UBE3A gene. UBE3A is located on chromosome 15q11–13 and is biallelically expressed throughout the body but only maternally expressed in the brain due to an RNA antisense transcript that silences the paternal copy. There is currently no cure for AS, but advancements in small molecule drugs and gene therapies offer a promising approach for the treatment of the disorder. Here, we review AS and how loss-of-function of the maternal UBE3A contributes to the disorder. We also discuss the strengths and limitations of current animal models of AS. Furthermore, we examine potential small molecule drug and gene therapies for the treatment of AS and associated challenges faced by the therapeutic design. Finally, gene therapy offers the opportunity for precision medicine in AS and advancements in the treatment of this disorder can serve as a foundation for other single-gene neurodevelopmental disorders.Angelman syndrome (AS) is a rare  neurodevelopmental disorder characterized by severe developmental delay and intellectual disability, impaired communication skills, and a high prevalence of seizures, sleep disturbances, ataxia, motor deficits, and microcephaly. AS is caused by loss-of-function of the maternally inherited The online version contains supplementary material available at 10.1007/s13311-021-01082-x. Angelman syndrome (AS) is a rare ~1:15,000) neurodevelopmental disorder characterized by severe developmental delay and intellectual disability, impaired communication skills, and a high prevalence of seizures, sleep disturbances, ataxia, and motor deficits :15,000 n. AS is gUBE3A gene [UBE3A is located on chromosome 15q11–13 and is biallelically expressed throughout the body but only maternally expressed in the brain due to imprinting [UBE3A-ATS [AS is caused by loss-of-function of the maternally inherited E3A gene –11. UBE3printing –14. The BE3A-ATS . SeveralBE3A-ATS –20.UBE3A. The presence of a silent, yet functional copy of UBE3A, on the paternal allele, allows for the development of therapies directed at reactivation of the paternal copy of UBE3A [UBE3A, therapies aimed at introducing a functional copy of the gene are only required the central nervous system [UBE3A is highly conserved across species, making it possible to not only model the disorder but to test various preclinical therapies for safety, efficacy, and toxicity [UBE3A, which are theorized to be more likely symptomatic treatments, in comparison to comprehensive disease modifying therapies. Here, we review UBE3A and its contribution to AS, evaluate the strengths and weaknesses of current translational rodent models and how they will help in the transition from preclinical to clinical trials, the applicability and feasibility of emerging gene and cellular therapies for AS, as well as small molecule pharmacological approaches, and challenges that face the field of precision medicine as therapies are developed.Single-gene disorders are good candidates for precision gene and cellular therapies, and AS is particularly encouraging given the unique properties of of UBE3A , 22. Mors system , 14. UBEtoxicity –27. Withtoxicity –31. In eUBE3A encodes for a 100 kDa ubiquitin-protein ligase E3A and was initially described as an association factor between p53 and the E6 oncoprotein of various human papillomavirus types where the E3 ubiquitin ligase binds to p53 and breaks it down using the ubiquitin proteolysis system [s system . The gens system , 34. UBEs system . Interess system , 37. UBEs system , 39.UBE3A expression is correlated with the regulation of various genes involved in protein catabolism, cell cycle, brain morphology, and transcriptional regulation [While the function of UBE3A as a ubiquitin ligase protein and transcriptional coactivator is evident, the exact pathogenesis resulting from loss-of-function of the maternal allele in AS remains elusive. Interestingly, many of the missense and single amino acid insertion or deletion mutations seen in AS individuals affect the C-terminal catalytic domain, thus inferring primary ligase function, as central to mechanism , 37. Thegulation , 46. Congulation .UBE3A, that are either maternally or paternally expressed, and have been linked to multiple neurodevelopmental disorders [UBE3A including MKRN3 (makorin/ring finger protein 3), NDN (Necdin), MAGEL2 (melanoma antigen gene family member like 2), and SNRPN , while the PWS-IC is unmethylated on the paternal allele allowing for expression of those same genes. The AS-IC is located upstream of the PWS-IC and has been thought to help mediate the switch from paternal to maternal imprinting during oogenesis and act as a bipartite regulatory element with the PWS-IC [UBE3A including GABRB3, GABRA5, and HERC2 are not imprinted and biallelically expressed on both the maternal and paternal allele  or sire (Ube3am+/p−). Translational phenotypes including motor impairments detected by reduced latencies to fall off a beam and accelerating rotarod, while gait analysis demonstrated variability similar to kids with AS [Ube3am−/p+ mice [Ube3am−/p+ mice, across strains [Ube3a specific, on a C57BL/6 J background strain and [Several eletions , 66. As o Gabrb3 . The mos3 [Ube3a , 68–73.  with AS –80. Inte/p+ mice . Learnin/p+ mice . When th/p+ mice –84, and /p+ mice –87. More strains . Importarain and , 89 exhirain and .Ube3a, rather it inserts a visible tag via fusion of yellow fluorescent protein (YFP) to the c-terminus of E6-AP [Ube3a from either the maternal or paternal copy. Reporter mice have become invaluable for translational neuroscience [Another unique model is a reporter mouse that does not delete or mutate of E6-AP . These Yoscience , 93.Ube3a gene region [Ube3am−/p+ animals compared to wildtype littermate controls on a pairwise discrimination touchscreen task. Lastly, deletion animals demonstrated reduced exploration to affiliative pro-social calls, suggesting lower social communication or social cognition, currently being explored further [Recently, a rat model of AS was created using a CRISPR/Cas9 system to delete the entire 90-kb e region , 78. Imp further . Genetic further –98.Multiple small molecules are in clinical or pre-clinical development for Angelman syndrome. While precision therapy approaches may offer high future potential for disease-modifying treatment of genetically defined disorders, our understanding of these valuable therapies is relatively new. Indeed, at the time of writing, only two gene therapy products are currently FDA approved . Small molecule therapies, while not being disease modifying, can have great impact on specific symptom domains, such as seizures, having a major impact on quality of life. There is a large volume of preclinical and clinical knowledge around small molecule anticonvulsant drugs which can be leveraged for seizure control in AS.A receptor agonist. It is also known as THIP  or gaboxadol. As compared to other GABAergic drugs such as benzodiazepines, which act on synaptic GABAA receptors mediating fast phasic inhibition, OV-101 acts specifically on extra-synaptic GABAA receptors [OV-101 is a small molecule extrasynaptic GABAeceptors . These receptors . There ieceptors , which receptors  and EEG eceptors . OV-101 Preclinically, OV-101 has been shown to improve tonic inhibitory deficits in slice preparations from AS mice , 103. ItBoth trofinetide (NNZ-2566) and the follow-on molecule NNZ-2591 are small molecule IGF-1 mimetics. They activate PI3K-AKTt-mTOR and Ras-MAPK-ERK pathways and have been shown to increase synapse number and synaptic plasticity , 106. SpThere are no preclinical studies published using AS models for these compounds. However, NNZ-2591 showed positive effects on memory in scopolamine challenged rats , and NNZLovastatin was the first compound to be marketed in the statin class of compounds. These