In gas-phase experiments using molecular beams, formationof many different isomers cannot be prevented, and their presencesignificantly complicates the assignment of spectral lines. Currentisomer-resolved spectroscopic techniques heavily rely on theoreticalcalculations or make use of elaborate double-resonance schemes. Weshow here that isomer-resolved spectroscopy can also be performedusing a single tunable laser. In particular, we demonstrate single-colorisomer-resolved spectroscopy by utilizing electrostatic deflectionto spatially separate the isomers. We show that for 3-aminophenolwe can spatially separate the  I2 or different ordering of macromolecular structures such as proteins.3Chemicaland biological functionality is defined through the underlyingmolecular structure. A special part of this structure–functionrelationship is the role of conformational isomerism, that is, systemsthat can interconvert through the rotation around a single bond. Thisseemingly trivial structural change can have far-reaching consequencesand can lead to, for example, different chemical reactivities6 This wide conformational sampling can be considered a blessing anda curse. The presence of many structures allows for detailed studiesof intramolecular interactions and also aids in linking gas-phasespectroscopy to condensed-phase systems, where typically one conformationdominates due to intermolecular interactions with the solvent. However,extracting spectroscopic information for one particular conformationpresent in the supersonic expansion and assigning this to a specificmolecular structure is not trivial and typically relies on ab initio theory.The occurrence of different conformershas received considerableattention with the advent of high-resolution gas-phase spectroscopybased on supersonic molecular beams. Under the collision-free androtationally cold conditions in these sources, different conformationsdo not interconvert and can be considered separate molecular species.Even for small organic systems, many conformers are frequently observedin the gas phase, for example, the smallest amino acid glycine hasbeen produced in at least four different gas-phase conformations,with the exact number still subject to intense debate.7 These rely on having one resonant laser to selectivelyionize the ground state of a specific conformer and a second laserto drive a transition that depletes (“burns”) the groundstate. The burn laser is fired first, and scanning this thus allowsrecording a conformer-resolved spectrum, which in favorable casescan be assigned to a specific molecular geometry with the aid of theoreticalchemistry calculations. This approach, and variations thereof, isnow widely used across both the IR and UV spectral regions.12The current experimental approachto conformer-resolved spectroscopyis based on double-resonance hole-burning techniques.14 and then record single-colorresonance enhanced multiphoton ionization (REMPI) spectra of the conformer-puresample. This allows the trivial and unambiguous assignment of theobserved spectral lines to a molecular structure. We demonstrate thishere for the syn and anti-conformersof 3-aminophenol, which have a difference in dipole moment of around1.5 D, as shown in We present here an alternative approach to record conformer-resolvedelectronic spectra that does not rely on double-resonance techniquesand does not require the calculation of theoretical spectra. In particular, we show that we can make use of the differencesin the permanent dipole moment of conformers to fully separate themusing electrostatic fieldsII15 operated at 500 Hz and heated to 140 °C.The molecular beam was skimmed twice before entering a 300-mm-longelectrostatic deflector. This had a rod-and-through-type electrodegeometry in which the electrodes were separated by a distance of 3.4mm.16 The electrodes were designed suchthat the field gradient is nearly constant in the vertical directionwhile being almost zero in the horizontal direction. Polar moleculesflying through the deflector therefore experience a force in the verticaldirection, depending on their effective dipole moment.14 The deflector was operated at potential differencesup to 15 kV, corresponding to a maximum field gradient of around 50kV/cm. Following the deflector, the molecular beam was skimmed oncemore before entering the interaction chamber. Further technical detailsare given in the Supporting Information. In the detection region, molecules were ionized via 1 + 1 resonanceenhanced multiphoton ionization (REMPI), using the frequency doubledoutput of a Nd:YAG pumped dye laser (Pyrromethene-597 dye pumped at532 nm), providing 7 ns pulses that were attenuated to 6 μJper pulse, and operating at 50 Hz. The laser was focused using an f = 750 mm lens to a spot size of approximately 100 μminside the center of a three-plate velocity-map imaging spectrometer,17 operated here in ion time-of-flight mode. Spatialdistributions of the molecular beam were measured by translating thelaser focus through the molecular beam.3-Aminophenol  was used without furtherpurification and introduced into a molecular beam through seedingin 28 bar of neon and expansion in a pulsed Even Lavie valve,18 Trajectories of individual quantum states  were then propagatedthrough the experimental setup. Individual trajectories were combinedto a simulated deflection profile through Boltzmann weighting, withthe rotational temperature fitted to the experimental data.14Experimental deflectionmeasurements were complemented by trajectorysimulations. For these, the Stark effect of both conformers was calculatedusing the freely available CMIstark software package.III–1 is shownat the top of 19 with band origins forthe syn and anti conformers at 34 106.8and 34 473.3 cm–1, respectively. Assumingidentical efficiencies of the REMPI process for both conformers, theorigin peaks yield a syn/anti ratio of 1:2.6. A plethora of peaksfrom vibrational transitions is furthermore observed across this wavenumberregion. In order to unambiguously assign these, we turn to the electrostaticdeflection technique.A conventional single-color 1 + 1 REMPI spectrum of 3-aminophenolfrom 34 100 to 35 000 cmsyn and anti conformers was measured at potential differences of 0 and 15 kVby measuring the intensity of the respective origin bands while scanningthe laser height. In syn conformation,while ∼0.8 mm below the center the molecular beam was