In addition, ApE provides a platform for creating visually appealing linear and circular plasmid maps. It is available for Mac, PC, and Linux-based platforms and can be downloaded at https://jorgensen.biology.utah.edu/wayned/ape/.A plasmid Editor (ApE) is a free, multi-platform application for visualizing, designing, and presenting biologically relevant DNA sequences. ApE provides a flexible framework for annotating a sequence manually or using a user-defined library of features. ApE can be used in designing plasmids and other constructs  DNA visualization software must 1) annotate features and depict DNA features graphically, 2) simulate molecular cloning techniques and 3) generate visually appealing output for figures.Good DNA visualization software applies meaning to a string of DNA bases. Fundamentally this requires flexible annotation—applying names to a region, and visualization of functional regions—applying pictures to show the spatial relationships between sequence regions. Every piece of the DNA should be annotated with its biologically relevant attributes. In addition, a biologist must be able to identify subsequences such as restriction enzyme recognition sequences, recombinase recognition sequences, and overlapping end sequences that are useful for particular recombinant techniques.in silico simulation of common DNA manipulations, such as restriction digests or Gibson cloning. By manipulating DNA in silico, a biologist can ensure that recombinant constructs include functionally complete pieces that have the DNA in order and in frame. In other words, good software allows a researcher to synthesize a working plan. This might be working backwards in silico from a desired product to determine the needed inputs. Conversely, it allows a researcher to start with a given set of available plasmids and work in the virtual laboratory to generate possible products. Finally, visualization software can be invaluable for determining whether an analytic result—a DNA sequence, a diagnostic PCR or restriction digest—has generated the expected product. The scientist can use the software to align sequences or simulate gels of each step to confirm their work.Good DNA software also provides powerful Finally, good DNA software can generate visually pleasing output with a flexible level of detail. This representation should be easily exported in an open and widely used text or graphic format. For example, text output can be used generate class reports, student theses, or manuscripts for publication. Similarly, graphical output can be used to generate meeting posters or slides for class reports or conference presentations.Because of this critical need for visualization software, many DNA visualization programs have been written. Many of these are written by researchers themselves to solve their own needs in the lab. Among these are Serial Cloner . Often tWe have taken the long view to solving this problem. ApE is a freely available program written over the last 17 years by a molecular biologist for molecular biologists. Thus, it leverages the insider knowledge of what makes a successful DNA editing program. Further, the long-timeframe approach has allowed the program to become both highly versatile and streamlined—ApE now rivals the commercially available packages in both its diversity of features and its visual outputs. Importantly, unlike commercial packages, its free availability makes it well-suited for use in small labs or teaching labs.ApE is written in Tcl/Tk. Current distribution of ApE is with Tcl/Tk version 8.6.11. There are ready-to-run versions of ApE for Windows, MacOS, and Linux systems.For Windows, the program is packaged into a self-contained tclkit  using thThe exe file was edited using Resource hacker  to contaFor MacOS, ApE is packaged as an application bundle. The executable files in the bundle were generated from Tcl and Tk source . The curApE can be run on Unix/Linux systems using the Tcl/Tk windowing shell interpreter, wish, which is available as source code or precompiled binaries for most *nix . The wisWe have also run ApE within the Android operating system using AndroWish  as the TThe usefulness of a program can be judged on three factors: flexibility of input, flexibility of data processing, and flexibility of output. To make ApE widely usable, we have endeavored to write procedures to read as many DNA sequence file types as possible. ApE reads FASTA or raw ASCII, GenBank , EMBL, GApE saves DNA data in a GenBank-like file format that is designed to be understood by most parsers that can parse Genbank files. This format is open and human-readable text, so saved data is not confined to a proprietary, binary format. In addition, many other programs and open-source libraries such as BioPerl or BioPython can read this format easily. Although it is based on GenBank, ApE files contain additional information not specified in the GenBank specification. First, sequence-wide information is stored as a special COMMENT line that begins with the text “ApEinfo:” Second, each feature has additional feature-specific formatting data stored in feature qualifiers that begin with “/ApEinfo_.” Some GenBank parsers require qualifiers to be part of a controlled vocabulary, so ApE has a user-specified option in the preferences window to save files without this information. Only the COMMENT fields of the GenBank header are visible and editable in the ApE interface , howeverTo make ApE as flexible as possible for processing and visualizing user data, ApE stores several data files as human-readable text files. This allows users to store multiple versions of the files for different purposes, or trade useful variants with others. ApE uses this modular framework for the restriction enzyme set, the feature library, gel ladders, graphical arrowheads and user preferences. The restriction enzyme files store recognition and cut sites, methylation specificities, and user specified enzyme “groups,” which can be used in limiting enzyme searches (see below). Included with the distribution is a basic default set of enzymes, as well as several other enzyme database files, such as a set of all commercially available enzymes. DNA ladders for use in virtual agarose gels are stored in a file that can be edited using a ladder editor dialog within ApE. Arrowheads files are available to the user to customize the graphic map window. The “ApE Defaults.txt” file stores over 100 default values for many user-specified parameters between sessions.C. elegans, mouse, yeast and generic plasmid features, but there is a built-in system for adding new libraries or editing the default libraries.Finally, ApE includes a folder of feature definition library files. Feature definitions are designed to provide a rich and flexible matching paradigm. Definitions include all of the characters of the IUPAC degenerate nucleotide code, with all sequence bases required to be within the degenerate set at each position for a match to be noted. There are two variable length wild-card characters # and +, which match any continuous string of nucleotides. Definitions can contain < and >; any characters before < and after > are not required to match in the search stage, but after a match is found sequences continuous with the match that also match the pre- or post-sequence are included in the final match. Finally, the definitions can contain either uppercase or lowercase characters. Once a match has been found, uppercase characters are noted as part of the feature, while lowercase characters are gaps in the feature. This allows for feature gaps such as introns, as well as searches for specific bases within a given context, for example common or important SNPs. If a definition has only lowercase characters, all of the characters are included in the feature. Currently, ApE ships with default feature libraries for Many of the procedures within ApE could be used as stand-alone, command-line functions or incorporated into other DNA analysis projects. ApE has several basic analysis functions such as reverse complement, complement, translate, reverse-translate, search with IUPAC degeneracy codes, search for amino acid sequences in a translated DNA sequence, and melting temperature calculation. ApE also implements the DNA Strider algorithm for fast hexamer searching for restriction enzyme patterns , which iApE implements pairwise alignment of two DNA sequences using a Needleman-Wunsch (NW) alignment algorithm with an affine gap penalty. Because this algorithm is processor intensive, the alignment algorithm first uses a simple heuristic algorithm for doing a first-pass, block-based search for locally identical sequence matches, which are then used as boundaries for aligning non-identical blocks by the NW algorithm. If the sequences have no major matching regions, the user can further specify a maximum value for mismatched regions to be aligned by the NW alignment algorithm. If a region between matching blocks has a product of lengths of each mismatched sequence region, the region is not aligned, and will be highlighted in black text in the resulting display. Once a pairwise alignment is made between the reference and each comparison sequence, the alignments are combined into a single alignment by adding gaps to each sequence; no attempt is made at multiple sequence alignment.ApE has many ways to output and share data. For text-based visualizations or analysis windows, ApE can save an output file as plain text, or as formatted rich text format (RTF) files, which preserves color background highlighting and other text formatting. On Mac OS, formatted text can also be copied to the clipboard in RTF format. For graphic visualizations of data, for example, graphic maps or virtual agarose gels, ApE can save the data in four formats: encapsulated postscript (eps), scalable vector graphics (svg), OpenXML-based Power Point (pptx) and portable document format (pdf). An additional format, Windows Metafile (wmf), is available on Windows systems. All of these formats retain the information in vector format, so that they can be edited when opened in a vector editing program, such as Inkscape or Adobe Illustrator. For users who use LabArchives to store their laboratory notebook, ApE has a direct interface to the LabArchives internet portal, so that analysis windows can be directly uploaded to a user’s account. For making presentations, pptx files can be read into Power Point, Keynote or Google Slides. Finally, on Mac and Windows, ApE is able to directly output windows to an attached printer with formatting preserved. For DNA Sanger sequencing files, the data are scaled to fit within the printed page, with a user-specified number of lines per page. This wide variety of output formats and modalities should make ApE useful for saving an analysis in a laboratory notebook, for presenting the analysis on slides, for archiving the analysis in a database, or sharing the analysis on the internet.https://www.youtube.com/playlist?list=PLXd4WouGm92muGl4mJx5EvUFVSWZKdyuA.ApE has many functions for working with DNA. First, sequences can be annotated, applying names to regions of a sequence. This is described below under “Construct Features.” Second, it can edit DNA, described below under “Basic Editing.” Third, it can generate formatted text or vector graphic representations of the sequence, described in “Sequence Visualization.” Fourth, as described in “Restriction Site Analysis,” it can locate enzyme recognition sites in a sequence and simulate agarose gels of restriction digests. Fifth, are the “Molecular Techniques Simulators,” including simulators for a Restriction ligation reaction, a Golden Gate reaction, a Golden Gate reaction designer, a Gibson Assembly reaction, a Recombinase/Integrase mediated joining reaction, and PCR reactions. Finally, ApE provides several “Analysis Tools,” including alignment of Sanger sequencing to a reference sequence, a dCAPS genotyping designer, direct input into the NCBI BLAST server, and several other minor tools. A video tutorial series describing many of the features of ApE is available at YouTube: via an automated library search. Features can be visualized in four ways: as text in a table at the top of a sequence, as a text appearing when pointing to a