are HMG-CoA reductase inhibitors, with broad lipid-lowering effects, commonly prescribed for hypercholesterolemia. Lovastatin also acts to reduce RAS/ERK signaling , 119 andIn a preclinical study, lovastatin protected AS mice from audiogenic seizures, and reduced long burst firing in slices from these mice . This suMinocycline is a tetracycline antibiotic which can be used to treat both gram-positive and gram-negative bacteria. Tetracycline antibiotics work by inhibiting protein synthesis , so coulClinically, Grieco et al.  showed sm−/p+ mice as a result of loss of ubiquitin-mediated degradation of PTPA, a PP2A activator [m−/p+ mice, and genetically reducing expression of PTPA corrected spine morphology deficits. Pharmacologically, the authors showed that directly inhibiting PP2A using tool small molecule LB-100 was able to reduce PP2A activity, rescue reduced mEPSC frequencies, and improve motor behavior on both the wire suspension and rotarod tasks. While LB-100 is a tool compound and not a therapeutic, PP2A inhibition as an approach is chemically tractable, and represents an interesting new pharmacological target for AS. However, given its ubiquitous tissue expression, and involvement in many cellular processes, it remains to be seen if this approach can be tailored to specifically benefit AS patients.LB-100 is a protein phosphatase 2A (PP2A) inhibitor. Recently, PP2A was shown to be overactive in Ube3activator . The autSun et al. used a UBE3A-KO engineered human embryonic stem cell (hESC) model to demonstrate a biphasic effect on excitability, with KO neurons initially showing decreased spike frequency compared to wildtype neurons at low levels of current injection, but showing increased spike frequency at higher current levels , 136. Th, where the therapy is administered directly into the patient’s body. Precision therapy offers hope for many single-gene diseases in which conventional treatment has failed. Because AS is caused by the loss of functional UBE3A, gene therapies designed to introduce a working UBE3A copy or targeting the antisense silencing the maternal copy designed to reactivate the paternal allele are the most promising  approach [Ube3a-ATS downstream of SNRPN. After nuclear hybridization of the ASO to the ATS RNA, the Ube3a-ATS is cleaved and subsequently degraded. When the ASO was administered via a unilateral intracerebroventricular injection into a paternally inherited Ube3a-YFP mouse, paternal expression was reinstated for 4 months. Additionally, exon 2 deletion Ube3am−/p+ mice treated with the ASO exhibited rescue of contextual fear conditioning deficits and excessive weight gain, albeit weight gain is not a translational phenotype in AS. To date, this is the only reported ASO used in an AS model, but with the progression of ASOs across clinical trials [Building from the topotecan results, one study aimed to reactivate the paternal allele by suppression of the approach . ASO effl trials , it is eUbe3a expression in Ube3a−/p+ mice [Ube3a-ATS were described [Snord115 and Ube3a 3′ UTR and the paper by Wolter et. al. designed to target approximately 75 regions of the Snord115 cluster, both approaches disrupt Ube3a-ATS without affecting Snrpn or Snord116 expression [Ube3a upregulation. In utero injections were performed in the Wolter study with similar transduction efficiency and increased levels of Ube3a. Both studies utilized a variety of behavioral, anatomical, and functional assessments including body weight, brain size, open field, marble burying, nest building, open field and latency to fall on the rotarod task. In both studies, functional rescue was observed on multiple behavioral domains albeit on different tasks. These two manuscripts both represent the first in vivo nuclease use of Cas9 in the brain of an AS mouse model. While more work is needed to understand the optimal age of intervention, “optimal” translational outcome measures, and further studies on the safety and toxicity of this approach, they both present promising avenues to disrupt Ube3a-ATS and drive expression of Ube3a from the paternal allele.Various other engineered DNA-binding proteins such as zinc finger-based artificial transcription factors (Zinc-ATF), transcription activator-like effector-based artificial transcription factors (TALEs), and clustered regularly interspaced short palindromic repeat (CRISPR/Cas9) systems have been considered or studied as a tool to reactivate the paternal allele in AS . Both Zi/p+ mice , 57. Molescribed , 143. Inpression , 143. BoUBE3A into affected cells. This approach has been attempted by injecting an adeno-associated virus (AAV) carrying Ube3a into the hippocampus of Ube3a-deficient mice [Given that AS is a single-gene disorder, resulting from the loss of UBE3A activity in the brain, another practical gene therapy approach would be to introduce functioning ent mice . The treUBE3A were introduced into immunocompromised Ube3am−/p+ mice to provide functional UBE3A via cross correction [Ube3am−/p+ mice and rats [In a more recent study, modified autologous hematopoietic stem and progenitor cells with a lentiviral vector expressing rrection . This trand rats , 145–147UBE3A, minimally invasive, non-toxic, restore UBE3A to normal endogenous levels, and be effective at various developmental timepoints. One of the largest roadblocks faced in therapeutic development is delivery.While targeted gene therapies offer many advantages in the treatment of AS, there are still challenges to be discussed when utilizing these methods. The ideal gene therapy for AS would be specific to Delivery of a gene therapy to the required site is a challenge encountered universally. Given that AS is a CNS-specific disorder, delivery must be able to cross the BBB or be administered intracranially or intrathecally. Intracranial injections are invasive and generally not customary for a non-lethal disorder; therefore, intrathecal or peripheral administration are at the center in gene therapy design. Attention must be paid to the delivery vehicle as well  to reduce possible oncogenic or immune activation effects such as those seen in AAV research .UBE3A reintroduction does not overexpress the gene.Another challenge a gene therapy designed for AS is that of gene dose. Maternally derived duplications or triplications of 15q11.2-q13.3 are the most penetrant genomic rearrangements observed in individuals with ASD, accounting for up to ~3% of ASD cases, and indicated in/responsible for Dup15q syndrome –151. ProUbe3a in early embryonic development resulted in predictable AS behavioral phenotypes while either juvenile or adult loss of Ube3a had fewer, lessened behavioral deficits, suggesting the importance of Ube3a reinstatement earlier in life [Finally, the time point of a treatment is critical in a developmental disorder such as AS. Ideally, the hope is to treat as early as possible, but a therapy for all ages is essential. Previous rodent work has shown that a loss of  in life , 157. Th in life , but thi in life . This de in life , 57 and  in life .UBE3A, animal models of AS, emerging genetic, stem cell and precision medicine therapies and associated challenges faced in bench to bedside pipeline. Currently, there is no cure for AS and the treatments available are only useful in mitigating and managing a few of the symptoms throughout a patient’s lifetime. Gene and molecular precision therapies offer a promising glimpse into the future of precision-medicine and work in this field will likely pave the road toward therapeutic development for other single-gene disorders.Here, we review the Angelman Syndrome, the causal gene, Supplementary file1 (PDF 372 KB)Supplementary file2 (PDF 499 KB)Supplementary file3 (PDF 508 KB)Supplementary file4 (PDF 508 KB)Below is the link to the electronic supplementary material.


