These topics were found to be comprehensible and coherent by both retail experts and consumers. The topics indicated that human concepts are primarily organized around goals and interactions , rather than their intrinsic features (e.g. defining a tomato by the fact that it has seeds and is fleshy). These results are consistent with the notion that human conceptual knowledge is tailored to support action. Individual differences in the topics sampled predicted basic demographic characteristics. Our findings suggest that human activity patterns can reveal conceptual organization and may give rise to it.Computational models using text corpora have proved useful in understanding the nature of language and human concepts. One appeal of this work is that text, such as from newspaper articles, should reflect human behaviour and conceptual organization outside the laboratory. However, texts do not directly reflect human activity, but instead serve a communicative function and are highly curated or edited to suit an audience. Here, we apply methods devised for text to a data source that directly reflects thousands of individuals’ activity patterns. Using product co-occurrence data from nearly 1.3-m supermarket shopping baskets, we trained a topic model to learn 25 high-level concepts (or The online version of this article (10.1007/s42113-019-00064-9) contains supplementary material, which is available to authorized users. Our everyday experiences shape the way we conceptualize and act in the world. Following this intuition, previous work using text corpora has proved useful in understanding the nature of language and human concepts , rather than their internal features (e.g. defining a tomato by the fact that it has seeds and is fleshy).intrinsic and extrinsic features in concept representation. One way that people may reason about categories is to decompose them into intrinsic features or parts  to learn lower dimensional, vector representations of word concepts , is a probabilistic approach in which documents are generated according to a mixture of probabilities over latent themes or topics . More generally, most spoken language is concerned with effective communication of relevant information . They may also ‘stem’ words to remove prefixes and suffixes of words that have similar semantic meaning (e.g. eat vs. eating). Moreover, the order of words in sentences can also make a big difference to sentence meaning (e.g. ‘dog bites man’ vs. ‘man bites dog’). However, most standard implementations of topic models . Overall, these results suggest that—contrary to early research on cross-classification of food  are not given separate codes however.Note that—unlike traditional uses of LDA in NLP—we did not remove commonly occurring items from documents (i.e. ‘stop words’). Whilst natural language may contain ‘stop words’ (i.e. common words with little semantic meaning such as ‘the’), we did not believe grocery transactions to suffer from the same problem. In the retail case, purchasing popular products, such as milk, bananas and bread, may be informative, perhaps indicating that the consumer is stocking essential items. The basket data was fully anonymised for general research purposes so as to not be personally identifiable.k) and in each case monitored the training and test log-perplexity to ensure model convergence and generalization, respectively (see In our experiments we applied Latent Dirichlet Allocation (LDA) to the data, using the machine learning library in Spark 1.6.0  had 25 topics. Models were trained for a maximum of 500 epochs, used the Online Variational Bayes optimization algorithm with an Stir Fry) to general in scope (e.g. Cooking from scratch). This therefore provides early support for the hypothesis that consumers primarily recruit thematic representations when conducting their grocery shop.The topics recovered by LDA were coherent and readily labelled by the authors. Table Christmas topic and chicken korma was eighth most relevant within the food for now topic.When proposing labels for the topics, the authors had access to the full item-topic relevancy matrix. In some cases, products outside of the top five were instrumental in determining the topic label. For example, mince pies (a popular dessert consumed during Christmas in the UK) were the seventh most relevant item within the To evaluate the appropriateness of the topic labels, we conducted a more detailed study. Specifically, a group of industry experts were asked to look through a sample of highly ranking products from within 10 randomly selected topics and confirm that the proposed labels were indeed representative of the grouped products.2www.dunnhumby.com), a customer marketing company with over 29 years of experience working with grocery retailers and fast moving consumer goods (FMCG) brands. Employees were asked to participate via the company intranet and were not remunerated. Fifty-one participated in the study. Participants had a wide range of roles within the business, including data analysts, category experts, company directors and client leads. Of these, 56.86% were male. Participants were surveyed in early December 2016 and were blind to the purposes of this study. The Ethics Committee at the UCL Experimental Psychology department approved the methodology and all participants consented to participation through an online consent form at the beginning of the survey.Participants were recruited internally within the UK headquarters of dunnhumby  and accompanying product descriptions from a single topic. Images were 540 × 540 pixels each. Descriptions appeared below each image in size 12 font. The displayed products were the 10 with the highest All participants were asked to label the same 10 topics in a random order. The dependent measure was the proportion of times that participants selected the topic label originally proposed during the model development phase. This proportion was then compared against a random baseline, to check whether participants were responding non-randomly.It was not feasible to survey participants about all 25 topics in the final LDA solution given constraints on employee time. Therefore, 10 topics were chosen from the original 25 to include in the survey.Participants were first briefed about the purpose of the experiment. After agreeing, they were then asked to label a group of products for 10 separate topics. Four possible labels were suggested using radio buttons. The order of the presented topics was randomized. One of the four labels was the ‘target’ label proposed by the authors whereas the other three were randomly selected from the remaining nine topics. After selecting a topic label, participants then confirmed their choice with a ‘Continue’ button, before seeing the next set of products from a randomly selected remaining topic. At the end of the study, participants were debriefed.SE= 0.015) of the 51 retail industry experts selected the same topic label as was originally proposed by the authors. A two-sided binomial test showed this to be significantly above chance (p < .001). Figure When asked to select the appropriate topic label for a group of products from a list of four possible labels, 92.8%  and 4 ‘staple-food’ topics that we believed unlikely to vary as much over the year .The results of the expert study discussed in the “To evaluate seasonal trends in topic prevalence, the same data used in the “To calculate the monthly prevalence of each topic, we hard-assigned each basket to belong to one topic, using the maximum topic probability. We then calculated an index indicating the relative popularity of a topic in a given month by calculating the proportion of baskets belonging to a given topic in a month and dividing it by the average topic probability for a given month across all topics.summer fruits and summer salad peaked in popularity during the summer months. Contrasting, baskets labelled with the Christmas topic peaked in popularity during December and the surrounding winter months. Low calorie options appeared to peak in January and steadily decline to its lowest level of popularity in December. The ‘staple’ topics shown in Fig. Figure Results from the survey of retail experts in the “” sectionTo understand whether the product representations identified by the topic model were meaningful to real consumers, we conducted a controlled experiment. Specifically, a large sample of British supermarket shoppers were shown a set of products; 5 of which were highly ranked from within a topic and one that was an ‘intruder’ product, randomly selected from one of the other topics. Participants were asked to select the intruding product. This paradigm is often used when evaluating the fit of computational models to human semantic memory . Participants completed the survey as part of a larger, monthly survey for 50 card loyalty points. The final sample consisted of 3840 participants, of which 59.47% were female. The modal age group was 55-59 (n = 501) and 724 participants did not disclose their age. All participants were from England, Scotland or Wales with the majority of respondents based in central England (n = 946). Participants were surveyed during March 2017. The Ethics Committee at the UCL Experimental Psychology department approved the methodology and all participants consented to participation through an online consent form at the beginning of the survey.Participants were recruited using dunnhumby’s consumer survey panel; Shopper Thoughts  alongside product descriptions. Five were the most relevant from within a topic and the other ‘intruder’ was the most relevant product from a randomly selected alternative topic.The dependent variable was the proportion of times that participants identified the intruder product. This proportion was then compared against a random baseline to assess whether participants were able to identify intruders significantly above chance levels.Participants each completed one trial in which they were asked to identify the intruding product. Participants were randomly selected to see one of the 10 topics also used in the retail experts study. This ensured that comparisons between the two related studies were consistent.SE = 0.007) were able to correctly identify the intruder product. A two-sided binomial test showed this to be significantly above chance (p < .001). Figure Of the 3840 British consumers surveyed, 74.1% (afternoon tea. Participants were most likely (51.7% of the time) to incorrectly suggest that ‘mint humbugs’ was the intruder. One possibility for this poor classification accuracy is that participants did not have enough context to interpret them correctly. In the afternoon tea topic, the top 5 items were predominantly fresh and ‘staple’ foods . Thus, seeing a packet of sweets (i.e. ‘humbugs’) among this fresh food may have appeared unusual. An analogous issue arises with the low maintenance cooking topic. Each topic is a probability distribution over thousands of products, so perhaps it is not surprising that a small sample of products could be ambiguous.One topic stands out for its below chance level of performance, afternoon tea topic may have been driven by the fact that most British people no longer regularly engage in this ritualistic activity. If experience shapes people’s mental concepts, then we would expect representations of certain products to vary between demographics. Supporting this view, consumers from Northern Ireland had an average probability for the Northern Ireland topic 7.5 × higher than the average across all regions. The fact that the model was able to recover such strong regional differences in consumers suggests that it should be sensitive to other individual differences in people’s experience of the world.Another possibility that is more core to our theory is that individual differences in experience may help explain some of these confusions. For example, the poor classification of the If the topic model proposed in this paper is representative of people’s semantic categories, then it should also be able to uncover individual differences in their representations. To test this assertion, we used logistic regression to predict  self-reported ageThe feature set for the supervised models comprised of the training set topic probabilities output by LDA, averaged at the customer level. These features were then used to predict customers’ self-reported age , region ) and gender. These self-reported values had been gathered by the marketing panel described in the “λ values of 0.1 to 1.0. Model selection was performed using the average predictive performance over 5 cross-validation folds. Baselines were calculated by predicting the majority class in each fold.To find the best performing model, we performed a grid-search between The age model was evaluated according to the average classification accuracy across the four classes. The gender and region models were binary classification problems, and thus evaluated in terms of the area under the ROC curve (AUC).Results showed that the models were able to predict age with an accuracy of 48.51%, region with an accuracy of 58.34% and gender with an accuracy of 57.17%, considerably higher than the guessing baselines of 36.85%, 50.00% and 50.00%, respectively.Rather than being solely defined by intrinsic features Plato , conceptChristmas topic in December and the Summer salad topic in the Summer.We tested this possibility by considering the shopping patterns of thousands of UK consumers. Using LDA, we found that the pattern of consumer purchases was highly revealing of people’s conceptual organization of these products. Topics ranged from specific and goal-driven (e.g. ingredients for a stir-fry) to very general (e.g. cooking from scratch). Interestingly, the topics tended to be goal-directed and situational, which is consistent with the notion that much of human conceptual knowledge is defined relationally and tailored to support action . One avenue for future research is to develop, apply, and evaluate topic models in which individuals organize into higher-level groups that can vary in terms of topic prevalence or even topic composition.If concepts gain meaning through the actions we take, then individual differences in experiences should be reflected in differences in conceptual organization. In support of this conjecture, topic prevalence varied across geographic regions. In our study of everyday consumers, poor performance for the topic Taxonomic and thematic cross-classifications of food are typically measured in free-sorting tasks, where participants must sort food items into groups Below is the link to the electronic supplementary material.