sequence, as a graphical representation when pointing to a sequence line, or as a small graphical summary at the right side of the sequence window.A key role of Ape is to locate and highlight functionally important sequences, called “features.” Features can be added to a sequence manually or In the main sequence window, features are indicated as highlighted text . In addiAn important aspect of ApE is that features can be added to a file by using a predefined or user-defined feature library to scan the entire sequence. Feature libraries consist of lines of text referred to here as feature definitions. Each feature definition includes a name, a sequence of the feature , and a color to apply to the feature if found. Each feature definition in the library is compared against the entire sequence, one by one, and if a match is found, the feature name and formatting defined in the library are applied to that part of the sequence. Thus, raw sequences can be rapidly converted to a table of feature names and base ranges. This modular approach benefits both the data sharing as well as the data preservation roles of ApE. Feature libraries can be exchanged between lab members or between lab groups. For example, collections of PCR primers can be stored as feature libraries and used to annotate any number of sequence files.Because feature visualization is so important, ApE provides three ways to see what features are assigned to a piece of text. First, placing the mouse pointer over any character displays the feature names of all features assigned to that character . Second,Finally, features can be hidden from the current display without deleting the feature from the feature table. This modular approach allows the user to visualize features in many different contexts.ApE is a sequence editor, and contains powerful general and DNA specific text editing tools, including basic text input, sequence search, ORF search, specialized copy and paste functions, and brief instantaneous analysis of selected text.ApE’s main sequence window resembles many classic text editor windows, except that it is limited to representing DNA bases: either ACGT, ACGTN, or IUPAC degenerate base codes. The sequence can be linear or circular, as specified with a button at the top of the window . In circApE has a search function specialized for the needs of molecular biologists. The find window, accessible from the main menu “Edit > Find,” or from the magnifying glass icon on the toolbar, has a basic text input. However, the search can be specified to find DNA sequences using the search input as degenerate bases, single letter amino acid codes, or literal bases. Depending on the setting, the character “N” would match any single DNA base, the asparagine codons AAT or AAC, or just the character N, respectively. Further, the search can be specified to match just the top DNA strand, or can search for the match in both strands, and can match the characters in a case-sensitive or case-insensitive search. Finally, for DNA searches, the user can allow a fixed number of mismatches to occur between the search string and the sequence, or can specify only a fixed number of bases at the 3′ end of the search be required to match.In addition to a text-matching search function, ApE has an open-reading-frame-based search function. This search can find the next or previous open reading frame relative to the current insertion cursor. A user can filter ORFs requiring a minimum length, requiring starting with a methionine or the next codon after the next stop, and requiring the ORF to be on either the top or bottom DNA strand. These settings are quickly accessed in the “ORFs” menu.via the “Edit > Copy Special” menu. First, the function “Copy all as GenBank” will copy the entire sequence together with the associated header and feature records as a plain text version onto the clipboard. This can be used to make a complete record of the sequence in a lab notebook, an email, or a laboratory database, for example. These clipboard files can be re-imported and will open as a new sequence window. Second, the functions “Copy Uppercase” and “Copy Uppercase Rev-Com” allows the user to copy discontinuous regions of interest. Third, the functions “Copy Translated,” “Copy Uppercase Translated,” “Copy Translated Rev-Com,” and “Copy Uppercase Translated Rev-Com” allow the user to translate a continuous or discontinuous region for export into protein analysis software. Fourth, “Copy as FASTA” generates a FASTA version of the selected text, with the file name and selection indices in the FASTA header. Fifth, sequences can be copied as NCBI Bankit tables for submission to the NCBI database.Along with basic copy and paste functions, ApE also has many other functions that operate through the clipboard Finally, ApE displays four important attributes of user-selected sequence: melting temperature (Tm), %GC, a representation of the open reading frame  and a trAt times, the user may need other ways of visualizing aspects of a sequence that go beyond the basic feature highlighting in the sequence window. First, the “Text Map” function, available from the menu item “Enzymes > Text Map” or from a toolbar icon, allows the user to generate a customized text formatted representation of the sequence that includes multiple data tracks. Data tracks include restriction enzyme recognition sequences, position index, translation, bottom strand sequence, and feature regions. The user can then copy or save the window as a plain-text or rich-text (RTF) representation for archiving, sharing, or presenting the sequence. The “Translate” function generates a translation of a selected sequence region or CDS-type feature of a sequence. The translation can be formatted as single or three-letter codes, with optional spacing, line numbering and corresponding DNA sequence. The analysis includes the number of translated amino acids and the predicted molecular weight of the protein.Second, the “ORF Map” function, available from the menu item “ORFs > ORF Map” generates a simple visualization of ATG start codons and amber, ochre and opal stop codons in all six frames of a sequence region. In order to aid in visualizing the most potentially relevant open reading frames, the user is given the option of specifying a minimum cutoff for highlighting regions between stop and start or between adjacent stop codons. The user can then click on any highlighted region to select the corresponding region of the parent sequence.Third, the sequence can be visualized using the “Graphic Map” function, available from “Enzymes > Graphic Map” or from the toolbar. This function converts all of the sequence features and selected enzyme cut sites into a vector map . The mapEach graphic window can be saved into four vector-based file formats: encapsulated postscript (eps), scalable vector graphics (svg), XML-based Power Point (pptx), and portable document format (pdf). By exporting into four different popular vector-graphic formats, ApE visualizations can be imported into many other programs that can represent vector graphics. For example, sequence maps can be read into Inkscape, Adobe Illustrator, OpenOffice Draw, or LibreOffice Draw for writing papers or lab reports, or posted directly to a website as svg for sharing on the web. Finally, the pptx format can be read into Power Point, Google Slides or Apple Keynote for presentations.E. coli Dam and Dcm methylase sites. ApE maintains a database of overlapping configurations that are not cut. Thus, the user can choose to calculate the number of enzyme sites as though the DNA is or is not from a methylated source.ApE has several tools, described in later sections, that use restriction enzyme sites as input. First, we describe how restriction enzyme sites are selected. The central switchboard for restriction site recognition in ApE is the enzyme selector dialog. Enzymes selected in this dialog become the currently “selected set” of enzymes that can be used in subsequent analysis or visualization tools. The selection dialog presents a central window with a list of enzyme names . EnzymesEnzyme sites can be filtered using the “calculator” function below the enzyme selection list. The calculator works by setting a desired number of sites in the current window, as well as membership in an enzyme group. The enzymes that meet both the number and group membership filters are previewed as underlined in the selection list. The user can then choose to apply one of three selection operations: First, “Select” will add all of the underlined enzymes to the current selection. Second, “De-select” will remove all underlined enzymes from the current set. Finally, “AND” will select the intersection of the current set with the filtered set.Because the enzyme selector serves as a central place to select enzymes that are used in other tools, the dialog supplies a shortcut to several of these tools as a convenience. These appear at the bottom of the enzyme selection dialog. For example, the “Highlight” function will highlight the recognition sequences of the selected restriction enzyme set in the selection. In the X-ray window, these highlighted enzyme sequences show not only the recognition site, but also the cut sites, as a small tick mark upwards at the position of the top strand cut, and downwards for the bottom strand cut.The “Digest” function, available from the “Enzymes” menu or the toolbar, generates a simulated agarose gel that would be produced when the selected sequence is digested with the selected enzymes . PlacingWhile simple single-lane or multiple-lane-single-digest gels can be generated from the selection dialog, more complex simulated agarose gels can be generated in a single step using the “Digestion Dialog”, available from the “Enzymes” menu or from the toolbar. In this dialog, each gel lane is represented by a row. Each row is either a DNA or ladder. Each DNA row can then be digested with single or multiple enzymes by activating a checkbox representing the specific enzyme column in the row. Partial digests can be accomplished by activating the "%" button in the enzyme selection region. Each enzyme can then be digested between 0 and 100%.For some applications it can be useful to identify sequences that can be mutated to generate a new restriction enzyme recognition site. ApE has two functions that do this kind of analysis. First, “Silent Sites” examines the currently selected region and identifies potential sites that maintain the reading frame. Second, “Add Diagnostic Site”. identifies new recognition sties independent of reading frames. Instead, it allows the user to specify a maximum number of base changes allowed for the generation of the site. In both analysis results, the sequence is live-linked to the parent sequence, so that clicking on any base representing a base in the sequence selects that base in the parent window.ApE includes simulators for the classic restriction-ligation reaction, Golden Gate assembly, Gibson assembly, Recombinase assembly, and PCR. These are available from the main menu “Tools”.via restriction digestion followed by DNA ligase. The ApE tool “Restriction-Ligation Assembler” is able to simulate this reaction with one to three DNA fragments . The ApE “Recombination Assembler” tool  functionC. elegans RNAi using an LR-recombination reaction. ApE allows users to design new reaction prototypes to accomplish any DNA recombination reaction in silico. The “Recombination Reaction Editor” dialog allows new prototypes to be created If a user wants to design recombination reactions involving a different recombinase or integrase, new recombinase reaction sites can be added using the reaction editor.Although most users only perform the basic Gateway reactions as described in the product literature, the Gateway recombination reaction is very flexible and has been adapted to specific use cases. For example, the pHELLSGATE plasmid is designed to use a PCR fragment containing attB1 and attB2 ends to recombine simultaneously into two attP1-attP2 sites to generate a hairpin silencing construct for use in Arabidopsis. Similarly, pWormgatePro allows users to build a similar hairpin construct for Amplification by PCR is the most critical method in molecular biology laboratories. The