Pospiviroidae: their secondary structure can be subdivided into five structural and functional domains and “viroids have evolved by rearrangement of domains between different viroids infecting the same cell and subsequent mutations within each domain”; this article is one of the most cited in the field of viroids. Employing the pairwise alignment method used by Keese and Symons and in addition to more recent methods, we tried to reproduce the original results and extent them to further members of Pospiviroidae which were unknown in 1985. Indeed, individual members of Pospiviroidae consist of a patchwork of sequence fragments from the family but the lengths of fragments do not point to consistent points of rearrangement, which is in conflict with the original hypothesis of fixed domain borders.In 1985, Keese and Symons proposed a hypothesis on the sequence and secondary structure of viroids from the family  Pospiviroidae are transcribed by DNA-dependent RNA polymerase II (pol II) using the viroid RNA as a template in an asymmetric rolling circle mechanism in the nuclei of infected cells  and other viroids such as PSTV[d] and by certain pairwise comparisons such as CEV[d]-A and TASV[d] in which there is significant change from relatively low sequence homology in the P region to higher homology in the adjacent T1 [TL] and C domains” .ucAlN of HSVd with PSTVd and CEVd with TASVd, mentioned in the above sentence, are shown in kmer diagonals. In alignments  exhibit unusual relationships with respect to their terminal sequences. For example, TASV[d] shares 73% overall sequence homology with CEV[d]-A but the T2 [TR] domains are only 46% homologous … In contrast, TASV[d] shares less overall sequence homology with PSTV[d] (64%) but the T2 [TR] domains are highly homologous (90%). Therefore, TASV[d] appears to be a recombinant between the T2 [TR] domain of a PSTV[d]-like viroid and all but the T2 [TR] domain of a CEV[d]-like viroid” .Clearly, several viroid sequences show PSI values of individual domains largely deviating (>20%) from the PSI values of their full sequences . One of “TPMV [MPVd] shares 76% overall sequence homology with PSTV[d] but the T1 [TL] domains are less homologous (67%). In contrast, TPMV [MPVd] shares less overall sequence homology with CEV[d]-A (60%) but the T1 [TL] domains are more homologous (80%). Thus, TPMV [MPVd] appears to be a recombinant between the T1 [TL] domain of a CEV[d]-like viroid and all but the T1 [TL] domain of a PSTV[d]-like viroid” .ucAlnmafft X-INS-i  [li aligns with the TL, upper P, and upper C domain of CDVd to ASSVd and the TR and lower C domain to PBCVd  , the autof ASSVd  and noneli are shown in Pospiviroid, Apscaviroid, and Coleviroid, respectively. Another example is CBCVd, a member of genus Cocadviroid infecting Citrus as well as hop; already Puchta et al. [Pospiviroid and Hostuviroid. Indeed, the sequence of CBCVd aligns best to upper TL, P, and V domains of HSVd, to C and lower V domain of HLVd, to TR domain of CEVd, and to lower TL domain of CCCVd (Pospiviroid have two RY motifs (see alignment in The lack of match between recombination points and domain borders generally holds true. Further examples by JAa et al.  describeof CCCVd . Note thHostuviroid, might be a recombinant between different viroid genera [Pospiviroid, includes a TCR like other members of Pospiviroid but not a TCH as HSVd, the type strain of Hostuviroid.DLVd, a member of genus d genera , in thisSimilarly to the missing match of recombination points and domain borders, points of duplication in natural variants of CCCVd ,62 and CucAln, mafft, JAli, and vmatch of evolutionary relations between viroids of family Pospiviroidae support the original finding by Keese and Symons in 1985 that these viroids have evolved by the rearrangement of sequence stretches between different viroids infecting the same cell and subsequent mutations. Even recombination between viroids from the different genera of Pospiviroidae are likely. We were, however, not able to reproduce the exact borders of these recombined stretches and thus have doubts regarding the validity of the strict domain model as defined by Keese and Symons. For all ten species of Pospiviroid, we were able to improve the original domain borders to be consistent between species with respect to a mafft alignment. Nevertheless, the conserved sequence and structure motifs—especially those of Pospiviroid members—are sufficient to subdivide their rod-shaped structure into biologically functional sections.Our analysis with the software tools N


Flow parsing is a way to estimate the direction of scene-relative motion of independently moving objects during self-motion of the observer. So far, this has been tested for simple geometric shapes such as dots or bars. Whether further cues such as prior knowledge about typical directions of an object’s movement, e.g., typical human motion, are considered in the estimations is currently unclear. Here, we adjudicated between the theory that the direction of scene-relative motion of humans is estimated exclusively by flow parsing, just like for simple geometric objects, and the theory that prior knowledge about biological motion affects estimation of perceived direction of scene-relative motion of humans. We placed a human point-light walker in optic flow fields that simulated forward motion of the observer. We introduced conflicts between biological features of the walker  and the direction of scene-relative motion. We investigated whether perceived direction of scene-relative motion was biased towards biological features and compared the results to perceived direction of scene-relative motion of scrambled walkers and dot clouds. We found that for humans the perceived direction of scene-relative motion was biased towards biological features. Additionally, we found larger flow parsing gain for humans compared to the other walker types. This indicates that flow parsing is not the only visual mechanism relevant for estimating the direction of scene-relative motion of independently moving objects during self-motion: observers also rely on prior knowledge about typical object motion, such as typical facing and articulation of humans. Extracting independent object motion in a scene during self-motion is a challenge for the visual system: any motion on the retina might be due to either the self-motion of the observer, the motion of objects in the scene, or some combination of both sources of motion Wallach, . The theTo date, many studies have demonstrated flow parsing for extracting the motion of inanimate, abstract objects during self-motion. Studies either presented optic flow fields that simulated motion of an observer  or object information  when estimating direction of motion of other humans during self-motion. We placed different types of point-light walkers , recruited from the University of Muenster, volunteered. Participation was compensated with course credit. They were informed that they would participate in a study investigating optic flow and the motion of other beings, but were naive with respect to specific hypotheses. They gave written informed consent prior to participating. Ethics were approved by the local ethics committee of the Department for Psychology and Sports Sciences of the University of Muenster. Data of one participant were excluded from the analyses due to potential misunderstanding of task instructions.We conducted a power analysis using G*Power . Experiments were presented with an Apple Mac Book Pro with an Intel Iris Pro built-in graphics card running Matlab and the Psychtoolbox  in height and 248 cm  in width.Stimuli were scenes displaying a walker, a ground plane covered by random dots, and a fixation cross Fig. a. The sc∘ × 2.9∘ visual angle) which remained visible throughout each trial. The ground plane was defined by 350 white dots  randomly distributed across the plane. On half of the trials, forward motion of the observer was simulated by moving the camera across the ground plane along the z-axis at 2 m/s. Three walker types were used: Intact, Scrambled, and Cloud. The Intact walker consisted of 12 white dots  that were recorded from a male human walking at 1.2 m/s  and 15.9∘ visual angle in height. The Intact walker faced to the right-hand side of the screen in all conditions, thus the observers saw the profile of the walker. Five scrambled versions of the Intact walker were created (Scrambled). The Scrambled walkers consisted of the same number of dots. Scrambling was generated by spatially randomizing the x- and the y-coordinates of the dots defining the PLW but keeping the z-coordinate and the articulation parameters intact. Scrambling of the x- and y-coordinates was constrained to the maximal expansion of the intact PLW during articulation. Depending on the articulation phase, the scrambled PLWs maximally expanded 10.3∘ visual angle in width  and 15.4∘ visual angle in height. The Cloud walkers consisted of the same number of dots but in fixed random locations without any independent dot motion. They were created by removing the articulation parameters from the Scrambled walkers but keeping the x-, y- and z-coordinates of the dots. Each of the five Scrambled and each of the five Cloud walkers were presented in a pseudo-random order ensuring that each walker was presented equally often in the experiment.At the center of the screen, there was a white fixation cross . Five directions of scene-relative motion were selected for the experiment. Two directions were towards the observer (26.6∘ and 45∘), one was to the right of the screen (90∘) and two were away from the observer (135∘ and 153.4∘). Except for the 90∘ condition, there was a conflict between the biological features of the Intact walker (facing direction and articulation) and the direction of scene-relative motion because the biological features of the Intact walker always faced and articulated to the right (90∘) irrespective of the direction of scene-relative motion  to the direction of scene-relative motion of the walker. Thus, in this condition the relative size of the walker remained constant throughout a trial  and 180∘ (pointing away from the observer). Participants adjusted the line by using the left and right arrow keys of a standard Apple keyboard.Participants reported the perceived direction of scene-relative motion of the walker on the ground plane. They did so by adjusting a red line with a white tip that appeared on the ground plane at the end of the trial Fig. b. The re∘, 45∘, 90∘, 135∘, 153.4∘ for Stationary and Approach; 71.6∘, 123.7∘, 149∘, 159.4∘, 164.7∘ for No approach) design with repeated measures on all factors resulting in 60 conditions.The experiment was set up as a 2  × 2  × 3  × 5 , Approach and No approach trials were physically identical. We therefore collapsed across Approach and No approach trials when the observer was stationary for all analyses.