Micro- and nanofibers are historically-known materials that are continuously reinvented due to their valuable properties. They display promise for applications in many fields, from tissue engineering to catalysis or sensors. In the first application, micro- and nanofibers are mainly produced from a limited library of biomaterials with properties that need alteration before use. Post-modification is a very effective method for attaining on-demand features and functions of nonwovens. This review summarizes and presents methods of functionalization of nonwovens produced by electrostatic means. The reviewed modifications are grouped into physical methods, chemical modification, and mixed methods. Electrospinning  is one of the most effective methods of nanomaterial production. It has a very high throughput, enabling the construction of materials of different types of polymers. Fragile polymers, drugs, or even living cells can be electrospun. The physical conditions of electrospinning have been established . The tecThis review is based on the author’s 15 years of laboratory experience with on-demand electrospun nanofibers. As the main subject of the author’s works has been applications of nanofibers in biology and medicine, this review will mainly (but not only) deal with different types of scaffolds for tissue engineering, with some references to ceramic nanofibers. A vast amount of research represents nanofiber post-processing, so the author needed specified keys to select the presented material. The review is based on scientific recognition. For clarity, specific techniques are usually represented by two examples. The author omitted complicated, multiple-step post-modifications. The author have designed this review for newcomers in the electrospinning community, in order to give them well-established knowledge on the post-processing of electrospun micro- and nanofibers.The chapters start with the simplest physical modification methods, followed by more complicated chemical modifications, and end with the most complicated physico-chemical modifications .Physical surface modifications of electrospun nanofibers are directed towards thermally altering properties, stretching to change mechanical properties, the leaching or absorption of water-soluble polymers, plasma processing to increase the surface hydrophilicity, ultraviolet and laser ablation to produce patterned change of properties, and ultrasonic treatment directed towards increasing the porosity or fiber fragmentation  Table 1Table 1.β -lactoglobulin (BLG) and used heat to render fibers insoluble in water. The fiber-forming PEO used was as low as 25%, and the mat was heated to 100 °C to make it insoluble in water. The mat swelled in water and retained a fibrous structure, which suggested its possible use in regenerative medicine. Rhodamine B (RhB), as a model flavonoid, was incorporated into the mat and subsequently released. Min et al. [One of the simplest physical methods that affect the properties of electrospun nanofibers is temperature curing. Kowalczyk et al.  cured bon et al.  electrosn et al.  studied The thermal treatment of nonwovens and thermal matrix carbonization or degradation to form carbon or ceramic nanofibers is described in the chapter “Physico-chemical modifications.”Usually, as spun nanofibers have a low degree of molecular orientation, stretching can improve their crystallinity and strength. A generally higher degree of crystallinity accompanies a far lower elasticity and decreased rates of bio-degradation. The stretching of electrospun nanofibers is quite an unconventional method of modification. The strength and modulus results of single fibers are improved by up to two orders of magnitude when compared with nonwoven mats  Figure .co-caprolactone) (PLC) nonwoven used for tissue engineering. It suggested that pre-implantation stretching can drastically change the mechanical properties. Such treatment can improve tissue to implant matching. Zong et al. [co-glycolide) (PLGA) or poly(L-lactide) (PLLA) nanofibers led to anisotropic scaffolds used for cardiac cell cultures. Primary cardiomyocytes on poly(L-lactide) nanofibers developed functional contractile machinery (sarcomeres). They were electrically active.Lai et al.  producedg et al.  uniaxialg), e.g., poly(L-lactide-co-caprolactone) (PLC). PLC can be spun above Tg, but for electrospinning at room temperature, additional thermal treatment might be necessary.Thermal treatment and nanofiber mat relaxation may be essential for preventing the nonwoven mat from shrinking after its withdrawal from a target. The treatment is necessary for polymers electrospun below the glass transition temperature  or poly, are commonly used to improve the spinnability. They are also used as a fiber-forming polymer by mixing with non-spinnable polymers. These synthetic polymers, as well as natural ones, can be leached out of composite nanofibers to create nanoporosity. Poly (PVA) is highly hydrophilic, and draining it out of the polymer matrix may be challenging.Zhang et al.  studied A protein coating on scaffolds can increase the capacity of the cells to recognize the surface. Absorbed proteins can play a role as amphiphilic substances temporarily increasing the (bio) availability of the scaffold surface. It is one of the simplest methods available, needs no medical approval for use, and has no adverse effect on the mechanical properties of a polymer matrix. The technique is, therefore, especially valuable, even if the results are not as good as those obtained for permanently modified scaffolds.co-glycolide) as scaffolds for guided bone regeneration. They coated fibers by soaking them in poly(dopamine). Bone-forming protein −1 (BFP1) was introduced to nonwovens pre-coated with poly(dopamine) (PD) by incubating them overnight at 37 °C. Human mesenchymal stem cells seeded on the scaffolds formed an implant for the mouse calvarial defect model. The implants showed 2–3 times more bone regeneration after two months in vivo when compared with a non-implant group. A very appealing approach was presented by Zhang et al. [E. coli and S. aureus.Zhang et al.  soaked pg et al. , who covg et al.  covered 2), power, and treatment time. Plasma-treated polymers required further processing. The surface effects of plasma modification are only temporary and are gradually lost over time.Plasma treatment is a very convenient method, and is mainly used to increase the surface hydrophilicity. It is widely used in industry to modify polymer films . The mai4 or oxygen plasma to produce scaffolds for skin regeneration. Methane plasma modification slightly influenced the hydrophilicity compared to oxygen plasma. The latter caused a large increase in the wettability and cellular activity of human epidermal keratinocytes and fibroblasts [co-glycolide). The adhesion and proliferation of mouse fibroblasts were also positively affected. The degradation, in contrast, sped up, making the material less favorable for the production of scaffolds for tissue engineering [2 + H2, NH3 + O2, and Ar + O2, increased the hydrophilicity of nonwovens. Mouse fibroblasts seeded on plasma-modified poly(caprolactone) nanofibers gave higher proliferation and adhesion rates compared to untreated fibers. Plasma modification only had a limited negative impact on the material tensile properties [Poly(caprolactone) nanofiber meshes have been modified by radio-frequency (RF) plasma to enhance cell adhesion, differentiation, and proliferation. For fibroblasts, osteoblasts, and chondrocytes, oxygen plasma treatment was the most successful approach. It showed the greatest enhancement of the contact angle and material–cell interactions. The authors also noted that the surface roughness is a key factor in material–cell interactions . Electroroblasts . Ammoniaineering . A shortineering . Glow dioperties . De Valeoperties  very eleUV or laser beams can be applied for the patterning and controllable surface modification of electrospun nanofibers. These methods of selectively changing the surface properties can create a pattern of “well-like” 2D environments for directed cell culturing.co-glycolide) (PDLG) and poly(L-lactide-co-caprolactone) (PLC) nanofibers. The authors used a commercial germicide sterilization UV lamp (wavelength 254 nm). One hour of irradiation led to a ca. 40% decrease in the molecular weight and ca. 30% decrease in the tensile strength. Masked UV irradiation produced patterned nonwovens. Smooth muscle cells migrated to irradiated wells. Lim et al. [Yixiang et al.  studied m et al.  used a fm et al.  showed aUltrasonic treatment leads to the degradation of polymer chains. Its use in polymer nanofiber treatment is therefore limited to surface treatment and fiber scission. Scaffolds with pores enhanced by sonification have a cotton candy structure of lower mechanical properties; the polymer matrix is also pre-degraded.Lee et al.  used ultChemical modifications of nonwovens are far more complicated than physical ones. The main applications are stable surface modification, the crosslinking of water-soluble nonwovens, and precipitating mineral compounds on the surface. The main drawbacks are the need for the use of non-medically approved chemicals and the speeding up of polymer matrix degradation .Wang et al.  producedSurface hydrolysis is a standard procedure used to increase the hydrophilicity of polymer scaffolds. It helps cell attachment, proliferation, and differentiation. The process is pronounced not only on the surface, but also in the bulk polymer. It speeds up the degradation of polymer nanofibers.Boland et al.  used hydHydroxyapatite (HAP) deposition is an alternative method to direct addition of HAP nanoparticles to an electrospun solution. It creates biocompatible minerals on the surface of micro- and nanofibers, at places where cells will recognize them. This method is mainly used for bone tissue engineering.co-hydroxyvalerate) (PHBV) enclosed Bioglass. Materials were covered with HAP from SBF and displayed electrospun nanofiber degradation, which was the least extensive on PCL nanofibers (co-glycolide) nanofibers. Human osteosarcoma cell lines exhibited an excellent biocompatibility for the scaffolds. Cell adhesion, proliferation, and differentiation were reasonable for all mineral layers produced.Bretcanu et al.  used bionofibers . Chen etWater-soluble, bioactive polymers can be electrospun in a native bio-recognized form. The crosslinking of such nonwovens makes them insoluble and useful as scaffolds for tissue engineering.Chen et al.  crosslinPhysico-chemical modification is the most frequently used method of nonwoven post-processing. It allows