almostfully depleted for the syn conformer. The inset of syn conformer across the molecularbeam deflection coordinate for a potential difference of 15 kV acrossthe deflector. The dashed lines indicate the positions where the mostpure samples of the syn and anti conformers can be probed, while still having reasonable intensity.Applying a potential differenceacross the deflector electrodesleads to a shift and dispersion of the species within the molecularbeam, depending on the underlying effective dipole moment to massratio. The deflection of both syn and anti conformers are shown in syn spectrum (panel b) subtracted. It is clear from thesespectra that the deflection technique creates conformer-pure samples,making assignment of lines to individual conformers trivial. Thisis further highlighted in –1. The collectedspectrum for the anti conformer contains a smallcontribution from the syn conformer, but the muchreduced intensity still allows unambiguous assignment. Moreover, thevery pure spectrum obtained for the syn conformerallows reconstruction of the pure-anti spectrum bysubtracting the syn contribution from the collectedmixture spectrum, as shown by the green line in REMPI spectra for these pure syn and anti conformers, along with tentative assignments,are given in the Supporting Information. Assignments are based on previously published IR–UV doubleresonance spectroscopy experiments performed at the FELIX free-electronlaser facility and the corresponding high-level theory,20 as well as a previously published UV REMPI spectrum.21 We observe several features that have not been experimentally observedor assigned previously. For example, we observe clear spectral featuresdue to an aromatic ring out-of-plane bend at 217 and 227 cm–1 for the syn and anti conformers,respectively. This was not previously observed but agrees well withpublished theory.20 We furthermore observedstrong bands at 322 cm–1 (syn)and 314 cm–1(anti), which theorysuggests are due to OH wagging or NH2 torsion. The previousexperiments at FELIX suggest that the OH wagging shows a significantlystronger response, and we tentatively assign the observed bands tothis. Interestingly, in the S1 state probed here, the bandbelonging to the syn conformer appears higher inenergy, whereas in the S0 state accessed by double-resonanceexperiments, the anti band is higher two tunable laser sources. Moreover, schemes based on hole burningfrequently require IR “burn” pulses with significantpulse energy to achieve sufficient depletion, which are not widelyavailable and restricted to a few IR free-electron laser sources aroundthe world. The electrostatic separation approach furthermore allowsassignments of spectral lines to individual conformers without relyingon calculations, i.e., without any 22 and also within reach of new approachesto buffer-gas cooling that could provide significantly slower molecularbeams, and hence increased electrostatic deflection.23 We also note that, in principle, the production of isomer-puresamples is not required, as long as samples with different (but known)population ratios can be produced, the individual spectra can be recoveredthrough a global analysis.24Our methodology is widely applicableand requires only that moleculescan be entrained into a rotationally cold molecular beam, and thatthe isomers have a sufficient difference in dipole moment. For typicalaromatic molecules with masses up to a few hundred Daltons, a differencein dipole moment of ∼1 D and a rotational temperature of afew Kelvin is sufficient to obtain a pure sample of the more polarisomer. The latter is routinely achieved in pulsed supersonic molecularbeams,25 For example, it would allow the separation and individual studyof different tautomeric structures such as those of cytosine, whichis known to exist in six tautomeric forms, of which three typicallyare present in a molecular beam.26 Thesethree tautomers have dipole moments of 6.15, 4.52, and 3.10 D, whichis sufficient to separate and study these structures individually.In general, spatial separation of isomers in a molecular beam addsisomer specificity to techniques that by themselves lack the spectralresolution needed to distinguish isomers. This has been demonstratedfor molecular collision experiments1 andis furthermore of use to any ultrafast dynamics experiment, which,due to the inherent bandwidth in a femtosecond pulse, cannot distinguishisomers spectroscopically.Whilewe demonstrated here the collection of pure spectra for individualconformers , the approach is generally applicableto all species within a cold molecular beam, from other forms of structuralisomerism to solvent–solute clusters formed during expansion.IVsyn and anti conformers of 3-aminophenol,and fully isomer-resolved REMPI spectra were presented and assigned,including several previously unobserved lines. The approach is widelyapplicable to any polar molecule with a sufficient difference in dipolemoment between the isomers. It is furthermore not limited to single-colorREMPI spectroscopy as adopted here, instead the physical separationof the isomers in principle allows probing by any laser-based spectroscopicmethod. When using REMPI, it requires only a single resonance process,in contrast to hole-burning approaches that are double-resonance spectroscopies.Its implementation requires only a static electric field and allowsone to add isomer specificity to any molecular-beam-based spectroscopyexperiment.We presentedhere a technique for performing isomer-resolved spectroscopyusing only a single table-top laser, and which allows the assignmentof transitions to an isomer without relying on theoretical predictionsor double resonance schemes. This was demonstrated for the