∘, 45∘, 90∘, 135∘, 153.4∘ for Stationary and Approach; 71.6∘, 123.7∘, 149∘, 159.4∘, 164.7∘ for No approach) repeated measures ANOVAs. We accepted main effects and interactions as significant if p<.05. When appropriate, we applied Benjamini-Hochberg correction for multiple comparisons with a false discovery rate of 0.25.We tested the perceived directions of scene-relative motion using repeated measures ANOVAs. We analyzed the Stationary, the Approach and the No approach condition separately. Also, we separately analyzed the comparisons between the Intact and the Cloud walker and between the Intact and the Scrambled walker. Thus, our data analyses were set up as 2  × 5  × 2  repeated measures ANOVAs.∘ condition, i.e., the condition with no conflict between biological features and direction of scene-relative motion for the Intact walker. In this condition the direction of scene-relative motion of the observer is orthogonal to that of the walkers. Thus, the proportion to which an observer compensates for their own self-motion directly relates to the angle of the perceived direction of scene-relative motion of the walker. We defined flow parsing gain as the proportion of self-motion in z-direction that participants subtracted from the perceived translation of the walker  = 9.77, p = .001, Greenhouse-Geisser corrected, F = 398.96, p<.001, Greenhouse-Geisser corrected, F = 3.83, p = .08, ns). For the Approach condition  = 3.68, p = .01, F = 62.61, p<.001, Greenhouse-Geisser corrected, F = 0.51, p = .49, ns). For the No approach condition  = 5.52, p = .01, Greenhouse-Geisser corrected, F = 103.55, p<.001, F = 12.23, p = .01, For Stationary observers Fig. a, we fouF2.23, 2.48 = 9.7F = 7.92, p = .02, t-test, Benjamini-Hochberg corrected; No approach: t(11) = 3.50, p = .01; Approach: t(11) = 0.71, p = .49). This means that Relative size had no influence on perceived direction of scene-relative motion when it was a valid cue (Approach condition). When Relative size was consistent across all directions of scene-relative motion (No approach) perceived directions of scene-relative motion were smaller  for the Intact walker. No such bias was present for the Cloud walker. There was a main effects of Walker  = 5.03, p = .05, F = 197.43, p<.001, ∘ to 164.7∘) than in the Approach condition (26.6∘ to 153.4∘).The Relative size × Walker repeated measures ANOVA revealed an interaction between Relative size and Walker  = 342.52, p<.001, F = 1.13, p = .36, ns) and the main effect of Walker  = 1.06, p = .33, ns) were not significant. For the Approach condition  = 6.47, p<.001, F = 80.55, p<.001, Greenhouse-Geisser corrected, F = 0.17, p = .69, ns). For the No approach condition  = 2.89, p = .03, F = 84.62, p<.001, Greenhouse-Geisser corrected, F = 8.60, p = .01, For Stationary observers Fig. a, we fouF = 272.42, p<.001, F = 2.78, p = .12, ns). There was no main effect of Walker  = 2.66, p = .13, ns).The Relative size × Walker repeated measures ANOVA revealed a main effect of Relative size. This main effect was expected because larger angles were presented in the No approach condition than in the Approach condition (∘ (Approach) condition as there was no conflict for the Intact walker between biological features and direction of scene-relative motion. Paired-samples t-tests revealed that the flow parsing gain for the Intact walker was larger than for the Scrambled (t(11) = 2.59, p = .03, Benjamini-Hochberg corrected) and the Cloud walker (t(11) = 2.32, p = .04, Benjamini-Hochberg corrected). The flow parsing gains are displayed in Fig. Flow parsing gains were based on the 90In the present study, we adjudicated between two theories that describe how direction of scene-relative motion of other humans during self-motion is estimated. The first theory postulates that direction of scene-relative motion is based on self-motion information . The second theory postulates that direction of scene-relative motion of humans is based on object features . Presenting human (Intact condition) and non-human (Scrambled and Cloud condition) PLWs in optic flow fields revealed that during simulated self-motion, the perceived direction of scene-relative motion was biased towards the biological features for the Intact walker. This effect was found even when relative size of the Intact walker was controlled for (No approach condition) and could not be explained by disturbances of the optic flow field caused by the nonrigid motion of the Intact walker. Importantly, flow parsing gain  was larger for the Intact than the Cloud and the Scrambled walkers.From a theoretical perspective, differences between walker types are inconsistent with a pure flow parsing concept as found for geometric objects  which could lead to stronger biases towards biological features than for conditions with less salient conflict.When comparing flow parsing of the Intact walker to the non-human walkers we expected an overestimation for directions < 90∘).For Stationary observers, perceived direction of scene-relative motion was biased towards the biological features for the Intact walker when compared to the Cloud walkers. No such effect was found for the comparison between the Intact and the Scrambled walkers. The only difference between the two comparisons was the absence of nonrigid motion in the Cloud walkers. Previously, it was found that information about walking direction can even be extracted from scrambled versions of PLWs  affected perceived direction of scene-relative motion. This finding is in line with modulation of flow parsing when multisensory object cues were available such as spatially congruent sounds  and object information  when estimating direction of motion of other humans during self-motion.


Aspergillus flavus). This work aims to investigate the interaction between these two organisms when thriving in maize flour. It was observed that when both organisms were together, the mycotoxins detected in maize flour were far higher than when the fungi were on their own, suggesting that the presence of insects may contribute positively to fungi development and mycotoxin production. The insects in contact with the fungi were almost all dead at the end of the trials, suggesting a negative effect of the fungi growth on the insects. Both organisms interacted when in contact. This is the first study on this issue, although further investigation would benefit from clarification on the mechanisms leading to the nature of the detected interactions.It is important to hold cereals in storage conditions that exclude insect pests such as the red flour beetle and fungi, especially mycotoxin-producing ones  revealed a significantly higher concentration in the flour inoculated with both organisms (18.8 µg/kg), being lower when exposed only to A. flavus, suggesting that the presence of insects may trigger fungal development and enhance mycotoxin production. The ability of these organisms to thrive under the same conditions and the chemical compounds they release makes the interaction between them a subject of great importance to maintain the safety of stored maize. This is the first work evaluating the interaction between T. castaneum and A. flavus mycotoxin production. Stored products are greatly affected by both abiotic factors such as temperature and humidity and biotic factors such as insects and fungi, which may contribute to their degradation and a loss of quality and quantity. Among the biotic factors, fungi and insects are major threats and both types of organisms may either cooperate or compete to colonize the stored products, as a few authors have postulated . The prAspergillus flavus Link are mycotoxigenic and produce aflatoxins (AFs) that can contaminate foodstuff and become harmful to animals and humans mainly through ingestion [Several strains of the fungus ngestion ,5,6,7,8.ngestion .Tribolium castaneum (Herbst) is considered to be one of the most important key pests of stored milled grain [ed grain . It is aed grain . The adued grain ,13,14. Ted grain ,16,17. Ted grain ,22,23,24The interaction between insects and mycotoxigenic fungi may trigger fungi growth and mycotoxin production. Insects have been reported to be putative vectors of mycotoxigenic fungi in stored product conditions ,26,27. AA. flavus, which are well adapted to warm weather conditions and when exposed to higher temperatures  may stimulate mycotoxin production [T. castaneum, who may increase their feeding rate or reproduction rate, which is concerning as they are vectors of mycotoxigenic fungi [T. castaneum has also demonstrated adaptive thermal plasticity [Future climate change scenarios, predicting increases in temperature in several areas, indicate that food safety issues may be raised regarding not only mycotoxigenic fungi such as oduction ,32,33, bic fungi . In addiasticity .A. flavus strain as a contribution to