the fabrication of nanomaterials of entirely new, non-spinnable materials (graphite nanofibers and ceramic nanofibers), surface grafting with polymers, and the stable anchoring of biological molecules on the surface of nanofibers  Table 3Table 3.3 bonds and ca. 10.5% of non-carbon atoms in the nanofibers. The micromechanical testing of numerous individual fibers gave an average modulus of 63 GPa and a diameter of 50–250 nm . Most frequently, metal oxide precursors do not form fibers. Spinnable polymers, such as poly(vinyl pyrrolidone) (PVP), poly, and poly(acrylonitrile), are added to form nanofibers. The polymer matrix is subsequently burned out in the oxygenated atmosphere, while metal oxide particles are sintered.Nanofibers of titanium dioxide nanoparticles suspended in poly(vinyl acetate) were deposited onto arrays of platinum electrodes, pressed to 120 °C, and calcinated at 450 °C. The material formed nanometer-size anatase crystals and showed an eight-fold increase in sensor resistance when used as a nitrogen dioxide sensor . TitaniuA comprehensive review of ceramic nanofiber synthesis and application is given by Panda .Plasma treatment was used to induce surface-functionalized groups. They were further used for the chemical binding of biomolecules or grafting with hydrophilic polymers by free-radical polymerization.co-glycolide) were subjected to oxygen plasma and in situ grafted with hydrophilic acrylic acid. Highly porous nonwovens with a surface-bonded poly(acrylic acid) moiety containing carboxylic groups had lower contact angles. Fibroblasts seeded on plasma-grafted nonwovens exhibited more significant attachment and proliferation when compared to unmodified nanofibers [Nanofibers produced from hydrophobic poly(glycolide), poly(L-lactide), and poly. The nonwoven was subjected to fast ammonolysis with 1.6-hexanediamine, followed by glutaraldehyde coupling and the covalent bonding of fibronectin to the spacer. The material strain decreased during the grafting, but the tensile strength remained unchanged. Porcine esophageal epithelial cells seeded on the modified scaffold displayed a proper phenotype and were much prominent when compared with the pristine one. Ghasemi-Mobarakeh et al. [co-glycolide)-poly(ethylene oxide)-NH2. The particular copolymer was terminated with the amino group used to couple the lysosome. The enzyme was covalently linked using ethylene glycol-bis (sulfosuccinimidylsuccinate). The proposed methodology can be used to fabricate nanofibers with other bioactive molecules attached to the surface.Zhu et al.  fabricath et al.  chemicalh et al.  used UV h et al.  electrosNumerous reputable review articles of nonwoven post-modification are available. They concern specific fields of the vast subject of electrospun nonwoven modification, including the surface functionalization of electrospun nanofibers for tissue engineering and drug delivery ; tissue As exemplified in Feynman’s famous statement—“there is plenty of the room at the bottom,”—there are a multitude of applications where nanofibers can be especially valuable. The technique was a curiosity when discovered; now, it is one of the most efficient and cheap ways to produce nanomaterials. The field of the use of such materials is far from being limited to the tissue engineering, wound dressing, sensors, and catalysis that the author has presented in this review. Being included in wearable electronics, power generation , telemedicine sensors, filtration and separation, health protection, and environmental remediation, the materials are also a part of the circular economy. The multitude of parameters governing electrospinning may seem scary, but they create unique possibilities to make any new material on demand. If the material is not good enough, one can use a multitude of surface modification techniques. Based on the steadily growing number of science articles that are discovering innovative materials and applications based on electrospinning and other ways of producing nanofibers , it is one of the new techniques that will change the world in the 21st century.


Agreement of molecular subtype did not significantly impact RFS and OS (p = 0.22 for both). The identified factors likely mirror intratumoral heterogeneity that might compromise obtaining a representative CNB. Our results challenge the robustness of a single CNB-driven measurement of Ki67-LI to identify luminal B breast cancer of low (G1) or intermediate (G2) grade.Reliable determination of Ki67 labeling index (Ki67-LI) on core needle biopsy (CNB) is essential for determining breast cancer molecular subtype for therapy planning. However, studies on agreement between molecular subtype and Ki67-LI between CNB and surgical resection (SR) specimens are conflicting. The present study analyzed the influence of clinicopathological and sampling-associated factors on agreement. Molecular subtype was determined visually by Ki67-LI in 484 pairs of CNB and SR specimens of invasive estrogen receptor (ER)–positive, human epidermal growth factor (HER2)–negative breast cancer. Luminal B disease was defined by Ki67-LI > 20% in SR. Correlation of molecular subtype agreement with age, menopausal status, CNB method, Breast Imaging Reporting and Data System imaging category, time between biopsies, type of surgery, and pathological tumor parameters was analyzed. Recurrence-free survival (RFS) and overall survival (OS) were analyzed using the Kaplan–Meier method. CNB had a sensitivity of 77.95% and a specificity of 80.97% for identifying luminal B tumors in CNB, compared with the final molecular subtype determination after surgery. The correlation of Ki67-LI between CNB and SR was moderate (ROC-AUC 0.8333). Specificity and sensitivity for CNB to correctly define molecular subtype of tumors according to SR were significantly associated with tumor grade, immunohistochemical progesterone receptor (PR) and p53 expression ( Reliable determination of molecular subtype is indispensable for prognostication and treatment decision in breast cancer (BC) , 2. EspeRecommendations to standardize Ki67-LI assessment recognized the need for further studies to evaluate comparability of Ki67-LI between CNB and whole tumor slides of surgical resection (SR) specimens . We thern = 484) with ER+/HER2− invasive BC at the Medical University of Vienna (MUW) were included in this retrospective analysis. Study procedures were approved by the Institutional Review Board of the MUW (1245/2017). Patients with primary operable luminal BC stages I–III who received diagnostic CNB and curative surgery without neoadjuvant therapy between 2010 and 2012 were eligible to participate. Age, menopausal status, type of surgery (breast conserving vs. mastectomy), and previous BC history were recorded for each patient. Clinical and pathological patient characteristics are described in Table Patients ( the MUW 245/2017.All patients underwent pre-operative either stereotactic vacuum-assisted (9G), ultrasound core needle 14G), or magnetic resonance imaging (MRI) vacuum-assisted (9G) guided breast biopsy according to the European Society of Breast Imaging (EUSOBI) guidelines . In some4G, or maWorkup was carried out according to the EU guidelines , 22 and Assessment of ER, progesterone receptor (PR), HER2, Ki67-LI, and p53 was performed using Ventana BenchMark Ultra  according to the ASCO/USCAP guidelines , 27. Ki6Continuous data are presented as median, minimum, and maximum due to skew distributions. Categorical data are presented as absolute frequencies and percentages. Sensitivities to correctly predict LumB molecular subtype in SR and specificities to predict LumA molecular subtype by CNB are given partly together with 95% confidence intervals according to the method of Wilson. To assess diagnostic ability of CNB to predict SR over several CNB cut-points, a receiver operating characteristic (ROC) curve and its area under the curve are given.Associations between two binary variables are tested by chi-square test or Fisher’s exact test in case of small cell numbers. To test ordinal variables between LumA and LumB molecular subtype, a trend chi-square test was used and an exact version was used in case of small numbers.Recurrence-free survival was defined as the interval between the CNB and the first evidence of relapse at any site or incidence or contralateral breast cancer. Overall survival was defined as the interval between CNB and death. Survival rates were estimated with the use of the Kaplan–Meier method.p values are two-sided and p ≤ 0.05 was considered significant. Calculations were performed by the statistical software SAS© .All Median Ki67-LI was 21.4% and 21.7% for CNB and SR, respectively. A substantial agreement of Ki67-LI between CNB and SR specimens was observed. The ROC for molecular subtype in CNB to correctly diagnose LumA in SR (Ki67-LI < 20%) resulted in an area under the curve (AUC) of 0.8333 Fig. , indicatAgreement of molecular subtype determination by Ki67-LI was observed in 386 (79.8%) patients. In 98 20.3%) patients, Ki67-LI in CNB and SR were discordant. Examples are shown in Fig. 0.3% patiIn our analysis, we identified tumor grade , lower PR expression, higher p53 expression, and LVI as tumor-related factors in CNB that significantly influenced agreement of molecular subtype. A dependence between ER expression in CNB and Ki67-LI for LumA and LumB tumors could not be demonstrated, mainly due to small group sizes with ER < 80% in CNB (7 LumA tumors and 14 LumB tumors). Results are shown in detail in Table In SR specimens, tumor grade , tumor size (pT), multifocality, lymph node involvement (pN), LVI, and performed mastectomy were identified as tumor-related factors that significantly influenced agreement of molecular subtype. Results are shown in detail in Table p = 0.22, respectively). However, patients with correctly diagnosed LumB tumors tended to have a worse RFS and OS, as shown in Fig. Survival data was available for 390 patients, of whom 177 (30%) were correctly diagnosed with luminal A tumors and 199 (51%) were correctly diagnosed with luminal B disease in CNB. In 43 patients (11.2%), a LumA tumor in CNB was upgraded to LumB by SR and in 31 patients (7.9%), a LumB tumor in CNB was downgraded to LumA by SR. Kaplan–Meier analyses revealed no significant differences regarding RFS and OS . By evaluating the influence of the time interval between CNB and surgery, we analyzed whether wound healing processes after CNB impact local tumor proliferation. Previous studies suggested that surgery time interval might influence Ki67-LI change after CNB, longer STI leading to a higher increase in Ki67-LI . In our In our study, further