ApE tool “PCR Reaction” uses PCR primer databases, searches sequences for primer binding sites, and simulates a PCR reaction with a given set of primers . A majorBinding is determined by a user-defined minimum annealing length, starting from the 3′ base of the primer. The user can allow up to ten mismatches in the primer while still requiring the minimum number of matching bases, and define the maximum number of bases that can be mismatched specifically at the 3′ end of the sequence. By default, the tool then shows a list of primers that match at a single site within the template. The list has data columns: primer name, primer sequence, binding direction, distance of the 3′ end from the current selection, location of the 3′ end in the template sequence, melting temperature of the template-matching region, melting temperature of the full primer sequence, number of mismatches to the template (not including any 5′ extension), and number of matches to the template. Primers in the list can then be selected or deselected by the user. Selected primers are shown in their approximate location and direction on a mini-map diagram in the center of the window. Finally, the user can activate a “Select Primer Pairs” selection mode, which limits the current selection to two facing primer pairs. These primers can be used to generate a new linear sequence window containing the simulated PCR product.In addition to working with existing primer sequences, ApE can assist the user in designing new primers. First, the user can simply select a region of text while noting the melting temperature of the sequence. Alternatively, there is a “Find Primers … ” tool that scans the selection for regions that meet a user-defined minimum and maximum length, melting temperature, percent GC, and 3′ GC clamp. Oligonucleotide primers can be filtered to exclude self-hybridizing sequences, adjacent hybridizing bases and 3′-end hybridization. If the user enters a target primer sequence that the primer should be compatible with, the filter will also apply a cross-hybridization filter on the same parameters as well. While a good first pass filter for primer binding sites, the tool is not as thorough or as flexible as a dedicated primer finding algorithm like Primer3 .In most molecular pipelines DNA must be analyzed to determine if the actual DNA matches the conceptual sequence. ApE can display Sanger sequencing reads and align them to a reference sequence using the “Align Sequences” tool . The aliWhen working with DNA from a genomic source, users will often want to identify single nucleotide polymorphisms (SNPs). A Derived Cleaved Amplified Polymorphic Sequences (dCAPS) PCR assay is a popular way to genotype SNPs. In simple terms, a dCAPS assay uses a PCR primer that has a small number of base differences from the template near the 3′ end adjacent to the polymorphic base. The primer-induced mutations are designed so that products generated from one allele have a restriction site while the other allele is lacking the site. Typically, one allele generates a 250 bp band, while the other allele generates a pair of 200 and 50 bp fragments. ApE has a “dCAPS calculator” tool to quickly design dCAPS primers. The user simply selects the base in the reference sequence that differs in the polymorphism. The tool asks the user to identify the alternate allele base, and then leads the user through the design of dCAPS primers.via the web  is a critical resource to molecular biologists. ApE provides a direct internet link to two NCBI resources: BLAST and Entrez query. The Basic Local Alignment Search Tool (BLAST) , provide the web  and retuMany molecular reactions require DNA in specific molar ratios. For example, bimolecular ligations are often performed at a 3:1 insert to vector ratio. Since the input DNA is usually different lengths and at different concentrations, molecular biologists often need to repeatedly calculate the molarity of different DNA solutions. ApE includes a molecular ratio calculator which can store the molar ratios required in frequently used reactions and can calculate the required volumes of each DNA fragment, given the sequence length and measured concentration.Finally, ApE has several tools that provide functions that may be useful to a more limited user base. The “sgRNA Analysis” tool allows a user to screen sequences against a set of rules  for imprhttps://jorgensen.biology.utah.edu/wayned/ape/, but it is not distributed as true open-source software. It is distributed with most of the rights of open-source software, including rights to inspect and modify the source code. However, no right is granted to redistribute modified source. The Tcl source code is archived at https://github.com/mwdavis2/ApE. In the event that a community member wishes to propose a code addition or bug fix for incorporation into the centrally distributed version, they may initiate a pull request and contact the author to agree to a contributor license agreement. This key distinction from true open-source projects was made consciously to prevent project forking which we feel is inevitable with redistribution of modified versions. For small projects, especially projects lacking a user interface, forking can be a benefit. Each new user can customize the software to meet their needs, and new users not interested in modifying the source can chose from a wide variety of flavors of the code that might meet their needs. However, with a more complex program involving a user interface, forking runs three risks: 1) diluting the user base among multiple incompatible versions; 2) introducing inconsistent and unexpected user interface elements; and 3) having inconsistent quality checks on different versions of the software. With a single distribution source, there are no competing versions. Further, there is a single decision point for any aesthetic choices in the user interface. Finally, all bugs are referenced to a single version and can be corrected without needing to distribute potentially incompatible patch files to fix bugs that may or may not exist in different distributions. However, one of the strong suits of ApE is that many of its features and user interfaces have been inspired by input from users requesting new or modified functionality.ApE is distributed free of charge and can be downloaded at


Gender has been found to influence attitudes towards animals, with women demonstrating more positive attitudes than men in some countries. As attitudes determine consumer behaviour, to a certain extent, and China  has witnessed major social changes in recent decades, we conducted a survey to investigate whether gender and age influenced attitudes towards animals. Respondents self-classified their gender as female, male, other, or they did not disclose it. We found that the attitudes were determined by a combination of gender and age, with more support for animal welfare in women aged 18–24 years than in older men (25–54 years). Those that did not disclose their gender and those declaring it as ‘other’ appeared to have different attitudes to those declaring it as female or male.A person’s gender and age can influence their attitudes towards animal welfare, with more benign attitudes generally ascribed to women. Given that attitudes influence consumer behaviour and the rapid recent social development in China , we surveyed over 1300 individuals across China to elucidate the role of gender and age in determining attitudes towards animals. Respondents self-identified their gender as male, female, other or not revealed. There were interactions between age and gender for many of the survey items, demonstrating that the effects of gender were dependent on the respondents’ age. Women aged 18–24 reported more benign attitudes towards animals than older men  and more empathetic responses were found in young respondents generally, although this did not necessarily translate into a willingness to pay more for higher-welfare animal products. We propose, drawing on Social Identity Theory, that women see animals as part of their social group, whereas men tend not to do this. Those responding as neither male nor female, i.e., as another gender, and those not revealing their gender appeared to have different relationships to animals than those responding as men or women. It is concluded that within Chinese culture, attitudes towards animals and their welfare are complex and influenced by an interaction between gender and age. Large differences in attitudes towards animals have been recorded, arising from characteristics such as gender, age, education level, field of work, religion, culture, and nationality ,2,3. In Gender differences in attitudes towards animal welfare topics have been demonstrated across several different countries and cultures. In the USA, women are more concerned with animal protection than men are . SimilarGlobal differences in the attitudes of women and men towards animals may be explained by a variety of factors, such as political and cultural influences and the level of dependence of women on men. In a cross-country study across Eurasia, women only tended to report greater concerns for animal welfare than men in countries with a high level of gender equality . This haGender differences in attitudes and behaviours toward animals are not immutable. They tend to develop during or even before adolescence ,16. In pAge can also be an important factor that influences attitudes towards animals. In their review, Ormandy and Schuppli  noted thThese differences in attitude resulting from age are evident in diverse cultures. Young people in China have more positive opinions towards animals than middle-aged and older adults , in thatIn a recent study, Carnovale et al., (2020)  identifiIn the current study, data from the same questionnaire as used by Carnovale et al.  were useThe questionnaire and survey method were approved by the Human Research Ethics Committee of the University of Queensland, Australia (#2019001811) and have been previously described in full  Appendi, Box A1.The current paper focuses on the effect of two of the most significant demographic factors—age and gender—which are recognised to be influential on attitudes towards animals. The effects of further demographic variables will be described in subsequent papers. Survey respondents were asked to self-identify their gender by selecting one of four categories: Male, Female, Other, and ‘I prefer not to say’ . We used the term ‘gender’ here, rather than ‘sex’, because the former refers to social and cultural differences in identity, expression, and experience, rather than sex, which is the biological characteristic recorded at birth  . We did Response variables that were structured as ordinal categories were transformed to a numerical scale from 1 to 5 in order, from negative to positive affirmation, for example, 1 (not sure) to 4 (many times); 1  to 5 (to a great extent); 1  to 5 (extremely); 1 (definitely not) to 5 (definitely); 1 (very poor) to 5 (very good); 1 (much worse) to 5 (much better); 1 (strongly disagree) to 5 (strongly agree); and 1  to 5 (very important). A six point scale (1 (5%) to 6 (>100%)) was used to ask how much extra people would be prepared to pay for high welfare. In the case of binomial categories, the data were transformed into “1” for the higher number of responses and “2” for the lower number of responses.p < 0.05), results for a model including the relevant interactions are provided; otherwise, results for the relevant models without interactions are provided. For binary and nominal dependent variables, Binary and Nominal Logistic Regression models, respectively, were used. Data are displayed as the mean response on the relevant ordinal scale and the number of responses for each group , Non-revealers; and Age: 18–24, 25–34, etc.) as relevant. As some gender categories had only a small number of responses, they were not able to be included in the interaction models. These were the non-binary gender category and the two oldest age groups, 55–64 and ≥65 years.All analyses were conducted using the statistical package Minitab 18.0 . Descriptive statistics were generated and have been previously reported . The effIn total, 2170 people were approached to participate and, of