However, since different research efforts within machine ethics have discipline-specific concepts, practices, and goals, the resulting body of work is pestered with conflict and confusion as opposed to fruitful synergies. The aim of this paper is to explore ways to alleviate these issues, both on a practical and theoretical level of analysis. First, we describe two approaches to machine ethics: the philosophical approach and the engineering approach and show how tensions between the two arise due to discipline specific practices and aims. Using the concept of disciplinary capture, we then discuss potential promises and pitfalls to cross-disciplinary collaboration. Drawing on recent work in philosophy of science, we finally describe how metacognitive scaffolds can be used to avoid epistemological obstacles and foster innovative collaboration in AI ethics in general and machine ethics in particular.Recent advancements in artificial intelligence (AI) have fueled widespread academic discourse on the ethics of AI within and across a diverse set of disciplines. One notable subfield of AI ethics is  Grand challenges are complex problems of global concern that call for inter- and transdisciplinary research efforts  is a subfield of AI ethics that seeks to endow artificial systems, software and hardware alike, with ethical faculties —the conceptual exploration of what computer systems ought to do, and correspondingly, what systems ought to be built.The engineering approach to machine ethics (EME)—the exploration of what kind of morality can be implemented in computer systems, and what moral systems can be built.The majority of work in ME is approached from two disciplinary families, namely moral philosophy and computer science , can have a moral agency or status , the EME drowns in an ocean of semantic confusion. While philosophy can allow for a certain interpretative headroom  and disagreement , “empathic rationality” capable of moral imagination and reflective equilibrium  ethical machine, one would necessarily start from the assumption that it is in fact possible to do so. As a result, one might commit to computationalism about cognition and properties necessary for morality; not because it is the most compelling theory, but because it works in favor of one’s epistemic aim. Furthermore, what seems like trivial premises for some disciplines, might be conceived as disrespectful or even harmful for others; e.g., ignoring the results of millennia-old debates, failing to engage at a normative or societal level, or disregarding what is technically feasible, scientifically explainable, or empirically supported.do good  or be good . There is also an important difference between ethical theory as normative ideal and in terms of action-guidance , it is often influenced by broader and less salient outlooks and assumptions stemming from one’s disciplinary perspective, e.g., about what AI is and what it should be.why AMAs are desirable or useful and for whom. For instance, Van Wynsberghe and Robbins  identifying and avoiding disciplinary capture; and (ii) articulating the underlying views that supports conflicting perspectives on machine morality .In summary, this paper supports three claims:


Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma. It is a clinically and morphologically heterogeneous entity that has continued to resist complete subtyping. Molecular subtyping efforts emerged in earnest with the advent of gene expression profiling (GEP). This molecular subtyping approach has continued to evolve simultaneously with others including immunohistochemistry and more modern genomic approaches. Recently, the veritable explosion of genomic data availability and evolving computational methodologies have provided additional avenues, by which further understanding and subclassification of DBLCLs is possible. The goal of this review is to provide a historical overview of the major classification timepoints in the molecular subtyping of DLBCL, from gene expression profiling to present day understanding. Diffuse large B-cell lymphoma (DLBCL) is the most common non-Hodgkin lymphoma in the western hemisphere . DiagnosIt was not long ago that the diagnosis of liquid tumors rested on morphology, immunohistochemical analyses, and cytogenetics. The original diffuse large B-cell lymphoma (DLBCL) molecular subclassification followed the advent of DNA microarrays, a technology that allows the analysis of thousands of expressed genes simultaneously; see Although the study population was small by current standards, with imperfect subtyping, the classification was adopted and continues today as the standard of diagnostic care, recognized within the World Health Organization’s Revised Fourth Edition of the Classification of Tumors of the Hematopoietic and Lymphoid Tissues  as germiDespite the reasonable success of standard CHOP and R-CHOP, and the growing number of precision therapies, heterogeneity remained an issue, as evidenced by inconsistent treatment responses and relapses despite COO subtyping. Although gaining favor and resolving power, it was thought perhaps that the transcriptionally based GEP was an inadequate representation of underlying aberrant genetic programming . However, practically, other technical hurdles, such as the logistical necessity for fresh tissue specimens, stood in the way of immediate clinical adoption of GEP in the subcategorization of DLBCLs. Initially, this was a difficult step to overcome, however several novel testing options were eventually able to surmount this issue, including Nanostring , HTG 1010, and RMYC aberrations along with MYC IHC over-expression. Other studies using mouse modelling also supported the molecular ideas emerging from GEP, showing oncogenic driver mutations in genes such as EZH2 and MYD88 that promoted lymphoma development . It. It66]. MYC and BCL2 and/or BCL6, helps to identify double-hit or triple-hit, high grade B-cell lymphoma with even poorer outcome and the need for more intensive treatment plans [In countries with limited resources, following H & E review, an immunohistochemistry panel with the markers for CD20, CD5, CD21, and Ki67 reaches a correct diagnosis of DLBCL in 83% of the cases . Cytogennt plans . After an initial histopathological workup, the specimen could be analyzed using a targeted sequencing panel of a limited set of genes either by sending to a sequencing company or to a central hematopathology laboratory. A more centralized or standardized approach would allow collection of additional data regarding predictive biomarkers, and eventually attribution to clinical trials. The classification of DLBCL has come a long way, and molecular techniques have provided a wealth of information to the underlying pathophysiology, prognostic, diagnostic, and therapeutic implications. Even at the time of writing and publishing this review, there will be undoubtedly be updates, if that is any indication of the speed of development in molecular techniques and novel technology that are able to further enhance our understanding of this heterogenous and diverse lymphoma.