a better understanding of this complex interaction and to the food safety of stored products.This work aimed to evaluate the possible influence of the insect presence in maize flour on the production of aflatoxins by a mycotoxigenic w) were estimated for different interaction assays and included three replicates for each determination (n = 3).Maize collected directly from fields was stored at −4 °C and then ground and sieved to obtain maize flour. The initial maize flour moisture content and water activity were determined using adequate equipment  and Hygrolab C1 ). The average values of the moisture content (%) and water activity (ATribolium castaneum (Herbst) , was obtained from natural populations with less than five years of rearing at the Entomology Laboratory of the Departamento de Ciências e Engenharia de Biossistemas (DCEB) of the Instituto Superior de Agronomia (ISA), University of Lisbon. The cultures were maintained at 26 °C and 65 to 70% relative humidity (RH) in a mixture of wheat flour and baker’s yeast (Saccharomyces cerevisiae Hansen) in a 95:5 w/w proportion, according to [The red flour beetle, rding to . Mass rerding to . The insAspergillus flavus Link  obtained from the Minho University Mycotheca (MUM-UMinho) was selected and maintained at 4 °C in the collection of the Laboratory of Mycology of DCEB, ISA, University of Lisbon.A mycotoxigenic strain of A. flavus-containing potato dextrose agar (PDA) plates grown for eight days by rubbing the sporulating surface with a bent needle. After filtering through a 60 μm mesh sieve to remove debris, sterile distilled water was added to the suspension to reach a concentration of 107 conidia/mL [Suspensions of conidia were prepared from nidia/mL ,39, baseT. castaneum adults; (3) fungus, with maize flour inoculated with 0.5 mL of the fungal conidia suspension (see n = 10).For the interaction assays, 40 g of maize flour was placed in each one of 40 glass flasks  and autoclaved at 121 °C for 15 min to eliminate potential fungal and insect contaminations. The experimental procedure for the interaction study included different types of assays: (1) control, with maize flour only; (2) insect, with maize flour inoculated with 80 sion see  and (4) n = 2). For the mixed assay (insects and fungi), a third sample was added due to the heterogeneous aspect of the maize flour.The interaction assays were maintained in a climatic chamber at 30 °C ± 2 °C and 70% ± 5% RH for eight weeks. After this period, the living insects were counted and the level of fungal development was evaluated by a direct observation. The maize flour samples were analyzed at the laboratories of the Food and Nutrition Department, National Health Institute Dr. Ricardo Jorge (INSA), Lisbon, Portugal, for the presence of aflatoxins according to the method described in EN15851  with a fp < 0.05) among them. Assumptions were tested with the Bartlett test for homoscedasticity and the Shapiro–Wilk test for the normal distribution of residues. After this, the data obtained were submitted to an analysis of variance (ANOVA). If the result was considered to be significant (p < 0.05), Tukey’s honest significant difference test (HSD) was performed. These analyses were completed with RStudio [The aflatoxin content of the different types of assays  was compared for detecting significant differences ( RStudio  and R-3.A. flavus conidia (fungus assay), inoculated with T. castaneum adults (insect assay) or inoculated with both organisms (mixed assay). The mixed assay increased the mortality of the adult insects. The insect assay reared only on maize flour achieved a mortality rate of 50.0% ± 52.7% regarding the initial number of adults; in these assays, other developmental life stages were also detected and counted when present with an average number (± standard deviation) of larvae (41.4 ± 45.7), pupae (2.8 ± 4.9) and adults (123.8 ± 140.2). The mixed assay (with insects and fungi) attained a mortality rate of 99.9% ± 0.4%; it was not possible to quantify further developmental stages although it was observed that the insect adults were dead before the next generation could achieve the adult stage.The maize flour was incubated under appropriate conditions of 30 °C ± 2 °C and 70% ± 5% RH for eight weeks on its own (control), inoculated with The fungal assay showed a clearly visible fungal growth. However, a more intense fungal growth was observed in the mixed assay with the development of caking not observed in the other assays .The assays conducted with insects or with fungi scored higher moisture contents and water availabilities compared with the controls although, visually, it was possible to verify that the mixed assay seemed to have an even higher moisture content and water availability . Unfortup < 0.001) with the mixed assay showing a significantly higher content of aflatoxins (AFB1) than the fungal assay (p < 0.001) as well as the control and insect assays (p < 0.001 for both); the fungal assay was also considered significantly different from the control and insect assays (p = 0.037 for both) (p = 0.144).Regarding the mycotoxin analyses, the results obtained revealed the presence of aflatoxins, mainly AFB1, in the fungal (4.3 µg/kg) and mixed assay (18.8 µg/kg) followed by AFB2 although in lower concentrations in both the fungal (0.3 µg/kg) and mixed assay (0.8 µg/kg). The content of AFB1 was significantly different among the different types of assays  . The AFBAflatoxin B1 (AFB1) revealed the highest concentration under the conditions of the mixed assay, exceeding the maximum legal level established for maize to be used as an ingredient in foodstuff (5 µg/kg ). AFB2 wA. flavus in maize flour. The insect mortality in the interaction assays  denoted a possible negative effect of A. flavus on the adults of T. castaneum as very high insect mortality was scored only when both organisms were present.The results revealed a possible interspecific competition between the red flour beetle and the mycotoxigenic fungus A. flavus growth, highlights the importance of assessing the potential health risks ahead for consumers and economic losses related to stored products. The increase in insect activity and fungi vectoring, together with an augmented mycotoxigenic production when both organisms are exposed to the higher temperatures recognized in future climate change scenarios, will also cause the spread of this problem to new geographical areas [The fact that climate change is occurring and that aflatoxins are considered a potential health threat for cereals in Europe due to temperature and humidity increases ,44,45,46al areas ,34,47.Ahasverus advena Waltl, a mycetophagous insect that may also attack the food products where fungi are growing, although this insect has more tolerance to aflatoxins [A. flavus [Aflatoxin B1, the most abundant aflatoxin detected in this study, has been shown to have a negative influence on the development and fecundity of latoxins . The siglatoxins ,29,30,49. flavus ,27.T. castaneum excretes defensive secretions, benzoquinones, which compete with other organisms; namely, fungi. Tribolium castaneum reaches its maximum benzoquinone excretion value about 40 days after the emergence of adults [A. flavus to produce mycotoxins (and/or other secondary defensive metabolites). However, this would need further investigation efforts as mycotoxin production is affected by several abiotic and biotic factors [It is important to stress that f adults . Under o factors ,9. The m factors  within t factors  when theT. castaneum is resistant to the entomopathogenic fungi Beauveria bassiana  Vuillemin, reducing fungal germination and growth due to benzoquinone-defensive secretions [T. castaneum has an efficient detoxification system and produces benzoquinones, which are indicated to be one of the reasons for this insect’s resistance to entomopathogenic fungi [T. castaneum to aflatoxins could be an important contribution to the field of novel mycotoxin control methods in the food industry, which may include the use of enzymes that promote the enzymatic degradation of mycotoxins, and new enzymes are needed [T. castaneum and A. flavus mycotoxin production.It is important to mention that cretions ,51,52,53cretions . Mycotoxcretions  althoughic fungi ,22,23,24e needed . This wa


Results showed 69% (81/118 cases) were seropositive for Q fever , brucellosis , and chlamydiosis , respectively; 82% of herds (40/49 herds) were infected with at least one of those diseases. Moreover, 40% of infected herds (16/40) had coinfection among the three of those diseases. Approximately 60% (20/33) of buck circulation showed seropositivity to at least one of the diseases, and 85% of infected bucks were seropositive for Q fever (17/20). Buck circulation between herds is a risk factor for diseases on farms (p=0.001); odds ratio  = 6.61–1,807.38). Moreover, the annual brucellosis test is a protective factor against reproductive failure cases on farms . Reproductive disorder cases can be caused by sexual transmission, so buck circulation can yield Q fever, brucellosis, and chlamydiosis in communities. This investigation is the first report of chlamydiosis infection in our area. Concerning Q fever, chlamydiosis, and brucellosis are zoonotic diseases that impact animal health and production losses. Control and prevention measures related to risk factors together with active surveillance programs should be incorporated into client education.The objective of this study was to identify risk factors related to reproductive disorders caused by bacterial infections in goats in northeastern Thailand. Two hundred twenty farms were investigated, and 49 herds were found to have clinical reproductive disorders. Moreover, 96% (47/49) of herds showing clinical reproductive failure preferred to circulate bucks between herds. A total of 118 sera, including 85 clinical reproductive