these, 1301 completed the questionnaire. Overall responses have been reported previously in Carnovale et al. ; howeverRespondents almost equally self-reported as male or female: 47% and 48% . Of the remaining participants, 3% (n = 39) identified as Other (non-binary) and 0.5% (n = 7) said that they preferred not to reveal their gender . For most gender categories, participants were most commonly between 18 and 24 years of age  . For theWomen that were younger did advance more pro-animal welfare opinions than the other groups. In particular, women aged 18–24 (n = 189) were more likely to say that they lived in harmony with animals compared to men aged 35–44 years (n = 114) or non-revealers aged 25–34 years (n = 10) . They weCompared to young women (18–24 years), non-revealers  were more likely to say that animal welfare was worse in China compared with other countries, although the sample size was small. Both groups were generally in agreement about the importance of caring for a range of different animals. Importance scores were significantly higher for the group of young women than the non-revealers, for the animal categories of mammals, reptiles, birds, and stray animals ; howeverIn comparison to either men or women, non-revealers indicated they had learned about animal care mainly from family and friends, more commonly than either men or women . Of non-Participants who were non-revealers reported the standard of animal care in China as poorer than the other gender categories did, and they were more likely to report it as being worse than other countries . In termIt makes me feel good”, “To improve profit from animals”) with a higher score indicating stronger agreement. There were few differences between gender categories for these statements, with the exception of “For food safety” where non-binary participants reported a lower agreement with this statement than other groups, and “For the sake of the animals” where the highest importance score was given by men. Gender differences were also apparent in attributing importance to specific conditions provided for animals, namely “A comfortable environment”, “Opportunity to perform natural behaviours”, “Absence of fear or distress”, and “Absence of pain”. Participants from all gender categories gave positive scores for these on average, indicating a belief that these are important; however, scores from non-binary participants were typically lower than for the other groups . People from all gender categories were largely supportive of these measures with mean scores of 3.29 or higher, on a 5-point scale from 1 (Strongly disagree) to 5 (Strongly agree). However, these were supported the least by women . ParticiMost of the respondents of all the gender groups declared that they had never heard of animal welfare, but they thought it was extremely essential to care for animals, and perhaps having the possibility to learn about this topic at school would be beneficial. All gender categories felt society was responsible for animal welfare. Pets, mammals, and birds were the animal taxa on which all genders agreed on their importance. For the sake of the environment and human health, the majority of all gender categories considered it important to care for animals. All respondents indicated they care about animals for religious reasons and to make themselves feel better. Increasing profits and increasing the quality of animal products were similarly important to the respondents, regardless of their gender. Irrespective of the gender of the respondents, it was thought to be important that animals be able to exercise for physical fitness and have control over the environment in which they live. It was also thought to be crucial to have enough area and access to drinkable water and prevent contracting diseases or injury. The management of animals was declared important by the majority of all gender categories, especially concerning transportation time and slaughter management, and they all thought it extremely important to have an animal protection organisation.Many age-related differences were found in participant attitudes towards animals. The youngest respondents (aged 18–24 years) were more likely to have heard of the phrase “animal welfare” than those aged 65 years or older. They were also more likely to report that they lived in harmony with animals, and that caring for animals was important to them as a person, in comparison to several of the older age groups   but not The youngest respondents were more likely to believe that all of society is responsible for animal care than those aged 45–54 years . They weAge effects were evident for participants’ views on why animals should be cared for, and while these effects were mixed, in general, younger respondents were slightly less likely to agree that animals should be cared for because it made them feel good, for the sake of the animals, for religious reasons, or to improve profit, than those in older age brackets . The youSome factors were consistent across age groups. The age of respondents had no effect on their judgment of animal welfare in China as “poor” and, compared to other countries, they agreed that it was “somewhat worse”. In all taxa animals listed in this survey, mammals, birds, and stray animals were identified by all respondents of any age as important, while wild animals, experimental and agricultural animals were identified as extremely important. Regardless of age, all respondents agreed that the most important reasons for caring for animals were food safety, human health, improving the quality of animal products, and the need for animals to be free of fear and allowed to exhibit normal behaviour. The reasons given for the care for animals are that the animals need to have the possibility for physical fitness and control over their environment. Animal care and management was found to be very important regardless of respondent ages, particularly in terms of transit time and slaughter management, but it was also thought to be highly necessary to have an animal protection organisation and law that protects animals.Previous surveys, mostly based in Western countries, have identified differences between men and women in their opinions concerning animal welfare ,14,15,22In general, respondents who were non-revealers n = 39) expressed greater sensitivity to animal welfare on most questions, and their views differed from both men and women. Attitudinal differences between men and women have been reported across many contexts in relation to animal welfare 9 express,6,7,8,9,In the current study, gender was not limited to male and female, but also included options for participants to self-identify as a different gender  and to refrain from disclosing their gender . In the past, surveys have often used exclusively binary gender categories, which do not allow adequate representation across this influential demographic variable. Methodologically, and ethically, surveys should include multiple response options for gender  as gendeParticipants in the current study of different ages and genders were asked to what extent they live in harmony with animals. To live in harmony with animals and nature is an important traditional principle in China that predates even Confucianism . It is eYounger respondents (18–24 and 25–34 years) and those in the oldest group showed the most reluctance to pay more for products from animals that were better cared for. This may be reflective of their relationships with animals but is also likely to be influenced by their financial resources or purchasing patterns. Household income is known to affect purchasing decisions in China for other ‘premium’ products such as organic food, although these issues interact as animal welfare is also a motivator for the purchase of organic products in China . GeneratIn the current study, middle-aged male participants typically had less positive attitudes towards animals than younger female participants. This gender difference aligns with previous research conducted internationally that also found more benevolent attitudes in women towards animals, particularly in regions with a high level of gender empowerment . HoweverIn our study, only a small number of participants chose not to reveal their gender; however, those in this category expressed high levels of sensitivity to animal welfare for many survey items, and on several aspects their views differed from the other gender groups. Non-revealers were more likely to say that they learned about animal care from family and friends, but no difference was found for other sources of information, such as formal study. They were more likely than the other gender groups to say they lived in harmony with animals. In contrast, they were less supportive of looking after animals for the animal’s own sake than other groups, and less supportive of doing so for food safety reasons than men or women.Failure to reveal gender may occur for several reasons. In our case, non-revealers may perceive themselves to fall outside of the conventional binary assignation of gender roles. Previous research suggests that non-revealers to gender identity questions can occur for several reasons, including participants being unsure of their own gender at the time, undergoing gender transitioning, or discomfort with the other options given . HoweverWe anticipated that non-binary participants in this study would demonstrate more benevolent attitudes towards animals, as previous research suggests marginalised members of society often develop strong attachments to companion animals, e.g., ,59,60, aThe number of non-binary respondents in the current survey was very low in comparison to those who identified as male or female; therefore, these results should be considered preliminary until they are more systematically explored in future research. Imbalance in participant numbers for gender and age is a limitation of this research; however, this study provides important insight into the influence of these demographic variables on attitudes towards animals in China.Another potential limitation of this work is that in mainland China, the English words “gender” and “sex” are translated to the same word (性别) and official documents generally only allow for two categories , which may have influenced how participants responded to this question in the survey. Some misunderstanding of the translated version of the questionnaire may have occurred, which could be a limitation, although this was not reported by the questioners.This study provides evidence that the attitudes of the Chinese population, and relationships between these attitudes and age and gender, are not greatly different from those recorded elsewhere; the sensitivity of the Chinese population to animal welfare does not differ much from the rest of the world. The identification of gender and age groups that are more concerned with the welfare of animals confirms the value of, and interest in, educating the young in the formal school setting on issues of animal welfare. Whether this interest is related to the age of the respondents, where we might expect that their attitudes would harden as they get older, or if it is a generational factor, with continued interest into later life, could be confirmed by a longitudinal study, ideally asking the same respondents in later life, but this is beyond the bounds of this study.This study investigated the effect of gender, age, and their interactions on Chinese respondents’ attitudes to animals and their welfare. While age was influential on survey responses, the effect was not linear. Young women tended to have more empathetic attitudes towards animals than men aged between 25 and 54 years. Similarly, young participants had more benevolent attitudes towards animals than middle-aged participants; however, they aligned in many aspects with the oldest age groups. Gender also influenced survey responses, with non-binary participants generally indicating less positive attitudes towards animals and their management, and those who chose not to reveal their gender indicating more positive attitudes. However, these results should be considered preliminary as only a small proportion of respondents chose gender categories other than male or female.