clinicopathological factors, most importantly G and size, PR and p53 expression influenced sensitivity and specificity of molecular subtype determination. These factors are likely indicators of intratumoral heterogeneity and might therefore predict reliability of molecular subtype assessment in CNB. Previous analyses confirmed that adverse clinicopathological factors such as high G, LVI, and high p53 expression were significantly associated with a high Shannon index regarding copy number variation of oncogenes, implicating these factors as indicators of intratumoral heterogeneity .ER-negative tumors display higher concordance rates between CNB and SR than ER-positive tumors . In our We now report that additional factors, such as multifocality, PR, and p53 expression, also affect sensitivity of Ki67-LI assessment. Considering these factors is likely to increase the acuity of molecular subtype determination.In our study, all separate grading components, namely glandular differentiation, nuclear pleomorphy, and mitotic count, inflicted a significant impact on correctly classifying luminal molecular subtype. Although applying histological grade and mitotic index might reliably identify LumA tumors, Ki67-LI is needed to correctly classify LumB tumors . In our At present, no uniform cutoff levels for Ki67-LI have been defined and discrepancies regarding thresholds and their implications for clinical decisions remain –19, 41. Our study revealed no statistically significant differences in RFS and OS depending on agreement of molecular subtype between CNB and SR, which might be explained by the limited cohort size and the low number of events.Previous studies suggested that digital image analysis vs. visual determination might improve molecular subtype determination by immunohistochemistry ; howeverIn our study, pathological factors such as tumor grade, size, PR, and p53 expression significantly influenced agreement of Ki67-LI and luminal molecular subtype between CNB and SR. These factors mirror tumor heterogeneity and can objectivize molecular subtype determination in CNB.More importantly, our results question the robustness of a single CNB-driven measurement of Ki67 in luminal BC of low (G1) or intermediate (G2) histological grade and warrant further investigations to improve the validity of molecular subtype determination in these cases.


This paper reviews the occurrence of antimicrobial resistance (AMR) in insects, rodents, and pets. Insects , rodents , and pets  act as reservoirs of AMR for first-line and last-resort antimicrobial agents. AMR proliferates in insects, rodents, and pets, and their skin and gut systems. Subsequently, insects, rodents, and pets act as vectors that disseminate AMR to humans via direct contact, human food contamination, and horizontal gene transfer. Thus, insects, rodents, and pets might act as sentinels or bioindicators of AMR. Human health risks are discussed, including those unique to low-income countries. Current evidence on human health risks is largely inferential and based on qualitative data, but comprehensive statistics based on quantitative microbial risk assessment (QMRA) are still lacking. Hence, tracing human health risks of AMR to insects, rodents, and pets, remains a challenge. To safeguard human health, mitigation measures are proposed, based on the one-health approach. Future research should include human health risk analysis using QMRA, and the application of in-silico techniques, genomics, network analysis, and ’big data’ analytical tools to understand the role of household insects, rodents, and pets in the persistence, circulation, and health risks of AMR. Enterococcus faecium, (2) Staphylococcus aureus, (3) Klebsiella pneumoniae, (4) Acinetobacter baumannii, (5) Pseudomonas aeruginosa, and (6) Enterobacter spp.) [Clostridium difficile, (2) Proteus spp., and (3) pathogenic Escherichia coli [Antimicrobial resistance (AMR) is an emerging global health concern to both animals and humans. In general, AMR is a collective term referring to the capacity of microorganisms (often pathogenic) to develop physical or biochemical mechanisms and processes that render antimicrobial agents ineffective, including antibiotics . Compareer spp.) . Recentlumannii,  Pseudomoosa, and  Enterobahia coli . Hence, The existing literature on AMR, including review papers, focused on the following aspects, among several others—(1) human infections, including nosocomial ones or those associated with healthcare facilities ,8, (2) oAn increasing body of recent evidence shows that antimicrobial resistance occurs in insects, wildlife, including rodents and birds, and even companion animals or pets ,18. SeveTherefore, the present review posits that (1) insects and rodents, including edible ones and those associated with household settings and pets, harbor and act as reservoirs of a diverse range of antimicrobial-resistant microorganisms and resistance genes of human health concern; (2) the animal compartments comprise insects, rodents, and pets, which act as transmission vectors and sentinels or bioindicators of AMR; and (3) the persistence and dissemination of AMR via insects, rodents, and companion, pose significant human exposure and health risks. To address these hypotheses, the source-pathway-receptor-impact framework was used to track the occurrence and circulation of AMR in the environment–animal –human interface.AMR is classified into two groups—intrinsic or acquired. While intrinsic AMR results from the ability of the microorganism to survive in the presence of an antimicrobial agent because of an inherent property of the microorganism, acquired AMR emanates either from gene mutation or by the microorganism acquiring extra gene coding for a resistance mechanism . AlthougAs antimicrobials are commonly used in the management of microbial infections in veterinary and human medicine, and also as growth promoters in livestock, ABR is exacerbated in natural and man-made environments and consequently attracts relatively wider research attention ,33,34. SThe interfaces among humans, animals, and the environment are hotspots for the exchange of antimicrobial-resistant microorganisms and their resistance genes . PreviouAntimicrobials are continuously discharged into the environment due to their intensive usage, resulting in the emergence of antimicrobial resistance ,43. MicrYersinia pestis, which causes plague, can be transferred from one rodent to another through flea bites [Apart from occurrence in livestock and wild animals via interactions through shared environments, AMR is detected in cockroaches ,45,46, wea bites . This caea bites . Anotherea bites .The main mechanisms for the development of antimicrobial resistance related to drug usage are (1) new opportunistic pathogenic microbes emerge, which are usually antimicrobial-resistant; (2) new acquired resistance mechanisms emerge; (3) gene mutation in the host chromosome or plasmid-borne occurs; and (4) due to the dispersion of old resistance genes into new host microorganisms . A more Acquired resistance is mainly transferred via HGT of environmental or microbial ARGs, and this is facilitated by mobile genetic elements (MGEs) . While iThe genetic modifications in acquired resistance alter the defenses of the microorganisms through several pathways. These include (1) altering the target receptor of the antimicrobial agent, (2) altering membrane permeability to antimicrobial agents, (3) enzymic inactivation of the antimicrobial agent, (4) active transportation of the antimicrobial agent via efflux pumps, or (5) by directing metabolic routes to avoid disruption . It is lFollowing the capture of the host insects or animals such as cockroaches and flies, the microorganisms can be isolated using classical methods  and then subjected to standard protocols for antimicrobial susceptibility testing ,55,56,57Cultural methods still dominate studies on AMR. There is a need to improve on methods and implore advanced technologies, such as the sequencing of the whole genome based on antimicrobial-resistant microbial isolates. This could be helpful in providing critical information, e.g., for identifying shared microbial strains, antibiotic resistance genes, and mobile genetic elements ,60. UsinTo quantify AMR, it is essential to understand the advantages and disadvantages of conventional culture-based methods and molecular-based methods. Culture-based methods require immediate sample treatment and are limited to particular microbial strains and bacteria that can be cultured on synthetic media . This apE. coli and Yersinia strains [In addition to qPCR, a range of modern techniques are used to detect and quantify ARB. For example, following an extraction, gel electrophoresis was used to determine the quality of the DNA . A simil strains . Yet ano strains . The app strains . Neverth strains . In addi strains ,65. The  strains ,65. TheyMusca domestica), false stable flies (Muscina stabulans), and stable flies  [Several insects, including those associated with household settings, are reported to harbor AMR . To datecitrans) . HoweverE. coli, exhibiting multi-drug resistance to several antibiotics, including cloxacillin, tetracycline, ampicillin, neomycin, and bacitracin [Insects such as houseflies and cockroaches can move unrestricted between different environmental compartments, allowing them to easily spread antimicrobial-resistant pathogens. This high dispersal ability of insects is set to promote the occurrence and persistence of antimicrobial resistance in humans, emanating in anthropogenic activities in other sectors such as agriculture. For instance, houseflies and cockroaches successfully transmitted antimicrobial-resistant bacteria obtained in pig manure from two commercial confined swine farms ,51. In acitracin . The stucitracin , the livThe alarming rise in antimicrobial resistance, even to the last-resort antibiotics, is an unprecedented challenge to public health, as it limits treatment options of microbial infections. Thus, the World Health Organization has called for a “One Health” approach to find effective solutions to address this challenge . While tP. aeruginosa could multiply in the gut of cockroaches and be excreted for up to 114 days. These findings show that, in addition to being AMR vectors, houseflies and cockroaches are also amplifiers of antimicrobial-resistant pathogens and their associated resistance genes. However, more work needs to be done to ascertain this phenomenon in detail and confirm its occurrence in other insects and rodents. Pathogens in the guts of houseflies and blow-flies were three times greater than on their body surfaces [Most