disorder cases such as abortion ( Bacterial infection-induced reproductive disorders, such as brucellosis, Q fever, enzootic abortion (chlamydiosis), caseous lymphadenitis (CLA), listeriosis, campylobacteriosis, leptospirosis, salmonellosis can be found in Caprinae .Coxiella burnetii. As for clinical signs, does and nannies show abortions in the first trisemester, stillbirths, a retained placenta, endometritis, infertility, weak kids, and repeat breeding [Chlamydophila abortus. Infected doe goats show signs of reproductive failure, including abortion, miscarriage, stillbirths, and weak kids. In addition, orchitis, epididymis, and seminal vesiculitis can occur in bucks. Caprine brucellosis is caused by Gram-negative coccobacilli bacteria named Brucella melitensis. Regarding clinical signs, pregnant goats show placentitis and abortion in the second to third semesters or premature births, weak kids, a retained placenta, mastitis, metritis, repeat breeding, or infertility. Infected bucks may show signs of orchitis or epididymitis [Q fever is caused by Gram-negative bacteria called breeding . Chlamydidymitis , 3. OtheInfected herds face economic losses due to the clinical signs of infection, such as abortion, infertility, still births, miscarriage in does, and sterile orchitis in bucks, which can reduce productivity. Moreover, zoonotic diseases can be transmitted to humans.Information on bacterial infection causing reproductive failure in caprine was limited in northeast Thailand. The aim of this study is to determine the presence of antibodies against Q fever, chlamydiosis, and brucellosis, which can cause reproductive disorders in goats. Additionally, the risk factors associated with seropositivity in these infectious diseases are necessary for prevention and control measures due to their being zoonotic diseases.A retrospective study was conducted on the Caprine Brucellosis Surveillance Program being conducted by the Academic Service of the Faculty of Veterinary Medicine, Khon Kaen University, from March 2017 to September 2020. In total, 220 smallholder goat herds from 11 provinces including Buriram (4), Chaiyaphum (97), Khon Kaen (59), Kalasin (2), Mahasarakham (13), Nongbualamphu (5), Nakhon Ratchasima (5), Roi Et (7), Sakon Nakhon (21), Srisakate (3), and Udon Thani (4) were included in services.Serum samples were collected from every goat aged over 4 months based on the criteria of the Brucellosis Surveillance Program . In totan = 70), abortion with arthritis (n = 1), orchitis (n = 3), repeat breeding (n = 6), sterility (n = 1), and weak kids (n = 4), and bucks' circulation between herds (n = 33) was included in serological testing for Q fever and chlamydiosis.Twenty-two percent (49/220) of smallholder goat herds were found to have clinical reproductive disorders in at least one case on farms in 8 provinces. A total of 118 were found to have reproductive disorders, such as abortion . The procedures were carried out according to the instructions of the manufacturer. The optical densities (OD) of the samples and controls were measured at 450 nm using a spectrophotometer. The S/P% was determined based on the OD following the equation:IgG antibodies against The results were classified according to the manufacturer's instructions as follows.IDEXX ELISA; S/P% > 40, 30 < S/P% < 40, and S/P% <30 were positive, suspect, and negative samples, respectively. The sensitivities of the test were 100% and 89–95% for the Q fever and chlamydiosis tests, respectively. The specificity of the test was 100% for both tests .One hundred eighty-nine owners were interviewed for their management of risk factors associated with the presence of disease. Factors included herd structure and herd and health management especially, history of buck in herd, movement and quarantine, personnel hygiene for feeding animals, barn cleaning, and giving birth to animals during parturition. Additionally, the herd's annual brucellosis testing was included in the questionnaires.Data were analysed using Microsoft Office Excel 2016 and MedCalc® version 19.8 . The univariate analysis of risk factors was performed to identify variables associated with the positive herds at a significance level of 0.05 as the random effect in the model.The spatial distribution was analysed using Quantum GIS. The geographic coordinate system was used to produce the map for epidemiologic analysis.n = 18/40), 25.0% (n = 10/40), and 30.0% (n = 12/40), respectively. In addition, 16 herds (40.00%) had coinfection for at least two of three diseases.Based on clinical evidence of reproductive disorders in this area, 81.63% (40/49) of the investigated herds already had bacterial infection due to seropositivity against Q fever, chlamydiosis, or brucellosis infection. Only 9 herds  did not show antibodies against these bacterial infections by serological testing. Seropositivity to Q fever, chlamydiosis, and brucellosis testing in the herd was 45.0% , 15.25% (18/118), and 6.78% (8/118), respectively. Abortion  was the most common reproductive disorder in this area. Other clinical signs, such as orchitis, repeat breeding, sterility, and weak children were found in less than 15%. Interestingly, 41 herds shared 33 circulations of bucks with other herds, and 60.6% of bucks (20/33) were seropositive with at least one of these infections. Eighty-five percent of infected bucks were seropositive for Q fever (17/20), followed by chlamydiosis in approximately 15% (3/20) and brucellosis at 5% (1/20). Seropositivity for brucellosis was the lowest in proportion in this area in comparison to Q fever and chlamydiosis. This might have been due to the active surveillance program for brucellosis, which screens for brucellosis annually in registered goat herds [Clinical reproductive disorders and bucks' circulation related to serological testing are shown in p=0.001; odds ratio (OR) = 109.29; 95% confidence interval (CI) = 6.61–1,807.38). In addition, brucellosis testing annually was a highly significant protective factor for reproductive failure cases on farms  showed that 93.93% of bucks (31/33) were seropositive to at least one Q fever, chlamydiosis, or brucellosis infection among the 41 herds.The spatial distribution of three of those diseases is shown in 2Based on this investigation, the significant factor for controlling and preventing those diseases is avoiding bucks' circulation between herds. Although the serological screening of brucellosis is the only test in the Thai national brucellosis surveillance system, annual serological brucellosis testing was found to be a significant protective factor against bacterial reproductive disorders infection in herds. Additional advice for introducing animals, especially bucks, should be screening not only for brucellosis but also for Q fever and chlamydiosis.Previous studies showed clinical cases of brucellosis and Q fever in humans related to livestock animals in Thailand –11. Q feBurkholderia pseudomallei [Corynebacterium pseudotuberculosis [Leptospira noguchii [Campylobacter infection, listeriosis, and salmonellosis; other protozoa infections, such as neosporosis and toxoplasmosis; and virus infections such as Pestivirus infection.In northeastern Thailand, reproductive disorders have been reported to be caused by other diseases, such as epididymo-orchitis caused by domallei , orchitirculosis , abortionoguchii . However


For biomedical applications, smart materials that are used as sensors or actuators have to match some criteria, especially bio-compatibility and softness. Smart polymers are candidates that fulfill these two criteria. A sensitivity to compression is created by adding magnetic particles to a compressible foam polymer. A foam-based composite is fabricated for its small Poisson’s ratio, which enables significant compression, up to 50%. This large compression induces a change in its magnetic properties, which can be detected using coils. By setting the sensing coils as a compact array of 3 × 3, the sensor successfully detected and localized an applied deformation. For some applications where sensing or actuation is needed, it is important to avoid rigid materials, such as soft actuators for biomedical applications . Artificϕv of the composite [ϕc the composite conductivity increases drastically. By compressing the composite, the electric conductivity then increases [Smart polymers can be used in the reverse function: as a sensing material. A typical soft composite sensor technology relies on its electrical conductivity (or resistivity). Electrical conductivity is a function of the filling factor omposite , definedncreases . The larBy using magneto-rheological elastomers (MREs), which can provide a good magneto-elastic coupling , an induIn this article, a more compact system is designed. A single MRE layer is used as the sensing part of the sensor. Instead of using a continuous matrix, like the usual silicone rubber of MRE, a porous foam is used as the host matrix. Porosity in materials is known to reduce the effective Young’s modulus ,10; the 2 surface, the device can detect the point of pressure application; hence, it can mimic the tactile sense of the skin. Deformation up to 50% was recorded and successfully localized. The equivalent force range detected was up to 4.5 N with a resolution of 0.29 N.The deformation/force sensor device studied in this article uses a unique and simple smart layer: bilayer composites using MRE materials used to sense applied deformation ,7. Combiϕm = mFe/mtot = 70%; mFe and mtot are the particles’ mass and the total mass, respectively. Particles were first mixed with the solution and hand-stirred for 5 min; then the curing agent was added. The mixture was molded as a membrane with a controlled thickness, a 3.1 mm thick sample. The curing time was about 20 min at room temperature. Once cured, the membrane was cut into 20 × 20 mm2. The