A patient is vulnerable and exposed due to illness, relies on and surrender to other people. In caring this means a special dependency. The aim of this study was to describe the origin of the concept ‘exposed’, to elucidate how the definition of this term has changed over time, and to outline its relevance in caring science.A semantic concept analysis in two phases was conducted.The findings show that over time, the degree of binding of the following synonyms has become stronger; unprotected/defenseless, naked/bare, pressed/jostled and unsafe/unprotected and can easily be transferred to a clinical nursing context.This study provides a deeper understanding of the concept 'exposed' semantically. Being exposed is a profound experience for patients who need to be seen as the person they really are. A life-world led care has an existential power that can support professionals, strengthen patients’ health processes, and alleviate the patient’s suffering. The term exposed (utsatt) is used in the Swedish language in various ways, e.g., for people in vulnerable positions.Being human includes existential vulnerability. Temporality and instability in our lives, along with the inability to control our closest living conditions, are a basis for experiencing both fragility and vulnerability in existence Arman, . To be hCaring ethics is the basic approach towards the patient and the care, regardless of the ethical guidelines that exist for each health care profession. Human dignity, meaning the right to be confirmed as a unique person, precedes everything else. Suffering raises the ethical requirement for care  x 100/possible binding + possible bindingFor example, exposed (utsatt) is bound to the synonym perilous (riskabel) in four of the eight dictionaries; perilous (riskabel) is bound to the concept exposed (utsatt) in two of these dictionaries. The concept exposed (utsatt) is an entry in all eight dictionaries; the synonym perilous (riskabel) is an entry in four of these dictionaries. The degree of synonymy is calculated according to the formula: (4 + 2)×100/8 + 4 = 50%.All research, empirical or theoretical, raises ethical questions. By striving for as open and well-founded argumentation as possible, we have strived to follow the directions for good scientific practice, claiming both transparency and stringency , 2013. SThe results start with an etymological and historical description of the concept followed by a matrix phase where dimensions of meaning and synonyms are presented. The paradigm phase shows how the different synonyms are related to the concept, and the concept’s change over time is described.exposed separately, but refers to the hypernym “unfree/not free” where, for example, exposed to and vulnerable are listed as synonyms , in its adjective form, existed in most of the selected dictionaries in three different meaning dimensions .Table I“Which can easily be attacked” and “which is a target of attack” are two expressions that the dictionaries specify as synonymous for exposed (utsatt). However, the word attack does not appear as a synonym. This meaning shows that the person who is exposed is passive and exposed by someone else. It is interesting to note the expression “put yourself at risk” . Thus, iprekär), perilous (riskabel), threatened (hotad), dangerous (farlig), vulnerable (blottställd), unprotected/defenceless (oskyddad), naked/bare (blottad), pressed/jostled (trängd) and unsafe/unprotected/exposed (exponerad).In the concept analysis, synonyms from the dimension “in an exposed situation”, with at least two bindings to the concept were included in the first calculation of degree of synonymity, and synonyms with at least a 27% degree of synonymy were included in the subsequent analysis. The synonym “threatened” had only a 22% binding. However, this synonym was chosen in the 2004 analysis, with a 30% binding, and therefore it was also included in 2019. The synonyms are: precarious (In the discrimination paradigm  the synoPrecarious does not appear as a synonym in dictionaries published after 2006. The one synonym that is stable regarding its degree of binding is Dangerous. Precarious, Perilous and Threatened have weakened. Synonyms that have become stronger bindings are unprotected/defenceless, naked/bare, pressed/jostled and unsafe/unprotected/exposed (/exposed .Table Vin an exposed situation, since this meaning is most related to clinical caring practice. This may be seen as a limitation. Perhaps a broader view of the concept might have given a different understanding but our results show a robust description of the concept related to caring. To ensure a robust analysis of the concept exposed and determine its relevance for a caring science, a semantic analysis was conducted to get a sense of the meaning of the concept. To elucidate how the term might have changed over time, the analysis was made twice, with a 15 year gap in time. In the analysis from 2004, only printed books were used. Since then, digital sources have become available. Thus, in the second analysis, digital as well as printed sources were used.A concept analysis gives definitions to the use of a concept in both theory and research and can also help clarify terms in nursing that have become catch phrases and hence either have lost or changed their meaning (Eriksson, Koort Koort,  states tThe choice of language may be considered as a limitation. However, Sivonen et al (Sivonen et al., exposed had become stronger and more stringent in the meaning dimension in an exposed situation.Over the last fifteen years, the concept’s relevance for clinical caring practice has strengthened. The degree of binding of the synonyms has become stronger for those synonyms that easily can be transferred to a clinical nursing context i.e., unprotected/defenceless, naked/bare, pressed/jostled and unsafe/unprotected c.f. . In the A semantic analysis can be further developed by connecting theoretical and clinical caring science from specific or more general caring contexts. The results can thereby create a deeper understanding of the data, support the analysis and further develop the understanding of the concept and the experiences in a caring context (Eriksson, vulnerable and exposed due to illness. This vulnerability also affects the next-of-kin (Dahlberg et al., Overall, the findings  can be rnaked and exposed. Being exposed means being unprotected as a human being in a vulnerable situation (Todres et al., From a caring science perspective, our findings showed that the concept exposed needs to be highlighted, as it entails patients suffering both from poor health and from the care they receive or do not receive, which in turn can be related to loss of dignity. Objectification of human beings in health care settings is an aspect of being vulnerability raises the human ethical obligation to care for a unique person. The power lies in the hands of carers or of the health care system that risks the patient’s dignity. Patients can be denied alleviation of their suffering as persons and human beings, and feelings of worthlessness can arise. The power is related to lack of knowledge, the patients’ insecurity and not being understood. This exposedness means a vulnerability that can be experienced as overpowering and annihilating (Dahlberg, Maintaining human dignity means maintaining the right to be confirmed as a unique person. The presence of suffering and vulnerability, exposedness, finitude, frailty, unprotectedness, insecurity etc. According to Zaner (Zaner, vulnerability, being pressed/jostled, violence, and power, leading to precarious situations for the person’s own being that is being threatened.There is a strong relationship between existence and caring. Carers ought to become much more attuned to the importance and value of the experience of the person who is suffering (Galvin & Todres, r Zaner, , clinicadanger and in need of restoration. Alas, some degrees of rupture are very difficult, perhaps impossible, to restore (Galvin & Todres, Suffering caused by care is the suffering that the health care providers cause to the patient by violating the patient’s dignity, for example exercising power or failing in their care (Eriksson, Cubellis (Cubellis, Exposedness is a profound experience for patients. This concept analysis can help in deeper understanding a person’s unique exposed situation and adds to the knowledge base of exposedness within nursing care. Life-world led care has existential power that can support professionals in strengthening patients’ health processes. By this is meant finding balance as opposed to being exposed and experiencing human vulnerability. This balance alleviates the patient’s suffering and reduces the risk of suffering by care. Since caring science as a science is not connected to one specific profession, everyone who meets exposed human beings in their daily work, e.g., all health care professionals, teachers, policemen, social workers, can use this knowledge to further understand exposedness from a human perspective.