insects prefer damp areas, and dead and decaying matter, including sewers, carcasses, human and animal fecal matter, and garbage, among others. Through frequenting such unsanitary environments, the insects come into contact with pathogens and subsequently transmit them to other compartments, including humans. The exoskeleton and alimentary canal (or gut) of the studied insects acted as reservoirs for antimicrobial-resistant microbiota, antimicrobial resistance genes, and mobile genetic elements. Besides the exoskeleton and feces, the gut, saliva, or vomit through regurgitation is another additional reservoir and source for transmission . When fesurfaces . These fsurfaces ,76. Thessurfaces . It shouCochliomyia macellaria , the red-tailed flesh fly, Sarcophaga haemorrhoidalis , and the black garbage fly, Ophyra leucostoma ) were sampled using the sweep nets, but the analysis of their guts revealed almost non-existence of the tested pathogens [In one study, several types of flies  per fly per mL.Insects are known to invade and inhabit human foods, whether accidentally or intentionally . Some ofE. coli and other Enterobacteriaceae [E. coli to Acinetobacter sp. and Pseudomonas fluorescence within the intestines of the flies. In a similar study, one-day-old houseflies were fed with an antibiotic-resistant E. coli suspension, followed up to the next-generation [E. faecalis [E. coli [Another study demonstrated that the gut of cockroaches represented a favorable environment for the horizontal transfer of resistance genes between eriaceae . The sameriaceae . The samneration . The resfaecalis  and ampi[E. coli .Blattella germanica) and the digestive tract of houseflies were dominated by the multi-drug-resistant bacteria, E. faecalis, along with the associated ARGs; tet(M), erm(B), gelE, esp, asa1, and the Tn916/1545 transposon family obtained from swine farm manure [Most insects captured and studied in most studies harbored antimicrobial-resistant microbiota, ARGs, and MGEs ,51,58. Tm manure . Metagenm manure . Hence, K. pneumoniae in neonates within the studied hospital environment. This conclusion was further strengthened when the number of new cases dropped significantly, following intervention measures that resulted in eliminating the insects. Similarly, a direct correlation was observed between the density of flies and the incidence of Shigella infections in Bangladesh [The antimicrobial-resistant microorganisms carried on insects can be transmitted to other insects and surfaces through different mechanisms. This transmission could have severe public health implications. For example, a study conducted in South Africa reported no differences in the bacteria isolated from cockroaches and patients in a hospital, during an outbreak . The autngladesh .E. coli carried on the outer surfaces of flies was transferred to different food types, including steak, milk, and potato salad [Salmonella reported the ARB were transferred to nymph grown under sterile conditions [Salmonella-fed cockroaches and the sterile ones harbored the test organism. This study demonstrated that cockroaches (and other insects) could pick up bacteria  from dirty environments and transfer them to clean environments and other insects. Allen [Mycobacterium tuberculosis. The feces from the insects were then collected and examined for the presence of the bacilli, and it was observed that the bacterium was present in the insect’s feces for up to eight weeks, at room temperatures. Using metagenomics, it was also demonstrated that the intestinal microbiome of cockroaches shared up to 90% similarity with that of the insects’ feces [On the one hand, the ARB occurring on the outside layers of the insects, usually on the legs and mouth, are transferred mechanically as the insects interact with different types of surfaces and other insects. The mechanical transfer was reported in a study that demonstrated that to salad . On the to salad ,96. In oto salad . It was nditions . It was s. Allen  demonstrs’ feces , furtherIn addition to sharing their habitats with humans, some insects serve as food, due to their high energy content. With a global shift to insects as alternative protein sources , humans Salmonella spp. to ascertain whether cockroaches could acquire ARB from contaminated sites and subsequently transmit these to clean surfaces, water, and other cockroaches [One laboratory study used nalidixic acid-resistant kroaches . The reskroaches , mosquitkroaches , and bedkroaches , the traEarlier literature suggests that effective management for the control of the spread of AMR through insects should target the reduction of insect populations . Given tDespite the numerous studies on the role of insects as reservoirs and vectors of AMR transmission, a number of gaps exist. Most studies were carried out on houseflies and cockroaches, and only a few studies looked at other insects such as spiders, butterflies, mosquitoes, moths, for example, with which humans regularly come into contact with in households, gardens, and recreational facilities like parks. This is particularly crucial for resource-limited settings in sub-Sahara African countries, where the tropical climate promotes the proliferation of insects, resulting in regular exposure to other insects, such as ants, beetles, grasshoppers, and spiders.Bacillus thuringiensis, carrying antimicrobial and insecticidal resistance genes [Studies on AMR in insects mostly focus on understanding the prevalence and potential transfer to humans and other animals. However, given that most resistant organisms present in insects are acquired from human-impacted environments, insects could be used as sentinels for studying the resistome circulating within a community. The control of insect populations is mostly carried out with insecticides that contain heavy metals, which co-select for AMR ,109,110.ce genes . Thus, tPrevious studies are mostly on antibiotic resistance associated with antibiotics and bacteria. Little information exists on the potential of insects to act as reservoirs of drug-resistant viruses and fungi. Another critical gap for establishing the risk associated with ARB and ARGs in household insects is the lack of information regarding the transfer efficiency of the insect bacteria to different environments . To obtain such information, it is necessary to understand the proportion of resistant bacteria within different insects. In addition, how long the insect would have to be in a contaminated environment to carry a substantial number of bacteria and how much contact time would be necessary to deposit substantial amounts of these bacteria to different surfaces, need to be investigated further.E. coli bacteria to Yersinia pestis, resulting in the antibiotic-resistant Y. pestis from patients in Madagascar [As insects are potential vectors of human diseases, the presence of pathogens in insects is of importance and should be explored further. Research shows with some certainty that insects can be a source of AMR . For insdagascar ,113.Muscidae and Calliphoridae can transmit microbial organisms, which besides bacteria also include fungi and even viruses [As mentioned earlier, houseflies belonging to the families  viruses ,117,118. viruses ,120,121. viruses . They us viruses . Their n viruses . Their m viruses ,124. Thi viruses ,90. In h viruses .Houseflies are suggested as good candidates to use as sentinels in the surveillance of prevalence and spatial distribution of AMR in microbiota, such as bacteria in humans and animals, for several reasons. Houseflies have unrestricted movement such that they commonly occur on or around humans and animals, are easy and cheap to catch using simple traps, and are not subject to ethical issues, as with people . These fStaphylococcus aureus isolated from cockroaches sampled from houses and restaurants was shown to exhibit AMR, and this poses a health risk as the antimicrobial-resistant S. aureus can potentially be easily transmitted to humans from the cockroaches [Just as with houseflies, cockroaches are also known to spread AMR microorganisms such as bacteria, viruses, helminths, and protozoan, normally through food contamination in house dwellings ,126,127.kroaches ,131,132.Edible insects, which form part of some human diets, also present a cause for concern when it comes to the spread and transmission of AMR in households. Although there is not much research done on AMR in this area, a few studies showed that some edible insects containing different microorganisms in their guts contribute to the distribution of ARGs ,135,136.There are increased concerns with the mass breeding and continued release of genetically modified (GM) insects, such as mosquitoes and agricultural pests. As these genetically modified insects are bred, the antibiotic tetracycline is normally added to their feed, and overtime, these can develop AMR in their microbiota . The maiOverall, in the case of AMR in insects, previous studies mostly focused on insects of economic interest such as agricultural insects, bees, and those insects normally associated with pathogens of medical interest such as flies and cockroaches. Insects that do not necessarily fit in these categories are somewhat overlooked, yet they might be significant reservoirs, vectors, and transmitters of AMR.Besides insects and parasites ,149, rodYersinia pestis, Salmonella Typhimurium serotype, Streptococcus moniliformis, E. coli, and Yersinia enterocolitica, some of which were observed to be resistant to antimicrobials and yet they are responsible for causing different types of diseases in humans [The significance of rodents, including household rodents, as a reservoir of AMR, is inadequately recognized, with barely any large-scale research on this, as compared, for example, to AMR in humans. However, these small mammals play a crucial role as sentinels, reservoirs, and vectors of AMR, especially where anthropogenic factors such as pollution are involved or in one way or the other they associate with human waste ,154,155.n humans ,162,163.n humans . Evidencn humans .Mus musculus) that flourish in urban areas including human dwellings, pose a huge health risk to other species including humans, as they contaminate the environment with pathogenic microbes and in the process vectoring and transmitting any form of AMR organisms that they carry. The house mouse, commonly found indoors where it associates with humans, was reported to carry some pathogenic organisms such as C. difficile [Campylobacter [Salmonella [In other examples, rodent species such the house mouse  [In the food chain, rodents are normally one of the initial links close to the bottom of the chain