Alcohol use remains a major public health concern and is especially prevalent during adolescence. Adolescent alcohol use has been linked to several behavioral abnormalities in later life, including increased risk taking and impulsivity. Accordingly, when modeled in animals, male rats that had moderate alcohol consumption during adolescence exhibit multiple effects in adulthood, including increased risk taking, altered incentive learning, and greater release of dopamine in the mesolimbic pathway. It has been proposed that alcohol arrests neural development, “locking in” adolescent physiological, and consequent behavioral, phenotypes. Here we examined the feasibility that the elevated dopamine levels following adolescent alcohol exposure are a “locked in” phenotype by testing mesolimbic dopamine release across adolescent development. We found that in male rats, dopamine release peaks in late adolescence, returning to lower levels in adulthood, consistent with the notion that high dopamine levels in adolescence-alcohol-exposed adults were due to arrested development. Surprisingly, dopamine release in females was stable across the tested developmental window. This result raised a quandary that arrested dopamine levels would not differ from normal development in females and, therefore, may not contribute to pathological behavior. However, the aforementioned findings related to risk-based decision-making have only been performed in male subjects. When we tested females that had undergone adolescent alcohol use, we found that neither risk attitude during probabilistic decision-making nor mesolimbic dopamine release was altered. These findings suggest that different developmental profiles of the mesolimbic dopamine system across sexes result in dimorphic susceptibility to alcohol-induced cognitive and motivational anomalies exposure. Alcohol remains the most frequently used substance among adolescents, and in this age group, there is a prevalence of high levels of binge drinking . Therefore, we tested evoked mesolimbic dopamine release across developmental time points, using stimulation procedures that discern neuronal terminal or cell-body mechanisms of potential age differences.ad libitum except as noted. All work in this manuscript was approved by the Institutional Animal Care and Use Committee of the University of Washington.Sprague-Dawley rats  began experimental procedures at Post Natal Day (PND) 25 (gavage) or 27 (gelatin) for animals exposed to alcohol, or PND 30, 50, or 120 for neurochemical experiments without alcohol exposure. Rats were housed in polycarbonate tubs on a 12-h light/dark cycle (lights on a 06:00) for one week before this date. Water and rodent chow  was available ad libitum access to food and water. Gel intake levels were measured daily and expressed in g/kg of body weight. All rats had access to only control gelatin for the first three days; after which rats were divided into either ethanol or control gelatin groups matched by weight and baseline intake for 20 days of assigned gelatin intake.Alcohol administration through the voluntary consumption of gelatin containing alcohol was presented to adolescent rats (PND 30–50) in a gel matrix consisting of distilled water (76.67%), Knox Gelatin (3.33%), polyose (10%), and 190 proof ethanol (10%), whereas the control gelatin contained distilled water in place of ethanol. Preparation was as described . It is important to mention that no overt signs of withdrawal were observed. Once the withdrawal period was completed, the rats began a food restriction diet of 90 ± 2% of their bodyweight and were exposed to 45 mg sucrose pellets  in their home cage to reduce neophobia. Additionally, prior to the start of the behavioral tasks, the rats underwent one magazine-training session in a standard operant chamber  where they were given 15 min to consume 10 sucrose pellets in the magazine tray.Risk attitude was tested in female and male rats using a probabilistic decision-making task. After magazine training was completed, rats were trained on an operant fixed ratio (FR) schedule to a criterion of >23 presses out of 20 total trials where one pellet was delivered following the depression of the left or right lever. Rats then underwent auto-shaping, where the rats were required to first nose-poke the magazine tray to begin the trail where the intertrial interval was increased from 0 to 15 s, the time to perform the trail initiating the poke was decreased to 10 s, and the intertrial interval was increased to 30 s.Detailed methods for these and the following tasks can be found in previous publications  and medial forebrain bundle (MFB) stimulated dopamine release in the nucleus accumbens (NAcc) as follows.To test dopamine transmission across adolescent development, dopamine release was evoked by MFB or PPT stimulation and measured with FSCV during non-survival surgeries in females and males during early adolescence (PND 30–35), late adolescence (PND 50–55) or adulthood (PND 120–125). Female adult rats from control and gelatin decision-making behavioral groups were also tested this way.Rats were anesthetized with a 1.5 g/kg urethane (i.p.) and head-fixed in a Kopf stereotaxic instrument. The skull was exposed and burr holes were drilled targeting the NAcc , and PPT . Another burr hole was drilled on the contralateral side for placement of the reference electrode (Ag/AgCl). A carbon-fiber microelectrode was centered above the NAcc burr hole and lowered 6.8–7.2 mm ventral from the top of the brain.p < 0.05) between working electrode placement in the NAcc  group = 0.522; medial/lateral: F group = 0.9984; anterior/posterior: F group = 1.275), stimulating electrode placement MFB  group = 0.7424; medial/lateral: F group = 0.7336; anterior/posterior: F group = 0.8156) or stimulating electrode PPT  group = 0.7081; medial/lateral: F group = 1.315; anterior/posterior: F group = 0.4166) between experimental groups.On completion of the experiment, current was passed through