Ion transport under nanoconfined spaces is a ubiquitous phenomenon in nature and plays an important role in the energy conversion and signal transduction processes of both biological and artificial systems. Unlike the free diffusion in continuum media, anomalous behaviors of ions are often observed in nanostructured systems, which is governed by the complex interplay between various interfacial interactions. Conventionally, nanoionics mainly refers to the study of ion transport in solid‐state nanosystems. In this review, to extent this concept is proposed and a new framework to understand the phenomena, mechanism, methodology, and application associated with ion transport at the nanoscale is put forward. Specifically, here nanoionics is summarized into three categories, i.e., biological, artificial, and hybrid, and discussed the characteristics of each system. Compared with nanoelectronics, nanoionics is an emerging research field with many theoretical and practical challenges. With this forward‐looking perspective, it is hoped that nanoionics can attract increasing attention and find wide range of applications as nanoelectronics. Nanoionics, as an analogy of nanoelectronics, refers to the transport phenomena, functions, and applications of ions that are confined in the nanospace. This review introduces the concept, recent remarkable advances, and insight on the current challenges of nanoionics, which can be categorized into biological nanoionics, artificial nanoionics, and hybrid nanoionics. Based on semiconductor physics, nanoelectronics has become a crucial part of electronics and is of predominant significance in facilitating human technological progress, particularly in the information technology and energy fields. However, as the dimension decreased to sub‐5 nm scale, higher‐performance solid‐state nanoelectronic devices face physical limitations and performance challenges. For example, the speed and energy efficiency of silicon complementary metal–oxide–semiconductor (CMOS)‐based computing hardware are approaching to its theoretical limit. Meanwhile, it is widely believed that nanoelectronics based devices could hardly be comparable to the human brain in terms of high‐level information processing, such as pattern recognition and intelligent reasoning, especially considering the energy efficiency. One of the major reasons lies in the distinct working principle of the communication media between solid‐state nanoelectronics and soft biological nanoionics systems. Cell‐to‐cell communications in an intelligent organism is expressed in the language of various ions and small molecules, in contrast to the mere flow of electrons. Motivated by this observation, the utilization of ions as the charge carriers has emerged as a new research frontier, and received increasing attention.The advancement of nanotechnology over the past several decades has allowed the engineering of material structures across multiple length scales.1, 2 An organism is a typical and ideal model of ionic circuits whose function is based on the interplay between pre‐designed structures of biomacromolecules and various ions of different charge, size, and mobility. Taking the inspiration from organisms, the new generation of artificial intelligence systems, including sensory devices, energy devices, and human–machine interfaces that work in a nanoionic way, would possess several key advantages. For example, they could have much lower power consumption than conventional processors, and exploit integrated nonvolatile memory using biocompatible or biodegradable logic circuits for sensing. Additionally, these devices could be explicitly designed to support dynamic learning of complex and unstructured data. Moreover, as exemplified inbatteries and fuel cells, ions are also important for the transformation of chemical energy to electrical energy. Therefore, the significance of ionic charge carriers is by no means less than that of electronic charge carriers.In Nature, the transmission and processing of information, as well as the energy conversation and storage are often mediated and regulated using ions and fluids transportation at small scales.8 An o Beyond that, it is believed that ionic charge carriers have broad applications, especially in designing information processors, thanks to the specific characteristics of ions, such as valences, sizes, and polarizabilities. For example, various ionic flows including Na+, K+, and Ca2+, determine electrophysiology in biology. As a result, like biological intelligence organisms, artificial systems that run on ions should have the potential to be more efficient than today's electronic devices. Nanoionics based on multi‐ionic carriers, analogous to nanoelectronics, can emulate biological functionality more directly and can also be used as an emerging technology that bridges artificial intelligence systems and biological systems seamlessly.In recent years, research using ions as charge carriers has received increasing attention in several specific research fields, e.g., electrochemical energy conversion and information storage.16 BeyFigure As outlined in 2 Later, Maier and several other scientists invested enormous efforts to shape this research topic and made a significant contribution to it. Originally, the concept of nanoionics referred to the study of ion transport in all‐solid‐state nanoscale systems. However, nanoionics is also related to ion‐transport and storage phenomena in liquids; therefore, the concepts of nanoionics should be much broader and more general. For instance, the materials of choice should not be restricted to nanostructured solid‐state materials, rather they should be generalized to systems, such as liquid and biological materials, as well as the hybrid of the two. Similarly, the application of nanoionics may not be restricted to typical electrochemistry‐related fields, but also include processes such as energy and information transfer  and electrostatic forces increase with decreasing distance between the surface and the particle . They can generate high voltages of up to 600 V and currents of 1 A when they are attacked by their natural predators. Na+ and K+ channels in the cell membrane that open and close periodically are the basis of their specialized electric organs. Benefiting from its relative permeability to Na+ and K+ ions, a single unit of charge‐generating organ can generate a small action potential of ≈+65 mV, and 1000 units connected in series and in parallel resulting in the simultaneous high voltage and high current. Some bacteriorhodopsin transforms solar energy into adenosine triphosphate (ATP) by light‐driven ion pumps, which can pump ions (protons) from low to high concentration to build an electrochemical potential gradient which powers the ATPase for ATP synthesis. In the mammalian brain, the distribution of calcium ions in dendrites may represent another essential variable for the processing and storing of information. Calcium ions enter the dendrites through voltage‐gated ion channels in the cell membrane, which results in rapid local modulations of calcium concentration within the dendritic tree.Precisely controlled ion transport across cells and organelles are crucial for life. In a biological neural system, ion channels are the molecular basis for all the electrical activities in initiation, processing, and transmission of information. In single cells, ion flows are involved in the regulation of a wide range of processes from signaling, action potential, pH balance, volume regulation, to and the cell cycle. In higher forms of living organisms, they also determine immune responses, fertilization, muscle contraction, and secretion Figure 3. Meanwhile, controlled ionic processes can also be used to directly modify the composition and/or structure of the material itself at the atomic scale, allowing new nanostructures to be built on the fly, in a reconfigurable fashion.Additionally, recent studies showed that more complicated biological effects that are critical for learning and memory can also be emulated by the internal ionic processes during resistive switching, e.g., nonvolatile memories, including resistive random‐access memory, conducting bridge random‐access memory, and phase‐change memory. This enables efficient neuromorphic systems to be implemented using solid‐state devices and networks.56 Mea4.2 In a liquid medium, ions exist as solvent‐free ionic liquids or solvated ions, i.e., ions surrounded by solvent molecules. Different from ion diffusion in bulk systems, ions in nanospace demonstrate very different behaviors. Spatial confinement could constrain the motion of ions and/or modify their interactions with each other and with the nanostructure surface, resulting in some anomalous ion‐transport properties.Liquid nanoionics refers to ion transport in liquid phase that is confined in nanoscale systems, e.g., in nanochannels, nanopores, nanotubes, or porous materials.17 In  Among them, pore size is the most important factor. In this section, ion transport in nanoscale (2–100 nm) and microporous  will be introduced separately due to their different interaction forces. Meanwhile, the effects of other factors including pore conductivity, charging dynamics and pore connectivity will also be introduced briefly.Several factors including pore size distribution, pore conductivity, pore connectivity, charging dynamics, and ion concentration will affect ionic diffusion or the flow of ions in and out of the pores.62 Amo4.2.1Figure  The Debye screening length is inversely proportional to the square root of the ionic strength, and it is generally in the range of 100 nm. However, for solvent‐free room‐temperature ionic liquids, the Debye screening length is very small and the Debye theory does not apply. The phenomena could not simply be explained in terms of Debye screening, and a more elaborate theory must be used, which will be introduced later. interactions between solvent molecules and ions could be affected by the neighboring ions or the nanospace surface, resulting in the restructuring of solvated ions. In general, the restructured solvation shells around ions in nanospace would be thinner than those in bulk. In this case, the average inter‐ion distance could then be reduced. If the interion distance is comparable to or smaller than dimensions of the Bjerrum length , the weak interion interactions, including hydrogen‐bonding and vdW interactions, become dominant. In this case, ion‐transport behavior becomes more complex and includes a range of unexpected solvation‐involved nanoionic phenomena, e.g., ultrafast ion transport.When the size of the nanospace is comparable to the Debye length, solvated ions in nanoconfinement will “feel” the electrostatic forces from the adjacent surface, resulting in regulatable ion‐transport properties. When the dimension of nanopores is further decreased to a comparable size of solvated ions ,66 int This will result in several interesting nanoionic phenomena, e.g., superionic states. More specifically, ion diffusion will be highly modified in subnanometer pores as the pore becomes charged. The self‐diffusion coefficient of ions confined in subnanometer pores will exhibit a bell‐shaped