and therefore can easily become a microbiological transmitter for several predators, including humans. In several parts of the world, including Africa and Asia, wild rodents form part of the traditional household diet, and if such rodents are AMR contaminated and then ingested, this might easily become a pathway through which AMR-resistant organisms carried by these rodents can be transmitted to humans. Wild rodents such as riverbank voles and wood mice harbor antimicrobial-resistant bacteria  , and if Yestinia pestis in bubonic plague patients in Madagascar, which was linked to horizontal gene transfer from rat fleas [Indirectly, rodents and even pets might harbor parasites possessing antimicrobial resistance, which might be transferred to humans via such parasites. A typical example is the emergence of antibiotic-resistant at fleas . The rolThe capacity of rodents to act as reservoirs and vectors of AMR points to their potential use as sentinels of AMR occurrence, transmission, and potential human health risks. Sentinels or bioindicators are organisms with the potential to be used as an early warning system for human health risks. A few studies showed that small animals, including mice, voles, and insectivores, are effectively used as sentinels of antimicrobial-resistant microorganisms and their ARGs ,41. In tPets or companion animals are domesticated animals normally kept within or near a household for purposes of companionship. These include dogs, cats, rabbits, birds, horses, rats, rabbits, guinea pigs, fish, snakes, and amphibians. Other reasons for pet ownership include safety, exercise, and as service animals trained to carry out specific tasks for the benefit of an individual, with a physical, mental, or sensory disability. Incidentally, owning pets can also reduce alienation, particularly in contemporary high-tech urban societies, which resulted in reduced human physical interaction. Medical evidence shows that pets can improve coping with cardiovascular diseases, can reduce depression ,180,181,S. aureus compared to healthy animals [Klebsiella and Acinetobacter spp., is an emerging human health concern [Pets are susceptible to many diseases such as urinary tract infections, post-operative wound infections, dental infections, and respiratory tract infections that might require the use of antimicrobials ,188,189. animals . Resista animals , recent  animals . Moreove concern ,200,201.Staphylococcus spp. from ear canals (otitis externa) of pet dogs were resistant to at least one antibiotic, while 77.1% of Staphylococcus isolates were multi-drug-resistant [E. coli isolates from UTI were most resistant to oxytetracycline and ampicillin [P. aeruginosa and E. faecalis, over ten years (1999–2009) [In one study, all isolates of esistant . The leaesistant ,210. E. picillin . In anot99–2009) .Pasteurella, Salmonella, Brucella, Yersinia enterocolitica, Campylobacter, Capnocytophaga, Bordetella bronchiseptica, Coxiella burnetii, Leptospira, Staphylococcus intermedius, and Methicillin-resistant S. aureus [Enterococcus spp.). The possibility of resistant bacteria passing resistance genes to microbial flora endemic to humans has long been a concern [Enterococcus spp. are considered endemic bacteria to the gut of humans and pets, they can act as reservoirs of AMR genes that can be transmitted to pathogenic bacteria such as S. aureus and Streptococcus spp. [E. coli in dogs and cats [Companion animals can transmit AMR bacteria to humans through direct contact or indirectly in their shared environment. Pets transmit several bacterial infections to humans via saliva, urine, feces, aerosols, and are major sources of zoonotic diseases. Some of the common pathogens causing bacterial infections from dogs include . aureus . AMR was concern . Althougcus spp. . Howevercus spp. ,216,217,and cats . This seDue to their closeness and direct contact with humans, a high risk exists for the transmission of AMR from pets to humans (and vice-versa). Human–pet interactions also occur via saliva, urine, feces, aerosols, and direct dermal contact, further increasing the risk of AMR transmission. Dogs and cats are among the most studied pets, but there is a need to understand the occurrence and potential transfer of AMR to humans via other under-studied pet animals .The identification and estimation of the adverse human health effects of AMR, including disease incidences and outbreaks, are challenging . This isTo date, most review studies on general antimicrobial-resistant microorganisms and exposure to humans overlooked the role played by flies, rodents, or household animals or pets in transmission and risk to human health ,225,226.Yersinia pestis isolated from patients in Madagascar was a result of the transmission of these bacteria through infected fleas to humans. It was inferred that there was HGT of AMR from E. coli bacteria to Yersinia pestis within the gut of fleas, which in turn then transmitted the antibiotic-resistant Y. pestis to humans [In one study, it was suspected that an antibiotic-resistant o humans ,113. In o humans . In anoto humans ,66. ThesTribolium castaneum, Herbst, and Tribolium confusum, Jacquelin duVal, respectively) had antibiotic-resistant and low concentrations of virulent enterococci in their guts and were shown to harbor MGEs, Tn916/1545 family of conjugative transposons that commonly host the ermB and tetM genes [E. faecalis OG1RF—pCF10 strain with antibiotic resistance from contaminated poultry and cattle feed to sterile feed [Enterobacteriaceae resistant to extended-spectrum cephalosporins were detected in dogs from households whose owners had a rectal culture positive for extended-spectrum cephalosporin-resistant Enterobacteriaceae [It can be inferred that the insects are also capable of transferring antimicrobial-resistant pathogens, their genes, and MGEs, if present, to humans. This is because some of the identified pathogens in the above studies harbored AMR traits to several antimicrobials ,227. FortM genes . In contile feed . In one eriaceae . ComprehDespite the lack of strong evidence, the emergence and spread of AMR pose potential human health risks. Multi-drug-resistant human pathogens are one of the major causes of morbidity and mortality in patients undergoing some challenging surgeries, such as hip replacements and organ transplants ,230. AMRSalmonella strain was isolated from two rodent meat and food products (Rattus tanezumi and Bandicota indica) from Thailand markets [Salmonella was acquired from the live rodents or handling and processing of the rodent meat [The occurrence of AMR in rodents and insects, including edible ones, and pets, raised human health concerns. Yet, evidence directly linking such AMR to human exposure and health risks remains weak. Here, selected studies pointing to potential human health risks are summarized. For example, the antimicrobial-resistant  markets . Howeverent meat . Evidencent meat . In anotent meat . Similarent meat .Most edible rodents and insects reported in the literature are also widely consumed in Africa. However, similar to other regions, the link with human health risks is still poorly understood, highlighting the need for further research. The consumption of edible rodents and insects is putatively high in developing regions , especially among low-income households who cannot afford meat. Therefore, these regions might represent ideal sites to understand the potential human exposure and health risks associated with the consumption of edible rodents and insects with AMR. Therefore, comprehensive studies are required to understand the occurrence of AMR in raw or uncooked rodents and insects and the impacts of handling and culinary practices on AMR at the point of consumption.The lack of compelling evidence on the human exposure and health risks associated with AMR in rodents, insects, and pets might reflect the methodological limitations of existing studies predominantly focusing on the occurrence and characterization of AMR. In these studies, the mere occurrence of AMR of human health concern is often interpreted as posing human exposure and health risks. However, estimating human exposure and health risks is a non-trial task for several reasons. First, there is a need for the accurate quantitative determination of the likelihood or probability of the transfer of antimicrobial-resistant organisms or resistance genes, and mobilome from the rodents, insects, and pet reservoirs and vectors to intermediate vectors or humans. Such transfer might occur via HGT facilitated by MGEs or via other human intake pathways, such as dermal contact, ingestion, and inhalation. Second, there is a need to estimate the frequency and magnitude of human exposure to AMR in such vectors. It is important to note that not all antimicrobial-resistant microorganisms and their resistance genes constitute human pathogens. The antecedent human health conditions  and age might affect human susceptibility, further complicating the estimation of human health risks.drought on fertile grounds’.Insects, rodents, and to some extent, pets are an integral component of societies in the developing regions of Asia, South America, and Africa ,235. ForQuantitative microbial risk assessment (QMRA) provides an ideal tool to overcome the methodological limitations of current studies, while simultaneously enabling a quantitative determination of human exposure and health risks of AMR in rodents, insects, and pets. QMRA is a systematic approach for identifying, evaluating, and estimating the human exposure routes and potential health risks of AMR, based on the source–exposure pathway–receptor–impact framework . QMRA usConsidering the risks arising from the spread of AMR, as outlined in the previous sections, there is a need to continuously assess the associated human health risks. The guidelines and recommendations related to the general assessment of AMR risks are outlined elsewhere ,244. Thumonitoring the usage of antimicrobial in companion animals,the extent to which the AMR occurs in pets, insects, and rodents entering households unintentionally,the association between AMR in rodents, insects, and animals of interest, and humans coming in contact with such animals, andthe routes via which AMR microorganisms and/or ARGs can be transmitted between household animals and humans.The risk assessment of human health threats should predominantly be based on data on the following:Despite the knowledge on the role of livestock animals in the spread of AMR, companion animals, such as cats and dogs, were for long not regarded as a significant transmission of ARGs or antimicrobial-resistant microorganisms to