the voltammetry electrode to produce a lesion to aid histological identification of the recording location. Animals were then sacrificed using phenytoin/pentobarbital (Bethanasia) and their brains were harvested for histological analysis of the recording and stimulating electrode placement. On analysis of these data using one-way analysis of variance (ANOVA), there were no significant differences in electrode placement between experimental groups. Specifically, no differences were observed  at a rate of 400 V/s and a 10 Hz .p < 0.05 within correction for multiple comparisons as appropriate.Stimulated dopamine release was analyzed with two-way mixed-measures ANOVA with stimulation current or the number of pulses as a within-subject, and age or treatment as a between-subject factor. Behavioral data for decision-making sessions were analyzed using a two-way mixed-measures analysis of variance using probability as a within-subject and treatment as a between-subject factor. For intragastric alcohol administration, data were analyzed using three-way mixed-measures ANOVA with probability as a within-subject and treatment and sex as between-subject factors. All data are presented as mean ± SEM and threshold for statistical significance was set at F current = 93.25, p < 0.0001, F current = 80.27, p < 0.0001, F current = 31.79, p < 0.0001, F current = 35.09, p < 0.0001, F age = 0.2164, p > 0.05, F current × age = 0.5093, p > 0.05, F age = 0.5778, F current × age = 0.3545, p > 0.05, F age = 0.7016, F current × age = 0.4490, p > 0.05, F age = 0.4182, F current × age = 0.3201, p > 0.05, To investigate developmental changes in the excitability of mesolimbic dopamine neurons, we measured dopamine release in the NAcc in rats aged 30 (early adolescence), 50 (late adolescence), or 120 (adulthood) days postnatally. First, we evoked dopamine release with electrical stimulation of the MFB using a range of stimulation parameters. Dopamine release was reliably detected, with increasing peak dopamine concentration to higher simulation current or number of pulses in males  current = 35.27, p < 0.0001, F current = 29.33, p < 0.0001, F current = 30.13, p < 0.0001, F current = 56.40, p < 0.0001, F age = 3.157, p > 0.05, F age = 2.413, p > 0.05, F current × age = 2.598, p < 0.01, F current × age = 2.572, p < 0.01, F age = 1.249, p > 0.05, F current × age = 0.7699, p > 0.05, F age = 1.823, p > 0.05, F current × age = 1.267, p > 0.05, F sex = 2.431, p > 0.05, F age = 0.4258, p > 0.05, F sex × age = 0.1330, p > 0.05, F sex = 0.9110, p > 0.05) or age  age = 1.552, p > 0.05), but there was a significant interaction between these factors  sex × age = 4.444, p < 0.05, Stimulation of the MFB activates ascending dopamine axons and provides an assessment of presynaptic  function in the control of dopamine release. To also assess the excitability of dopamine neurons to afferent input, we stimulated the PPT to evoke dopamine release by transsynaptic stimulation. PPT stimulation consistently increased dopamine release in a current and pulse-number sensitive manner in males  current = 51.09, p < 0.0001, F pulses = 70.77, p < 0.0001, F current = 22.71, p < 0.0001, F pulses = 36.96, p < 0.0001, F treatment = 1.031, p > 0.05, F current × treatment = 0.5739, p > 0.05, F treatment = 0.05355, p > 0.05, F pulse × treatment = 0.3920, p > 0.05, F treatment = 0.03726, p > 0.05, F current × treatment = 1.203, p > 0.05, F treatment = 0.2302, p > 0.05, F pulse × treatment = 0.2803, p > 0.05, Previously, we observed increased dopamine release during adulthood in male rats that consumed alcohol during adolescence  probability = 31.40, p < 0.0001, F treatment = 0.2968, p > 0.05, F treatment × probability = 0.7999, p > 0.05, This failure to observe enduring changes in dopamine release following AAU in females provides an additional challenge on previous interpretations of AAU on cognition. The elevated dopamine in males is concomitant with an increased risk attitude during probabilistic decision making. However, the lack of elevated dopamine release in females questions whether the same behavioral changes would take place. Consequently, we trained female rats with AAU history (gelatin exposure described above) to perform probabilistic decision-making where animals choose between deterministic small rewards (two food pellets) and probabilistic large rewards (four food pellets). The probability of delivery of the large reward when chosen was systematically descended during each behavioral session . The frequency of choosing the large reward varied according to its probability of delivery (n = 12) of alcohol during adolescence. This value is considerably higher than male rats who underwent comparable training. For example, Schindler et al. (t(28) = 9.833, p < 0.0001, unpaired t-test vs. females in the current study). Nonetheless, this model results in only moderate blood ethanol concentrations  probability = 20.91, p < 0.0001, F probability = 22.35, p < 0.0001, F treatment = 0.5446, p > 0.05, F probability × treatment = 0.1471, p > 0.05, F treatment = 7.764, p < 0.01, F probability × treatment = 0.6704, p > 0.05, F sex = 5.920, p < 0.05). These data demonstrate that, regardless of the method of ethanol administration and the amount received, AAU leads to risky decisions making in adult males, but not in females.Females that underwent the gelatin AAU model consumed 30.32 ± 1.96 g/kg/day (r et al.  reportedHere, we measured dopamine excitability across normal adolescent development. The experiments were designed to provide a platform to investigate the hypothesis that behavioral perturbations in adulthood arising from AAU are caused by alcohol-arresting post-adolescent neural development.We observed a significant peak in dopamine neuronal excitability during the adolescent period in males. Therefore, if AAU arrests the development of this system then we would anticipate higher levels of dopamine during adulthood than in controls where dopamine excitability drops following