curve as charging proceeds: At the potential of zero charge, ions inside neutral subnanometer pores diffuse very slowly; as charging proceeds, the self‐diffusion of ions greatly accelerates, even becoming a few times faster than that in bulk room temperature ionic liquids; their diffusion finally slows down when the pore becomes strongly charged. These results also highlight the importance of precisely controlling pore size to realize fast ion diffusion in charged pores. Beyond that, it is suggested that pore connectivity, the distribution of pore throats and the charging dynamics are also important characteristics to consider when improving the ion diffusion and optimizing the existing and developing new porous electrode materials for supercapacitor.However, ion transport properties will be different in the case of conducting confinements. It is clear that the ion‐ion interaction follows the Coulomb law in the case of nonconducting confinements. While in a conducting confinement, the interaction energy between two ions decays exponentially because of the screening of electrostatic interactions induced by the electrons of the conducting walls.72 Thi4.2.2 ionic rectification, and ion pump, which are also found in biological nanopores , several factors, including external force, electrostatic force between ions and interface, wettability, and steric effect, will affect the ion‐transport properties. Ion transport exhibits well‐organized and controllable properties, e.g., ionic gating,77 ion such as voltage‐gated biological nanopores. For example, Qian et al. reported a responsive ionic gate with a high on/off ratio up to 17.8 based on azobenzene (Azo)‐molecule‐modified metal–organic framework nanochannels. Azo is one of the most popular photoresponsive organic groups, as an Azo group can undergo reversible isomerization between trans and cis when triggered by ultraviolet irradiation or heat, leading to an unusual open‐to‐closed switching of the channel. Beyond that, many external stimuli, including electric, pH, thermal, magnetism, specific molecules and ions, also can be used as a trigger to activate the ionic gate.Ionic gating means that the nanofluidic system can work as a gate for ion flow, and it can be opened by stimuli,82 suc This is one of the most common ion‐transport properties of biological nanopores, and it has been replicated and studied sufficiently by artificial systems. Xiao et al. reported an ionic rectification system with a rectification ratio larger than 500 by constructing an asymmetric funnel‐shaped system. They modified the cylindrical and conical segments of a funnel‐shaped nanochannel with different molecules, leading to synergic asymmetric structure and asymmetric surface charge distribution. To realize ionic rectification in nanofluidic systems, an asymmetric configuration is crucial, whether asymmetric structure or asymmetric surface properties (wettability or surface charge density) exist. Based on this principle, Hou et al. reported a dynamic curvature nanochannel system with a reversible rectification switch that endows the artificial system with flexible ion‐transport properties. The nanofluidic system with an ionic rectification property can also be used for the unidirectional transport of several charged drugs or cell messengers, which can then be used for smart drug delivery or to study cell commination by avoiding diffusive leakages.Ionic rectification is the phenomenon where the ion flow in one direction is suppressed while it is unimpeded in another direction.92 Thi To realize the ion‐pumping function in artificial systems, some photo‐responsive molecules are introduced into a lipid membrane or porous membrane. Furthermore, nanofluidic‐based, light‐driven ion‐transport systems can work based on three different underlying working principles, i.e., photochemical, photoelectric, and photothermal effects, by creating an asymmetric surface charge distribution. For example, some semiconductor nanostructures can realize the ion‐pumping function directly via ion transport coupled by photoinduced carriers. Xiao et al. reported a carbon nitride nanotube membrane that can achieve pumped ion transport against a 5000 concentration gradient by creating a gradient surface charge density with light illumination.Ion pumping means that ions can transport against a concentration gradient by consuming external energy. In biological systems, ion pumping is used to pump protons from low to high concentration to create an electrochemical potential difference, which can be then used to drive an ATPase to produce ATP.30 To  water desalination, and various ionic sensors, and they have been summarized well by many reviewers.These controllable ion‐transport properties in an artificial nanochannel of size between 2 and 100 nm are associated with many ion‐related applications, such as osmosis energy harvesting,102 wa4.2.3 This phenomenon is especially obvious in the study of electrical double‐layer capacitors (EDLCs). It has been shown that a decrease in the average pore size of carbonaceous materials used in EDLCs can generate an increase in the capacitance of the cell, especially when the size of the pores is commensurate with that of the bare ions of the electrolyte.With decreasing pore diameter, weak interaction forces, including vdW and hydrogen‐bonding interactions, play an increasingly important role in ion transport, resulting in anomalous ion‐transport properties and applications Figure .[107, For example, Gogotsi and Simon found that porous carbon with pores smaller than the size of ions will lead to anomalous capacitance, which then be explained as the superionic state by Kornyshev and co‐workers. Later, Kaneko and co‐workers  showed that Coulombic ordering is reduced when the pores can accommodate only a single layer of ions, which can confirm that ions in such tiny pores are in a superionic state. Some other work tried to explain the anomalous capacitance from the view of energy barriers. For example, Bruin and co‐workers  explained that adsorption energies gradually increase with decreasing pore size and show a maximum when the pore size is slightly greater than the dimensions of the adsorbed ion and the attractive vdW forces dominate the interaction. At smaller pore diameters, the adsorption energy sharply declines and becomes repulsive as a result of geometry deformations of the ion. This phenomenon is particularly noteworthy in ionophobic porous materials. By overcoming the energy barrier of ionophobic porous materials, it is possible to enhance the charging dynamics and improve energy and power densities of supercapacitors.A different explanation has been given in cases of solvent‐free ionic liquids and solvated ions in aqueous solution. For confined ionic liquids, the non‐Coulombic structure formation is made possible by the repulsive electrostatic interactions between co‐ions caused by image charges induced in the carbon walls, resulting in the superionic state.112 Fo This suggests the establishment of an intrinsic energy barrier that forbids ion entry into the channel. However, the energy barrier can be overcome by different external forces, e.g., an external electric field or increased surface charge density induced by illumination.In the case of ions with a solvation shell, the mechanism is slightly different. The hydrated ions must undergo the dehydration process or deformation to be squashed into the channel.120 Th reported an ionic Coulomb blockade phenomenon observed in a single‐layer MoS2 membrane. In their work, the nanopore used was ≈0.6 nm in diameter, slightly smaller than the hydrated alkali‐metal ions. Therefore, the ions must be dehydrated to pass through the pore, resulting in an additional energy barrier that can be overcome by a larger external electrical field. In this case, the current is suppressed at small voltages and starts to increase sharply when the bias voltage exceeds the threshold value. The observation of this ionic Coulomb blockade phenomenon displays quantum‐mechanical‐like features dominating ionic transport, and it will also offer a new platform to explore novel physics in the research areas of both artificial and biological nanoionics.Feng et al.,121 re reported a hierarchical pore architecture of the carbon membrane with a pore size gradient changing from 60 to 1.4 nm, which enables high ionic rectification ratios up to 104 in different environments, including high‐concentration neutral (3 m KCl), acidic (1 m HCl), and alkaline (1 m NaOH) electrolytes. The ultrahigh and stable ionic rectification ratio resulting from the different energy barriers when ion transport in different directions benefits from an asymmetric structure. Similar anomalous unidirectional ion transport was also observed in a MOF membrane.The sub‐nanoscale channels also exhibit anomalous unidirectional ion transport (ionic rectification) if their structure is asymmetric. We introduced ionic rectification in the preceding section, stressing that it is an important ion‐transport property of protein channels and can be realized by creating asymmetric elements. If the pore diameter is smaller than 2 nm, the energy barrier will play an important role in determining the rectification ratio. Xiao et al.,124 re fabricated an atomic‐scale ion transistor using a single flake of reduced graphene oxide with channel sizes being ≈3 Å. Because the channel size is smaller than the hydration diameters of alkali‐metal ions, an intrinsic energy barrier is created that forbids ion entry into the channel. Subsequently, a gate voltage was applied to increase the electric charge density of the walls to change the energy barrier, and ultrafast ion transport, namely, one that is 2 orders of magnitude faster than ion diffusion in bulk water, was observed beyond a threshold voltage. The authors also studied permeation rates of different alkali‐metal ions, in which K+ > Cs+ > Li+, which resembles the selection sequence of biological potassium channels. These phenomena can be explained by different electrophoretic mobilities, meaning that ionic species with different charge‐to‐size ratios spend different times in the channel because the enormous electric fields inside EDLs cause transverse ion distributions that yield charge‐dependent mean ion speeds in the flow.Xue et al.,69 fab In terms of this energy barrier's origin, it can be confirmed that ultimate confinement largely reduces the electrostatic iterations because of the existence of the superionic state. Several other simulations also suggested that the dielectric constant of aqueous solutions is reduced for very narrow confinements, resulting in strong Coulombic interactions among adjacent ions and promoting their concerted movement with decreasing energy barrier. Of course, the detailed mechanism of ions diffusion in such tiny pore is still not clear and remains open to further simulations and experimental investigation. However, it is for sure that ions in such a tiny space show different packing states within larger pores or bulk systems, and exhibit anomalous ion‐transport properties, including superfast transport speed and superhigh selectivity , e.g., the generation and transmission of action