective behaved as expected. This behavior is depicted in The signal used to classify the balls is shown in The high actuation frequency of the Linbots allows them to be used for vibrational sorting. Vibrating collections of objects allow them to be sorted by firmness or density. A single Linbot with a slanted plate attached to the top can produce the vibrations needed to sort objects. The peristaltic sorter could also be adapted for a mixture of vibrational and peristaltic sorting.Our use of inductive data transmission for the Linbots means that communication is only possible between nearby Linbots. In the future, the frequency of the carrier wave used could be increased to create far-field communication between Linbots and to enable long-range communication between dispersed Linbots.Through adding a rectifier and LiPo balance circuit to the Linbot, its battery could be charged through inductive power transfer. This would involve placing the Linbot in a strong alternating magnetic field, whose power would be coupled into the main coil of the Linbot.By writing a program to allow Linbots to be programmed via UART, we could allow for wireless programming of the Linbots. This modification would reduce the time needed to program the Linbots as an antenna could be used to program them simultaneously. It would also allow new programs and controllers to be propagated through a collective by the Linbots themselves.2C sensors to Linbot PCB I2C bus pins, we can tailor the Linbots to new tasks. For example, to make an array of Linbots respond to the distance from objects, IR range sensors can be added to a Linbot PCB.By connecting IIn large Linbot collectives, each individual robot can affect the state of other Linbots with whom it does not have a direct communication link. This complication leads to difficulties in writing effective controllers. Machine learning algorithms could be applied to allow Linbots to learn the dynamics of their collective and even to have individually tailored controllers.35 We can create these pulses with periodic broadcasts from a Linbot to neighboring Linbots. We can also use the periodic actuation of a Linbot, which can detect the actuation of neighboring Linbots through tactile signals from a flexible layer on top of them.If we have a task that requires synchronization, we can design our Linbot controllers to create a global clock. Using phase rate equalization, any detectable pulses can be used to synchronize the Linbots.Further robots could be added to make the peristaltic sorter larger without needing to rewrite the controller. This larger sorter could be used to examine large-scale behaviors of Linbots.16 The peristaltic conveyor could also be inverted to produce waves that would enable the whole assembly to locomote across a surface. Reducing the weight of the Linbots by decreasing the battery size would allow for more robust locomotion. A 2D Linbot collective designed for locomotion would be capable of steering in response to sensory information.A Linbot collective could be reconfigured for locomotion, similar to the Wormbot.The Linbots could be miniaturized for use in different applications such as tactile displays or implantable devices. By converting the circuitry into one integrated circuit, the volume of a Linbot could be reduced to about a cubic centimeter. At this size, the Linbots could use the magnets and coils from the smallest commercially available solenoids, while still using off-the-shelf batteries.Making the robot even smaller would preclude the use of available batteries. This means that either a miniature battery would need to be manufactured or the robot would need to run on miniature charged capacitors and an external power source. At smaller scales, the coil would need to be changed from being a wound wire to a planar coil cut into a PCB, and the magnets would need to be fabricated rather than bought. The limit at this level would be the minimum size of integrated circuits, which means the Linbot cannot be smaller than a few millimeters in size.Modular robotic collectives are more robust than monolithic systems. Often modular systems are limited in their functionality due to cost. Soft, modular robots also often require an outside pressure or vacuum source. We present our Linbots, our combination of untethered, reconfigurable robots with soft robotics. Our Linbots demonstrate communication, actuation, tactile sensing, proprioception, and sound synthesis. The linear motion of our Linbots sets them apart from existing modular robots and allows them to be used for tasks that require peristaltic motion.We demonstrated the abilities of individual Linbots as well as their ability to communicate with one another. We used a Linbot collective to convey a ball up a slope using a peristaltic wave. We then reconfigured this Linbot collective to demonstrate sorting. The collective was able to sort two balls based on weight and transfer them to desired bins.Our use of multifunctional hardware allows the system to be low cost without sacrificing functionality. The soft nature of our Linbots allows them to conform to their surroundings. Our Linbots provide a low-cost modular platform that can be configured for different real-world tasks. In addition, our multifunctional voice coil system can be adapted to other modular or swarm robotic systems. Our work supports the move toward reconfigurable, modular robotic platforms as useful tools for both academia and industry.This study was supported by EPSRC via the Robotarium Capital Equipment and CDT Capital Equipment Grants (EP/L016834/1), and the CDT in Robotics and Autonomous Systems. M.P.N. gratefully acknowledges support from the CDT in Intelligent Sensing and Measurement (EP/L016753/1), United Kingdom. A.A.S. acknowledges support from the EPSRC ORCA Hub (EP/R026173/1).