adolescence. This prediction is consistent with our previous work demonstrating greater dopamine excitability during adulthood in animals that underwent AAU as compared to controls (Schindler et al., Conversely, in females, we did not observe changes in dopamine excitability across development. This lack of a peak in adolescent females was surprising, and seemingly refutes the idea that altered behavior in later life could be a result of arrested development of the mesolimbic dopamine system. However, this position assumes that AAU increases risk attitude during probabilistic decision-making in females like it does in males (Nasrallah et al., Our findings identify clear sex differences in the mesolimbic dopamine system during adolescent development and their correlation with the effect of AAU on later life physiology and behavior. However, it is not clear whether the dimorphic effects of alcohol are categorical, or whether they are quantitative effects. With regard to the dose of alcohol, females consumed a larger quantity of alcohol, during gelatin-based self-administration, than males in previous studies (Nasrallah et al., Sex differences in the enduring effects of AAU have previously been observed (Bava and Tapert, The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The animal study was reviewed and approved by University of Washington Institutional Care and Use Committee.LK, AS, and JC conceived the studies. AA, LK, and AS performed the studies. AA analyzed the data. AA and PP wrote the manuscript. AA, LK, AS, and PP edited the manuscript. All authors contributed to the article and approved the submitted version.


In this study, a modified version of electrospun nylon 66 nanofibers by silica particles were blended into ordinary Portland cement to investigate the microstructure and some mechanical properties of cementitious material. The addition of silica into the nanofibers improved the tensile and compressive properties of the hardened cement pastes. The observations from the mechanical strength tests showed an increase of 41%, 33% and 65% in tensile strength, compressive strength, and toughness, respectively, when modifying the cement pastes with the proposed nanofibers. The observations from scanning electron microscopy and transmission electron microscopy showed the morphology and microstructure of the fibers as well as their behaviors inside the cement matrix. Additionally, X-ray diffraction and thermal gravimetric analysis clarified the occurrence of the extra pozzolanic reaction, as well as the calcium hydroxide consumption by the attached silica inside the cement matrix. Finally, the observations from this study showed the successful fabrication of the modified nanofibers and the feasibility of improving the tensile and compressive behaviors of cement pastes using the proposed electrospun nanofibers. Today, cement is a basic material that plays an important role in the construction sector. However, there is an increased demand for higher cement strength, due to the feasibility of reducing structural size and cement consumption, which can lead to significant ecological improvement and financial efficiency. Therefore, it has posed a considerable challenge for researchers and manufacturers. Cement matrix strength can be affected by different factors, such as the component ratio, the cement hydration process, the change in the cement matrix microstructure, the curing condition, the addition of additives, and so on ,2. TradiFurthermore, recent findings showed significant performance of different nanosized fibers in improving the cement matrix. The findings by Liu et al.  showed tType I ordinary Portland cement was purchased from Ssangyong Co., Korea . The proThe polymer solutions were prepared in two steps: (1) The solvent was prepared by stirring formic acid and chloroform in a volume ratio of 4:1. (2) Precursors, silica particles, and nylon 66 were added to the prepared solvent to obtain the polymer solutions at a precursor-to-solvent ratio of 1:4 by weight of the polymer solution. This proportion was determined after conducting a trial-and-error process to determine the optimal and spinnable viscosity of the polymer solution using a size 20 Taylor cone. To be more specific, 13 wt% of silica was added to two bottles containing 80 wt% of solvent. Then, the solutions were ultrasonicated for 2 h under laboratory conditions to break the van der Waals forces between the nanomaterial molecules and obtain homogeneous dissolution in the final solutions ,22. FinaElectrospun nanofiber fabrication and the blending process to obtain the composite cementitious materials containing electrospun nanofibers have been reported in previous works ,16. AfteOwing to the changes in the microstructure of the matrix and the difficulty in controlling the spontaneous cracking that occurs during the curing process, it is difficult to conduct a study on the mechanical properties of hardened cement pastes. Therefore, the cement paste samples were designed with small dimensions according to the specifications in the ASTM C307  and ASTMIn this study, the tensile strength and compressive strength tests were evaluated using a 5 kN mortar tensile strength test device in compliance with ASTM C307 and a 1000 kN hydraulic universal testing machine in compliance with ASTM C109/109M, respectively.The FE–SEM analysis was conducted using a Zeiss Merlin Compact system. Input parameters, a working distance of 9 mm, and an acceleration voltage of 5 kV were used. In addition, to increase the observation quality, a 5 Å platinum cover was coated on the sample surfaces.The FE–TEM analysis was conducted using an FEI Tecnai F30 Twin system under an acceleration voltage of 300 kV.XRD analysis was conducted using the D8 Advance system with a scanning speed of 0.4 s/step, a step size of 0.02° (2θ)/step from 5° to 70°, and CuKα radiation .TGA was conducted using a TA instrument SDT-Q600 under a nitrogen atmosphere with a flow rate of 100 mL/min, a heating velocity of 10 °C/min, and a heating range from 25 °C to 1000 °C.The results of the tensile and compressive strength tests indicate that the electrospun nylon 66 nanofibers modified with silica particles are effective in improving some select mechanical properties of the hardened cement paste see . OverallIn the case of compressive strength, an increase of 33% was observed with the addition of NS-N66 NFs, as compared to that of the unmodified paste a. In the4.2H2O), hillebrandite (Ca2SiO3(OH)2) and some dehydrated clinker grains, belite (C2S), and alite (C3S). The hydration products of all the samples were consistent and suitable with the results obtained from Jiang et al. [The XRD patterns of the plain cement and the electrospun nanofiber-blended cement are shown in g et al.  (Figure g et al. ,32.The decrease–increase tendency of the CH and CSH contents of the three samples was consistent with the order of the observations from the mechanical strength tests. The XRD analysis showed that the CH phase in the matrices was consumed. The increase in the CSH content was observed thanks to the pozzolanic reaction between the attached NS and the generated CH phase from the hydration processes.2 from the ambient environment. As can be observed from the TGA results, the CSH content from the nanofiber-blended cement pastes was higher than that of the plain paste, whereas that of CH was lower. It is also worth noting that despite the decomposition range of the CH phase being similar to the decomposition range of the electrospun nanofibers from 400 °C to 500 °C, as presented above, the CH content of the hardened modified cement pastes still decreased compared to that of the hardened plain paste, as indicated by TGA analysis. Consistent with the XRD results, these observations showed a decreasing tendency of the CH and CSH contents in the presence of NS-N66 NFs in the cement. Therefore, the attached silica particles on the nylon 66 nanofibers influenced the final product of the cement paste. Above all, these observations confirmed the initial hypothesis regarding the role of the pozzolanic agent in the attached silica particles.This study determined the influence of electrospun nylon 66 nanofibers containing silica particles on the microstructure and selected mechanical properties of the hardened cement pastes. The following conclusions were drawn:An increase of 41%, 33%, and 65% in tensile strength, compressive strength, and toughness, respectively, was observed when modifying the cement pastes with the proposed nanofibers containing silica particles.The nanosilica particles attached along the axis of the nylon 66 nanofibers were effective in increasing the generated CSH content from the pozzolanic reaction. In addition, the bridging and filling effects of the nanofibers inside the matrix were observed, which increased the mechanical strength of the hardened cement pastes.The XRD and TGA results demonstrated the decrease–increase tendency of the CH and CSH contents when blending the cement with NS-N66 NFs. Therefore, the role of the attached silica particles is clarified.Finally, electrospun nylon 66 nanofibers containing silica particles is a promising candidate for strengthening cementitious materials. It can be seen that an improvement in the mechanical properties of cement can lead to extending the durability and service life of concrete. However, for the practical application of this product, further research needs to be conducted to improve the performance of the composite cement fabrication process, as well as investigate the effectiveness of these nanofibers on other properties of cement paste and concrete. Additionally, the analysis of the life cycle and the life cycle cost of the concrete structure utilizing the proposed cement is necessary, which can evaluate the competitiveness of the proposed cement solution and help authorities in the decision-making process . In addi


Salmonella Reading in Canada and the USA have implicated raw turkey products. To better understand the epidemiology of AMR in farmed turkey production, a scoping review was conducted. The objectives were to identify (1) modifiable factors potentially associated with antimicrobial-resistant Campylobacter, Enterococcus, Escherichia coli and Salmonella enterica along the farm-to-fork pathway in turkeys, and (2) data gaps with respect to factors potentially associated with AMR and Canadian commercial turkey production. A comprehensive search of the peer-reviewed literature was conducted in 2019 and updated in 2021. Thirteen references were included, reporting 36 factors. Antimicrobial use factors and their potential associations with AMR were most frequently reported , followed by biosecurity  and management practices . This review revealed important data gaps; no factors pertaining to S. enterica or to stages other than the farm  were identified, and only one Canadian reference was identified. These findings will inform priorities for future research and surveillance regarding turkeys and AMR.Antimicrobial resistance (AMR) is a complex problem that is a threat to global public health. Consumption of turkey meat may be an important source of foodborne exposure to resistant bacteria; recent outbreaks of multi-drug-resistant  Salmonella Heidelberg human isolates [Antimicrobial resistance (AMR) is a current global threat to public health . Losses isolates . Transmiisolates . Humans isolates . Addressisolates .Salmonella enterica serovars Reading [S. Reading linked to raw turkey products occurred between 2017 and 2020 in Canada, and between 2017 and 2019 in the USA [Farmed turkeys (referred to as turkeys herein) are an important food animal species in Canada, encompassing commercial turkeys, which are slaughtered for meat, and breeder turkeys, which supply hatching eggs destined for commercial turkey production. Turkey meat is the fourth most commonly consumed meat  in Canada, and ranks second for consumption as deli meat . Further