experimental materials consisted of a polymer foam, filled with magnetic particles. These magnetic particles were spherical carbonyl-iron particles  with a diameter D50 = 6.0–7.0 µm. The composite expanded and large pores were formed with size of up to 1 mm. By image analysis, performed using ImageJ software analysis , the picd = 6 mm and with an initial inductance of 7.2 µH. The nine coils were placed into a 3 × 3 array with dimensions of 20 × 20 mm2, as seen in The composite is the element that reacts to the deformation. For the detection, an array of nine coils was used. Each coil acts as a touch-sensitive site, or taxtel . These cThe measurement of the inductance of each coil was performed at different levels of deformation. Inductance was measured using an impedance meter  in LCR mode at the frequency of 20 kHz. Uncertainty on the inductance measurement was 1 nH.The deformation was applied using a three-axis stage from COMS , and the pressure tip was 3D-printed with ABS to avoid any eddy current interference from any metallic part. The deformation area was chosen by positioning the tip over the XY plane. The deformation was obtained by pushing the tip down (Z axis) into the composite by steps of ΔZ = −100 µm.F divided by the initial specimen cross-sectional area (A0 = 3.55 mm2) as [The mechanical properties of the sample were measured with a tensile/compression tester . The sample was placed between two plates to apply the compression. Force was applied with a rate of 4 mN/s with steps of 0.2 N up to 2 N. Displacement and force were recorded during this compression test. Engineering stress was then obtained from the measured force  mm2) as ,19:(1)σ=t divided by the initial specimen thickness (t0 = 3.1 mm) as [The compressive engineering strain was obtained from the measured displacement Δ1 mm) as ,19:(2)ε=t is taken as negative for compressive tests, so the compression ε defined by Equation (2) is also negative in this article.The change of thickness ΔThe sample was tested with a vibrating sample magnetometer  for measuring the magnetization curve of ±10 kOe (~800 kA/m). Before the sample experimental measurement, a Ni sample was first measured to adjust the equipment settings to match with the theoretical magnetic moment of the Ni calibration sample. After this calibration procedure, the magnetization loop of the magnetic composite was performed. By adjusting the clamping system, a deformation could be applied to the sample for the magnetic measurement.The absorbance spectrum measured on the pure foam is presented in The composite force versus displacement was measured, and the obtained stress versus strain plot is presented in E = 200 kPa, which corresponds to soft materials such as elastomers, especially porous polymers.From this strain-stress curve, the linear approximation provided the value of Young’s modulus of The magnetization versus magnetic field is presented in Msat is directly proportional to the magnetic particles’ magnetic saturation, Msat_p = 1.72 MA/m [ϕv. The filling factor is defined as the volume represented by all the CIPs  inside the sample volume (Vtot) as:For the measurement with no deformation applied, the curve did not present any magnetic hysteresis because of the very soft magnetic properties of the CIP-CS particles. This curve also provided the information about the magnetic quantity inside the material; the magnetic saturation .72 MA/m . The proMsat = 23,500 A/m pointed to a filling factor of ϕv = 1.35%. Sample magnetization was also recorded at a different level of compression ε defined according to Equation (2).The corresponding value of µr is then obtained from the slope dM/dH around H = 0, which traditionally refers to the magnetic susceptibility χ of the material. The relationship between susceptibility and relative permeability is µr = χ + 1. The different relative permeabilities for different compression levels are plotted in The magnetization loop is affected by the compressive strain. The compression yields to an increase in the magnetic saturation. The relative magnetic permeability ϕv = 1.35%. It can be observed in ε. The change is, however, relatively small; the initial permeability was µr = 1.06 and the largest permeability was µr = 1.11 for deformation of −50%, and the increase in permeability was Δµr = 0.05. This kind of increase in permeability with compression was also observed by using a permeability meter [The relative permeabilities are slightly larger than 1, meaning that the susceptibilities are slightly larger than 0, denoting a weak magnetic behavior. This is coherent with the low filling factor of the composite of ty meter  and simity meter , is:(4)μµr  with deformation, assuming a large compressibility of the matrix, can be expressed by [Moreover, the change in permeability Δessed by :(5)Δμrϕvµr(ϕv) = 1.04 and Equation (5), assuming a compression of ε = −0.5, provides Δµr = 0.04. These values are relatively close to the values in Equation (4) provides This change in permeability with compression is the main sensitive effect in our sensing device, as will be demonstrated in the next section.The first test was the measurement of the inductance of coils as the deformation was applied on the composite above coil L1; see i(ε = 0), where i = 1–9, were slightly different for each coil; this can arise from connections but it is also important to point out that these were all within the measurement uncertainty. More important are the trends of these curves: some have a positive slope, whereas others exhibit a negative slope.Depending on the selected coil for the inductance measurement, the behaviors are different. The initial inductances L1(ε ≠ 0) > L1(ε = 0). This curve behaved as a line in a first approximation, and a linear coefficient αi can be defined as:The behavior of the L1 coil inductance with compression provides some indications for the measurement interpretation. This coil is placed just below the compression during the measurements of the curves presented in α1 was negative. In α1. The negative value of coefficient αi implies a compression.In the case of the coil inductance L1, the coefficient 1, α2, α3, and α4 have amplitudes as |α1| >|α2| ~ |α4| >|α5|. When looking at the disposition of the coils, coil L1 was placed directly under the applied compression, coils L2 and L4 were equidistant to L1, and coil L5 was located a little farther. As the deformation was applied at point A, it is natural that the deformation was the largest at point A and decreased its amplitude with an increasing distance. This means that the αi were actually mapping the local deformation amplitude.The other coil inductances that presented a negative slope are L2, L4, and L5, as seen in i coefficients was then interpolated into an α map, and this α map is shown in By constructing a map of those coefficients according to the position of their respective coils, the position of the compression applied on the composite can be detected. For instance, α map for compression above coil L1 shows two zones: a blue-green area and a yellow-red area. According to the color scale bar, the blue-green color corresponds to α < 0. The area with low α corresponded well with the position of applied deformation. A second test, where the compression point was located above L5, is presented in α map is shown in The resulting α > 0 were located far from the compression area, as seen in Finally, the coefficients µr = 1, so the coil detected a lower effective permeability in its neighborhood. This gap then increased as the deformation grew stronger, making the measured coefficient α > 0 on this area. In this presented experiment, the positive α was not necessarily because of the dilatation of the material. This issue can be solved by gluing the magnetic layer onto the coil surface. This was not performed in these experiments because other samples might be tested in future experiments.It was optically observed that the composite was mainly compressed where the compression was applied, and it was lifted at the area where it was not compressed b becauseA = πd2/4), the local force can be calculated. The inductance behavior of L1 with the deformation, extracted from Finally, the locally applied force can be measured using this system. By combining the stress-strain curve presented in The presented experimental setup could detect local deformation and the corresponding applied forces. By approximating a linear dependency and assuming a measurement error of 1 nH, a force resolution of 0.29 N could be achieved in a measurement range of 0–4.5 N.3 layer. As it experienced uniaxial compression, its magnetic permeabilities were increased, especially the permeability. This change in permeability was successfully detected by a coil array. The stress versus strain curve in compression was recorded, and Young’s modulus was measured. The applied deformation could be converted into applied force. A deformation was locally applied to the magnetic layer. The amplitude of deformation was recorded up to 50% in compression. A map of the inductance change was constructed, giving information on the surface affected by an applied deformation or force.In this article, a magnetic composite was used as an active material sensible to deformation and force. The magnetic composite consisted of a porous (45%) elastomer filled by magnetic particles (1.35%). A sample was shaped in a 3 × 20 × 20 mmThis system has shown sensibility to (1) the amplitude of the deformation (or force) and (2) to the point of application of this deformation or force. The system was capable of detecting and quantifying a local force applied.The whole system was compact, as the magnetic layer was 3 mm thick and the coil 2 mm in height, with a diameter of 6 mm. This system is very light and easy to construct. It is believed that it can be convenient for tactile sensing in robotics science.