humans. Therefore, data on human health risks traceable to AMR in insects, rodents, and pets are scarce. Hence, further research efforts are required to generate information important for human health risk assessment in this context. The limited understanding of the occurrence of AMR in companion animals and other animals kept as pets in household settings (including rodents), coupled with the poorly understood risk factors, and transmission routes make it difficult to conduct comprehensive quantitative risk assessments. This position was also highlighted by the European Medicines Agency . SimilarThe assessment of antimicrobial usage and AMR in cats and dogs was recently performed and reported for Belgium, Italy, and The Netherlands . DespiteConsidering the possible transmission of AMR from companion animals to humans, as reported in China in relation to colistin , it is iStudies on quantitative health risk assessment should also cover the identification of routes of AMR transmission from pets to owners, as this is pivotal to develop management strategies and guidelines. This is, however, a particularly challenging task that should be coupled with studies addressing the frequency of AMR in animals and humans. It is also further complicated by the fact that there can be a bilateral transmission of AMR between animal and owner, making the assessment of the initial source of these transmissions even more challenging. One approach is to gather detailed information on the history of antimicrobial use in pet owners and pets , level of household hygiene, type, and frequency of intimate contacts between owner and pet, and the lifestyle of pets . One should also note that transmission events might not only occur during the use of antimicrobial drugs in animals, but also after the microorganisms with AMR colonize the pet, following their excretion and contamination of households .Since it is virtually impossible to conduct experimental studies directly addressing the transmission of AMR from actual pets to their owners, some more definitive conclusions could be drawn from prospective studies. Such research should assess the baseline level of AMR in owner and companion animals and follow it consecutively, along with tracking the history of antimicrobial use in both compartments. To date, no such investigations were conducted that highlight the urgent need for such studies. The application of a similar assessment of human health risks related to rodents and insects that are not intentionally kept in household settings but occur temporarily, is more challenging. This is due to difficulties in collecting the animals for material sampling, although one should note that such investigations were conducted on small mammals  outside household settings .The results indicate that rodents and insects can be sentinels or bioindicators for AMR transmission in the environment . Other sOne should note that AMR is, as stated by the WHO , “a compMitigation measures include (1) national and international guidance and monitoring on the use of antibiotics to limit the use of CIAs and non-essential use; (2) non-clinical interventions for animal health such as good feed, pet hygiene, regular check-ups, vaccination, and control of pests to improve animal health while reducing the use of antibiotics; (3) training of veterinarians on the selection of appropriate therapy to reduce antimicrobial resistance; (4) education of owners about good hygiene practices ; (5) raising awareness among owners to strictly adhere to recommendations on the use of antimicrobial use ; and (6) antimicrobial susceptibility testing for empirical guidance on the choice of antibiotics. For example, soft diets in dogs and cats are associated with high incidences of dental diseases, and harder, chewy foods requiring vigorous prehension and mastication improved dental health .limiting the use of antimicrobials in pets only to the clinically justified applicationsraising the awareness of the complex AMR issue within the veterinary specialists and even among the pet ownerslimiting direct contact between pet owners and animals during the use of antimicrobialsusing hand hygiene practices after direct contact with petsavoiding intimate contacts with pets through face licking and sharing a bedregular cleaning of households; andwearing rubber, latex, or vinyl gloves when cleaning urine and droppings from insects, rodents, and petsDifferent strategies of mitigation of health risks related to AMR should be developed for animals kept as pets and animals such as rodents and insects that enter household settings temporarily. Nevertheless, the general approach should include obligatory control of antimicrobial use and their release into the environment from all sources. In the case of pets, represented predominantly but not exclusively by cats and dogs, the use of antimicrobials should rely on guidelines issued to veterinary specialists and recommendations issued to pet owners via veterinary clinics. Given the fact that there are limited antimicrobials available for use in companion animals, the mitigation strategy should be of benefit to both animals and humans. Specifically, such a strategy should include:blocking all potential entry routes , particularly in the colder season when the risk of entering is the highestsealing garbage bins and containerssealing food, including pet foods, to avoid cross-contaminationcleaning the household to remove uneaten parts of foodusing traps and baits in case of a high risk of infestation; andpreventing household cats from going outside to limit the predation, potential rodent-cat transmission, and further cat-human transmission.In cases where rodents are not kept intentionally in household settings (predominantly mice and rats), the following mitigation approach is proposed:collection of dog’s and cat’s droppings and their appropriate disposal to avoid contact with insects  that feed or develop in excrements, acquisition of AMR and its further spread to humans .regular cleaning of householdsThe use of preventive measures to limit insects from entering the household setting ; andelimination of insects in household setting The mitigation strategy for the health risks related to AMR transmission via insects entering the household should include:Given that rodents, insects, and pets might co-exist, a combination of these measures might be required to mitigate the health risks associated with AMR.Further research is needed to address several knowledge gaps in the context of the source-pathway-receptor-impact framework.Several insects, rodents, pets, and human ectoparasites such as lice and fleas remain understudied . Hence, The relative contribution of the intrinsic and induced AMR to the AMR burden in insects, rodents, and pets is unclear. This is because partitioning AMR between the two sources is difficult due to the complex behavior of AMR and methodological limitations. Hence, research is required to develop and validate tools to partition AMR between the two sources.The transfer of AMR in insects, rodents, and pets to humans might occur via multiple pathways including contaminated food ,262, butCell membranes and proteins of some organisms might act as receptors or primers that suppress or amplify AMR ,264. HowEvidence shows that insects are a potential source of insect defensins or antimicrobial peptides that provide the first line of defense against antimicrobials ,267. TheDespite harboring AMR, edible insects, rodents, birds, and other wild animals are sources of food and nutrition in several cultures in South America, Asia, African, and even some developed countries ,135,271 The evidence linking AMR in insects, rodents, and pets remain largely inferential and qualitative. Further studies are required based on QMRA to determine the link between AMR and human health risks directly. Unlike qualitative or inferential evidence, QMRA uses quantitative dose-response models or relationships to estimate the probability of the occurrence of an endpoint . In the Currently, no estimates exist on the contribution of AMR in insects, rodents, and pets to the global AMR burden and the associated health risks. As Dunachie et al.  pointed Public knowledge, perceptions, attitudes, and practices as well as those of human and animal health practitioners  is a key driver of AMR through overuse, misuse, and abuse of antimicrobials. However, little is known about the contribution of the human factor to the transmission of AMR in the environment–animal–human interface.Developing regions, including Africa, are poorly represented in the literature on AMR, particularly with respect to insects, rodents, and pets. Therefore, further research is needed to determine the nature of AMR in various species of insect, rodent, and companion pets in developing countries. Moreover, such research should address several generic knowledge gaps highlighted here.The bulk of studies are based on conventional culturing and molecular techniques. Several recent advances in analytical tools such as computational or in-silico techniques, genomics, network analysis, and big data analytics only received a cursory application in research on AMR in insects, rodents, and pets. Yet, these emerging tools have the potential to complement existing conventional methods and provide further insights into the complex behavior of AMR. The generic applications of emerging and novel analytical tools in AMR research are discussed in an earlier paper focusing on AMR in the funeral industry . Here, pGenomics is a collective term referring to a range of advanced molecular techniques, including, among others, meta-transcriptomics, (exo)metabolomics, metagenomics, and proteomics . CompareIn-silico or computational techniques are collection modeling or simulation tools that allow the rapid computer-aided design of virtual experiments. In the context of AMR in the environmental–animal–human continuum, in-silico techniques have the following potential applications—(1) rapid prototyping and analysis of complex systems including metabolic and AMR networks, which are difficult to study via conventional research methods, and (2) outputs of in-silico analysis can be used to inform the cost-effective design of laboratory and field experiments, and subsequent analysis of data.Does AMR in humans, insects, rodents, and pets originate from the same environmental pool or resistome?’