potential. Therefore, it is necessary to develop biological–artificial hybrid nanoionics that work on the principle of ion transport. In this case, it is possible to realize direct neuromodulation without electrical to ionic transduction.As introduced above in the discussion of biological and artificial nanoionics, ions as transport carriers can be used to design various energy and information processes both in vivo or in vitro. The question now becomes “Is it possible to fabricate biological–artificial hybrid nanoionics‐based energy and information systems?” Human–machine interaction (HMI) is a typical biological–artificial information‐exchange system considering that most HMIs are based on electron transport.133 Ho However, establishing an active interaction with biological tissues allows a transition from mere recording of biological functions to dynamic interactive control. What is required at this stage is the capability not only to receive biologically encoded inputs to be processed, but also to generate outputs, hence closing the artificial neural circuit.Most of the existing electronic platforms act like passive devices; that is, they are able to monitor cellular activity without providing significant feedback.136 HoFigure + or K+ ions will flow through the transmembrane to generate an action potential post‐synapse. In this way, a biological–artificial hybrid nanoionics system that works in a way similar to that of its biological counterpart is fabricated. Analogously, biological signals can also be transduced to artificial signals by biological–artificial hybrid nanoionics. These two cases correspond to the “read”  and “write”  of biological signals. We can assume that an ionic device integrating these two functions can realize actual human–machine “two‐way” communication.In this case, two different biological–artificial hybrid nanoionics systems are proposed Figure 7: one fFigure  Organic electronic ion pumps, another artificial nanoionics device made using the ionic conductor polymer PEDOT:PSS, were widely used to regulate ion or molecule flow for stimulation of individual cells or plants. These hybrid nanoionics move a significant step forward in biology‐technology interfacing by translating electronic addressing signals, through ionic or molecular signaling, into brain‐stem responses. However, the translation should not be limited to electric signals since other external stimuli, such as light, pressure, and heat, also have the possibility to be translated into ionic signals by integrating artificial nanoionics devices into biological systems. In addition, biological signals can also be understood by hybrid nanoionics. Keene et al. reported a hybrid synapse by coupling an organic neuromorphic device  to dopaminergic cells , which can then realize neurotransmitter‐mediated synaptic plasticity  by ion‐selective membranes to activate and inhibit a nerve Figure 8a.[137y Figure . In thatWe envision that by combining “input” and “output” biological–artificial hybrid nanoionics systems that can not only signal “read” and “write,” but a chemical communication between neurons can also be introduced or repaired.Figure  Meanwhile, as stated above, the traditional idea that ions move slowly has been challenged in several recently published works, especially in sub‐nanoscale space. In such a small space, superfast ion transport is possible, e.g., by Newton's ionic cradle model. Therefore, the energy‐conversion efficiency and energy density of ionic energy systems could be improved. However, these concepts must be verified before they can be used to solve materials and nanostructure problems.Unlike the attractive biological–artificial hybrid nanoionics information processes, hybrid energy processes are largely ignored because all energy‐generation processes emphasize conversion efficiency and energy density, both of which are drawbacks of ion‐based systems because of the speed of slow‐moving ions. We must hypothesize, however, that electron‐based energy systems cannot be used everywhere, e.g., the sustaining and biocompatible energy source for nanorobots in vivo Figure 9. In th6 moletronics, and nanoionics  must always complement it, even though their value to the process is more or less eclipsed by nanoelectronics. As semiconductor technology moves ever closer to the ultimate physical limits for scaling of devices that utilize electrons, several new options based on working principles other than electron transport should be developed. Therefore, nanoionics has the potential to provide the solution for this challenge.In view of the “omnipotence” of nanoelectronics, a question arises, namely, “Why do we need nanoionics?” Before answering, we must clarify that nanoelectronics is not an isolated scientific development process. Nanophotonics,143 moFigure  Another remarkable advantage for heavy carriers is that lighter mass in smaller devices has a limited information‐storage time because of quantum mechanical tunneling, while the use of larger mass will decrease tunneling probability. Meanwhile, ions have a smell and color  and one could harness these supplementary signatures to design more efficient information processors. For example, Robin et al. used theory and simulations to predict that 2D nanofluidic channels can show nonlinear conduction and function as memory‐effect transistors. By incorporating two nanofluidic memristors in an elementary circuit that refers to Hodgkin's and Huxley's model, the neuromorphic responses of emitting voltage spikes were reproduced in simulations of experimental devices. Other advantages including low power consumption and less heat generation, as well as relative insensibility to electric field noise, can also be expected from nanoionics circuitry.A simple answer to “Why nanoionics?” is that Nature chooses ions to sustain vital functions of the human body. On the bottom level of biological intelligence, different types of ion channels form the molecular basis for electrical activities in neurons and support the transmission and processing of information. While Nature has always served as a great inspiration to develop methods for achieving lower‐power and real‐time artificial intelligence systems, it is obvious that nanoionics holds great potential in the development of ionic information processors Figure 10. Desp By decreasing the size to nanoscale, a high density of interfaces (including grain and phase boundaries as well as free surfaces), which are structurally different from bulk, are obtained in nanostructured materials and can improve the crucial electrical performance of batteries. For example, nanocrystalline electrodes exhibit both high theoretical capacity and high Coulombic efficiency. The development of liquid nanoionics for energy generation seems slightly slow because of its low efficiency, which is at present approximately a few percents, compared with efficiencies of up to ≈95% for standard rotational electromagnetic generators. Whether the well‐developed reverse electrodialysis (RED)  or recently developed nanofluidic RED, their energy density and efficiency are not ideal enough for large‐scale commercial applications due to the instinctive drawback of slow ion‐transport speed. Increasingly more strategies are proposed to increase efficiency and implement parallelization in recent years. One way to achieve both purpose is to improve ion‐transport speed and efficiency by integrating multiple external energy sources, e.g., light or thermal energy and concentration gradient. Another way is to construct a series or parallel connected circuit; for example, Schroeder et al. reported a nanoionic energy system that can generate a high voltage of 110 V at open circuit by connecting thousands of ionic conductive gel compartments in series. Therefore, it is expected that all electron‐transport‐based energy and information processors can be soon redefined by nanoionics.Regarding ionic energy, solid‐state nanoionics has shown its broad applications, e.g., in solid electrolyte batteries.147 BySince nanoionics is an up‐and‐coming research field, it has been foreseen that many difficulties must be solved before its real translation. For nanoionics systems, nanoscale structure and ion‐transport properties are two important factors. What's more complicated is how to integrate these two factors into one system. Therefore, finding candidate materials that are suitable for nanofabrication will be the first challenge. Regarding solid‐state ion‐transport materials, construction of the nanostructure is required. Furthermore, it is difficult to process liquid or semisolid systems, e.g., ionic hydrogels, at nanoscale. Although some ionic interfaces or ionic cables have been reported to communicate with biological systems, ions used in such implementations are not as controllable as their biological counterparts without nanostructures. Random ion transport can only be ordered by an external electric field. In this case, “armor” is needed, e.g., 1D nanochannels and nanotubes, 2D layered space, or 3D porous structure, which can work as a template for these liquid systems or semisolid materials, thus enabling ion transport with precise controllability.Overall, nanoionics refers to a new area of research in which ion transport is conducted on a nanoconfined space. It may lead to innovative technologies one day by utilizing ions as conductive carriers. Such new methods and technologies could mimic many biological functions, even if they do not have the exact same mechanisms of biological intelligence. This, in turn, may facilitate manufacturing of nanoionic devices for use in science and even in daily life in the future, which may have significant advantages over some of today's electronic technologies. An evident example is the brain, which is a natural ionic system that still exceeds the capabilities of electronic processors in many tasks. However, we must admit that nanoionic devices are not expected to be faster than electronic ones owing to their slow ion‐transport speed. However, they may be able to carry out high‐level information processing such as writing and erasing information under the condition of low energy consumption. One of their potential applications is to construct nanoionics‐based computer memory devices that are much longer‐lasting than the electronic ones in use today. Such an application will be essential for safe, long‐term storage of information. Meanwhile, similar to the application in information and sensory technology, it is expected that nanoionics will play an important a role in energy conversion and storage, e.g., in supercapacitors.Nanoionics research is in its infancy and its potential is unlimited. Undoubtedly, many challenges have yet to be overcome. Nevertheless, given the rapid technological advancement in nanoscience, as well as the exceptional potential benefits, nanoionics research deserves exploration in the near future. Exploiting the full power of nanoionics to fabricate active devices that are currently dominated by nanoelectronics will lead to the invention of new artificial intelligence forms that are crucially needed and have the potential to be world changing.The authors declare no conflict of interest.