Zinc finger proteins are transcription factors with the finger domain, which plays a significant role in gene regulation. As the largest family of transcription factors in the human genome, zinc finger (ZNF) proteins are characterized by their different DNA binding motifs, such as C2H2 and Gag knuckle. Different kinds of zinc finger motifs exhibit a wide variety of biological functions. Zinc finger proteins have been reported in various diseases, especially in several cancers. Hepatocellular carcinoma (HCC) is the third leading cause of cancer-associated death worldwide, especially in China. Most of HCC patients have suffered from hepatitis B virus (HBV) and hepatitis C virus (HCV) injection for a long time. Although the surgical operation of HCC has been extremely developed, the prognosis of HCC is still very poor, and the underlying mechanisms in HCC tumorigenesis are still not completely understood. Here, we summarize multiple functions and recent research of zinc finger proteins in HCC tumorigenesis and progression. We also discuss the significance of zinc finger proteins in HCC diagnosis and prognostic evaluation. Zinc finger proteins (ZFPs), which constitute the largest transcription factor family with finger-like DNA binding do- mains, play a significant role in multiple biological processes. ZFPs primarily function as transcription factors in tumorigenesis and tumor progression. Transcription factors (TFs) are proteins that play a vital role in complicated biological processes, such as metabolism, autophagy, apoptosis, immune responses, stemness maintenance and differentiation. TFs regulate transcription of genes by recognizing or binding to DNA sequences directly , 2. So fHepatocellular carcinoma (HCC) accounts for a major part of the global cancer category. It accounts for approximately 75–80% of primary liver cancer . AccordiThe BTB domain  is a multifaceted protein-protein interaction motif found in whole eukaryotes. At the same time, it has been reported in previous literature that some poxvirus proteins are similar to a part of ZFPs, like ZID, GagA and ZF5; this domain has been named as POZ (poxvirus and zinc finger) domain , 17. As Scan domain, a highly conservative domain, consists of 84 amino acids rich in leucine residues, is also known as the leucine-rich region , 29. To Krüppel-associated box (KRAB) domain-containing zinc finger proteins (KZFPs), which have been reported to exist only in quadrupeds. The human genome encodes approximately more than 350 KZFPS. In recent reports, KZFPs mainly inhibit transposable elements (TEs) by recruiting transcriptional regulators and heterochromatin formation and DNA methylation in embryonic stem (ES) cells , 35. TheThe SET structural domain is widely found in eukaryotes and consists of approximately 130 amino acids and is named after the Drosophila proteins Suppressor of variegation 3–9 (Su (var)3–9), Enhancer of zeste (E(z)), and Trithorax (Trx). The biological behaviours in which the SET structural domain is involved are primarily associated with the methylation of substrates. For example, many SET domain-containing proteins can mono-, di- or trimethylation of their lysine substrates using the cofactor S-adenosyl-L-methionine (SAM), these catalyzed targets include histones and many non-histone substrates. In addition, the SET domain does not usually exist as a stand-alone entity, and in many proteins, it co-exists with other protein structural domains and interacts with proteins that regulate its catalytic function , 45.Among eight different groups of zinc fingers, the most classical zinc finger is the C2H2-like finger . As the C2H2-type ZFPs generally contain added anatomic domains, such as BTB , the Krüppel-Associated Box (KRAB) domain, SET domain and SCAN  domain . These sThe domains of ZFPs are not only diverse, but the functions of these domains are also very distinct. For example, although both the BTB domain and the SCAN domain can bind to the protein, but the BTB domain mostly inhibits the transcriptional activity of the target gene by recruiting transcription co-repressors, while the SCAN domain causes oligomerization of target proteins through its specific binding , 31. In Among the numerous biological functions of ZFPs, transcription regulation occupies a highly prominent position. ZFP relies on the domain-containing zinc finger in one segment to bind to the promoter region of the target gene and act as a transcription enhancer or transcription inhibitor. In addition to transcriptional regulation, ZFPs can induce protein-protein interactions, bind to RNA, and interact with proteins and RNAs in the meantime –55.As the most prominent role of ZFPs, the functions of transcriptional regulation are mainly reflected in both transcriptional promotion and transcriptional repression. It has been recently reported that ZEB1 (Zinc finger E-box binding homeobox 1) can bind to the HDGF (hepatoma-derived growth factor) promoter to stimulate HDGF transcription in EC  cells . AnotherAs well as functioning as transcription factors, numbers of ZFPs that can induce protein interactions have also been identified in recent years . For exaSome ZFPs also have RNA binding properties, and these ZFPs are generally thought to be involved in post-transcriptional regulatory processes such as mRNA maturation splicing and degradation , 61. RecAn increasing number of studies have shown that metabolic manipulate and signaling pathways are closely linked, not only in normal cells but also in cancer cells . RecentlZFPs like Snails  and the E-box binding proteins ZEBs  can also affect the process of cell differentiation directly or indirectly . GoossenFew ZFPs have recently been reported in immune-related processes such as immune response, immune homeostasis, and cytokine production in emerged new studies , 78.Gfi1, which function as a transcriptional repressor, is a zinc finger protein that participates in diverse development contexts. Jin et al. demonstrated that Gfi1  played a limiting role in the inflammatory response induced by endotoxin in mouse lung, and its regulatory function in alveolar macrophages as downstream of LPS (Lipopolysaccharide) receptor (TLR4) and upstream of TNF further elucidated the mechanism of Gfi1’s role in the endotoxin response . By the CCCH zinc finger proteins consist of several CCCH zinc finger domains, which are composed of one histidine and three cysteines. A small number of CCCH zinc finger proteins have been reported to play important roles in immune responses , 83. ForIn the past decades, the role of ZFPs has been reported in various of cancers including nasopharynx, esophagus, lung, gastric, colorectal, breast, thyroid, prostate, ovarian cancer , 90–97. The cell cycle regulation cannot be separated from cyclins, CDKs (Cyclin-dependent kinases) and CKIs (Cyclin-dependent kinase inhibitors) . CDC6 (cThe process of apoptosis is induced by two distinct pathways: the intracellular pathway or the extracellular pathway . In the Cancer stem cells (CSCs) are responsible for self-renewal, maintenance, and the growth of tumors . The abiThe epithelial to mesenchymal transition (EMT) plays a meaningful role in the early steps of metastasis of tumor . In a prCellular metabolism functions as a flexible network not only in normal tissues but also in the development of malignancies . MetabolA20, also known as tumor necrosis factorα-induced protein 3 (TNFAIP3), is an E3 ubiquitin ligase containing ring finger domains and is also a hotpot in immunoregulation , 142. ReLiver is the main organ where immune response and immune tolerance occur, and many immune reactions in normal hepatocytes and HCC cells may impact on the immune function of the liver . RecentlIn most conditions, ZFPs tend to play only one of the two effects, anti-oncogenic or oncogenic. However, during the past few years, some studies have shown that one ZFP may play both oncogenic and tumor-suppressive roles in HCC Fig. . ZNF191,As one of the largest transcription factor families in the human genome, ZFPs play diverse roles in cell biological functions, such as cell differentiation, apoptosis, transcriptional regulation, cell metabolism, immune response. In the past decades, there have been more reports about the role of ZFP in cancer, especially in HCC. HCC is still one of the cancers with the highest mortality rate in the world with many unsolved problems for early diagnosis and postoperative treatment. In this review, we described the role and related mechanisms of ZFPs in the development of HCC.Above all, zinc finger motifs have different types, including Cys2His2 (C2H2) like, Gag knuckle, Treble clef, Zinc ribbon, Zn2/Cys6, TAZ2 domain like, Zinc binding loops and Metallothionein. The C2H2 zinc finger motif is the largest of all zinc finger motifs. In the C2H2 motif ZFP, in addition to the zinc finger structure, also contains other common domains, such as the KRAB domain, the BTB domain, SCAN domain and SET domain. These domains could bind to DNA, RNA or proteins to function. Secondly, ZFPs were also reported in regulating post-transcriptional modification and protein-protein interaction. Finally, ZFPs affect the development of HCC in different processes, such as cell cycle, apoptosis and even metabolism. Among them, ZNF191 shows a distinct effect in the early and late stages of HCC. Some ZFPs have also been reported as prognostic factors in HCC. For example, the high expression of ZBTB20 and ZNF689 in HCC patients was strongly associated with poor clinical prognosis as well as high recurrence rates , 155. InIn a word, ZFPs play an important role in tumorigenesis of HCC, and more mechanisms need to be further studied. The targeted drugs of ZFPs in HCC need to be further explored.