. Network analysis is a powerful tool used to understand complex interactions among various components. Thus, a combination of network analysis and genomics can be used to unravel such complex interactions and reveal the degrees of similarities of AMR in the various compartments or resistomes.The occurrence, dissemination, and fate of AMR in the various compartments form a complex network that is often difficult to disentangle using conventional tools. One question that arises is, ‘A comprehensive understanding of the occurrence and behavior of AMR requires analysis of various media. These include (i) environmental media , (ii) animals including insects, rodents, pets, and their prey and predators, and (iii) human media such as feces, gut specimen, and serum. Such efforts will generate large quantities of data  that are often difficult to analyze and visualize using conventional statistical tools. Big data analytics such as machine learning might provide ideal tools for the extraction of subtle trends and patterns in such large datasets encompassing AMR in environmental, animal, and human media and their potential human and ecological health risks.The application of these emerging tools could significantly improve our understanding of the complex behavior and health risks of AMR. Research applying a combination of these emerging tools represents the next frontier in AMR research in the environment–animal–human continuum.The present review applied the source-pathway-receptor-impact framework to track the occurrence and circulation of AMR in household and edible insects, rodents, and pets. Insects and rodents acquire antimicrobial resistance via foraging of prey harboring AMR, which in turn proliferate and persist in various compartments of insects, rodents, and pets, including the skin and gut systems. The widespread use of antimicrobial treatments in veterinary medicines and pet foods also induce AMR micro-organisms found in pets. Thus, several household insects  and edible ones , rodents , and pets , harbor and act as reservoirs of antimicrobial resistance. To date, antimicrobial-resistant microorganisms and their corresponding ARGs for a diverse range of first-line and last-resort antimicrobials, including antibiotics, are reported in insects, rodents, and pets. Subsequently, household insects, rodents, and pests act as vectors that disseminate antimicrobial resistance into other environmental compartments, including humans via direct contact, as prey for other organisms, human food contamination, and horizontal gene transfer. Thus, household insects, rodents, and pets might act as sentinels or bioindicators for the surveillance of AMR. In this regard, insects, rodents, and pets could be subsequently used as an early warning system for the surveillance of AMR ahead of its detection in humans.Human exposure and health risks, and key risk factors were discussed, including those unique to low-income countries. Current evidence on human exposure and health risks is largely inferential and qualitative. Hence, comprehensive data based on quantitative microbial risk assessment are still lacking, making it difficult to trace specific human health risks or outcomes to AMR in insects, rodents, and pets. To safeguard human health, mitigation measures are proposed based on the one health approach. Future research, including ten knowledge gaps, were highlighted, including (1) quantitative microbial risk assessment, (2) investigating AMR in common household insects, rodents, and pets in developing countries, (3) partitioning antimicrobial resistance in insects, rodents, and pest between anthropogenic and natural sources, and (4) use of emerging and novel techniques  to understand the role of household insects, rodents, and pets in the persistence, circulation, and health risks of AMR. These emerging tools can complement conventional ones to address the highlighted knowledge gaps and improve our understanding of the occurrence and circulation of AMR in the environment–animal–human continuum.


Mechanical property is one of the most important properties of nanofiber membranes. Electrospinning is widely used in the preparation of nanofibers due to its advantages such as good stability and easy operation. Compared with some nature silk, the mechanical properties of nanofibers prepared by electrospinning are poor. Based on the principle of vortex spinning and DNA structure, this paper designed an air vortex electrospinning device that can control the structure of macromolecular chains in nanofibers. When a weak air vortex is generated in the electrospinning process, the macromolecule chains will entangle with each other and form a DNA-like structure so as to improve the mechanical property. In addition, when a strong air vortex is generated during the electrospinning process, the nanofibers will adhere to each other, thereby enhancing the mechanical property and enlarging the pore size. Vortex spinning is a pneumatic spinning method and widely used in the textile industry ,2,3. It Polyvinyl alcohol (PVA) 1799 with an average molecular weight of 200,000 g/mol was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. , and used as received, alcoholysis degree of the PVA was 97.5–99.0 mol%. Polyacrylonitrile (PAN) with an average molecular weight of 150,000 g/mol was purchased from Beijing Lark Branch Co., Ltd.  and used as received. N, N-dimethylformamide (DMF) was purchased from Shanghai Chemical Reagent Co. Ltd. , and used as received.Electrospun nanofiber’s morphology was analyzed using an S4800 cold field scanning electron microscope . To determine the diameter distribution of nanofibers, we randomly selected 500 nanofibers from 50 SEM images, and measured them with ImageJ software , finally calculated the average value. Mechanical property was measured by the material testing machine . The pore size distribution was measured for the nanofiber membranes by a capillary flow porometry . FTIR spectra of nanofiber membranes were obtained on a Fourier transform infrared (FTIR) spectroscopy .1.6 g PVA powders were dissolved in 18.4 g deionized water, then the mixtures were magnetically stirred on the heating magnetic stirrer  at the temperature of 80 °C for 3 h to achieve homogeneous solution. 1.6 g PAN were dissolved in 18.4 g DMF, then the mixtures were magnetically stirred on the heating magnetic stirrer  for 4 h to achieve homogeneous solution.The obtained solution was put into a 10 mL syringe, which was mounted in a syringe pump . A high voltage power supply  was used in our experiment. The voltage was 18 kV, the flow ratio was 1 mL/h, the distance between the needle and the collector was 20 cm, the temperature was 24 °C, and the relative humidity was 37%. We repeated the experiment approximately 15–20 times.Based on the working principle of air-jet vortex spinning, in order to improve the mechanical properties of nanofiber membranes, we used vortex in the electrospinning process for the first time and designed an air vortex electrospinning device. In this part, we discussed the influence of air vortex on nanofibers. PVA and PAN nanofibers were prepared using the air vortex electrospinning device. The electrospinng condition were all the same. The voltage was 18 kV, the flow ratio was 1 mL/h, the distance between the needle and the collector was 20 cm, the temperature was 24 °C, and the relative humidity was 37%.The entanglement of macromolecular chains inside nanofibers will greatly affect the mechanical properties of nanofiber membranes. −1 attribute to C-C vibration, 1100 cm−1 relate to C-O stretching vibration, C-OH vibration at 1410 cm−1, 1710 cm−1 attribute to C=O stretching vibration, CH2 bending and stretching vibration at 2922 cm−1 [−1 (C−1 (C=O of ester group), 1446 cm−1 (C-H bending in CH2) and 1245 cm−1 (C-N bending) are clearly observed [922 cm−1 ,26. The observed ,28,29. FThe pore size distributions of PVA and PAN nonofiber membranes were measured by a capillary flow porometry. It can be seen from In the previous section, we used the air vortex electrospinning device and selected two different polymer solutions for spinning and discussed the influence of the air vortex on the macromolecular chains inside the nanofibers. The previous experiments found that the weak air vortex has a greater impact on the macromolecular chains inside the nanofibers but has almost no effect on the nanofibers. In addition, the use of the air vortex improves the mechanical properties of the nanofiber membranes.In this section, we changed the intensity of the air vortex by selecting different airflow speeds and discussed the influence of air vortex strengths on the macromolecular chains inside the nanofibers and the interaction between nanofibers. In this section, 8% PVA solution was used for spinning, and the speed of the pumped airflow was selected as 1 m/s, 2 m/s, 3 m/s, 4 m/s and 4.5 m/s, respectively. The silk conditions and spinning environment were also consistent with those described above.The change of air vortex strength would have a great impact on the morphology of nanofibers. From the SEM images we know that when the air vortex strength is weak, it just has effect on macromolecule chains, but once it increases to a critical value, the air vortex begins have effect between nanofibers. So, the air vortex strength will greatly affect nanofiber membranes’ mechanical property. It can be seen from The pore size distributions of PVA membranes obtained in different air vortex strength were measured by a capillary flow porometry. From In this paper, for the first time, we use air vortex in the electrospinning process. Compared with traditional electrospinning and cross-certified through the scanning electron microscopy, mechanical property test, Fourier transform infrared spectroscopy and pore size test, we found that weak air vortex in the electrospinning process could control the macromolecule chains’ structure in a nanofiber, making the macromolecule chains entangled with each other and forming a DNA-like structure. As the intensity of the vortex increases, the diameter of the nanofibers gradually decreases. When the vortex strength increases to a certain value, the entanglement of the macromolecular chains inside the nanofiber reaches the limit, and at the same time, the nanofibers begin adhering to each other. As the vortex intensity increases, the mechanical properties of the nanofiber membrane will also increase. Besides, it is also observed that strong air vortex could not only improve the mechanical property of the nanofiber membrane, but also enlarge the pore size, which provide a new and simple method for fabricating high strength and large pore size nanofiber membrane.In a word, this paper proposes a promising method to control the macromolecule chains structure in a nanofiber and provides an effective and simple method to prepare nanofiber membranes with high strength and large pore size.