Reading , 11, Sch Reading , Heidelb Reading , 14 and  Reading , some of Reading . Notably the USA , 11.Although it is well-established that AMU creates conditions for the emergence, dissemination and persistence of AMR, other factors may influence AMR in the farm-to-fork pathway, such as housing conditions at the farm, or carcass chilling during meat processing , 16. OveCampylobacter, Escherichia coli and S. enterica on-farm isolates were resistant to at least one antimicrobial class [Campylobacter species and composition of Salmonella serotypes [vs. organic or raised without antibiotics flocks) and animal health parameters  vary substantially between poultry species raised in Canada [Existing knowledge synthesis literature on factors potentially linked with AMR in livestock has focused on cattle, chickens and pigs , 20, 21.al class . This inal class –24. Revial class , 26 or hal class . Synthesal class  or utilial class . CIPARS erotypes . As welln Canada –34. On aCampylobacter species, Enterococcus species, E. coli and S. enterica along the farm-to-fork pathway in turkeys, and (2) to describe data gaps in this literature with a particular focus on Canadian commercial turkey production. A factor was defined as a measured observation with an investigated potential association or relationship with AMR, such as AMU at the farm or the use of disinfectants at the abattoir [Given apparent gaps in the AMR literature regarding turkeys, a scoping review was conducted. The objectives were: (1) to qualitatively describe the available peer-reviewed literature reporting modifiable factors potentially associated with the occurrence of antimicrobial-resistant abattoir .This scoping review was conducted as part of a larger project: the Integrated Assessment Modelling of Antimicrobial Resistance (iAM.AMR) project . The goaThis scoping review adheres to the PRISMA-ScR  guidelines for the reporting of scoping reviews . A protoThe research team had expertise in AMR, food safety, epidemiology, veterinary medicine, poultry, library science and synthesis research methodologies. As the initial search was designed to capture data for all facets of the iAM.AMR project , the search was broader than necessary for this review, yet appropriate to address the specific objectives of this review.The initial search was run on 11 April 2019 in five databases: Medline, Embase, Agricola, Centre for Agriculture and Biosciences International Abstracts (CAB Abstracts) and Food Science and Technology Abstracts . For this review, the search was updated on 11 August 2021, where the host species was limited to turkeys . All Supplementary material is available online on the Cambridge Core website.All citations retrieved were imported into reference management software , where exact matches were de-duplicated automatically. Citations were then imported into the web-based systematic review software Rayyan . All potIn vitro studies, reviews and other synthesis methodologies  were excluded. Analytic studies were eligible, including observational and experimental studies.The eligibility criteria are outlined in References reporting a modifiable factor potentially linked with the occurrence of resistance were included if resistance was reported quantitatively in such a way that an odds ratio could be calculated (to account for the needs of the larger iAM.AMR project)  Figs 1 . PotentiReferences only reporting modifiable factors potentially linked with the occurrence of multiple drug resistance in such a way that resistance to individual antimicrobials or antimicrobial classes could not be parsed out were excluded . HoweverFurthermore, references only reporting factors analysed as continuous variables, rather than categorical variables, were excluded given the objectives for this stage of the overall model development for the larger iAM.AMR project . The eliPrimary (title/abstract) screening was performed in Rayyan, and secondary (full-text) screening was performed in Excel , using the criteria given in At each stage, citations or references were screened independently by two reviewers. Conflicts were resolved by consensus and where consensus could not be reached, citations or references were screened by a third reviewer. If an abstract was not available at primary screening, the citation was included and the full text was sought in preparation for secondary screening. At secondary screening in the initial search, selections of ‘unsure’ were either (1) resolved as conflicts, where references were included if both reviewers were unsure, or (2) resolved by a third reviewer, where references were included if the third reviewer was unsure. References were omitted from secondary screening if: the full text was not available, the reference was published in a language other than English, the reference was provided in an alternate format (e.g. conference abstract) or the reference was not unique .References included at secondary screening were imported from Excel into a custom data extraction tool . Characteristics of the references, factors and potential associations with resistance were entered into the data extraction form provided in this database (Supplementary Table S11). Each reference was extracted by a single extractor.Because the initial search included other species in addition to turkeys, it was necessary to identify all references pertaining to turkeys (hereafter referred to as turkey references). Bibliographic data for all references included at secondary screening from the initial search were imported as an Excel file into R , where a custom script was used to identify potential turkey references based on the presence of at least one of the following terms: ‘turkey’, ‘poult’, ‘poultry’, ‘broiler’, ‘avian’, ‘flock’, ‘Meleagris’, ‘fowl’, ‘hen’, ‘tom’ or ‘gobble’, in the title or abstract fields. Potential turkey references identified by this script that also reported sampling turkey-origin bacteria in the full text were retained for this review. The data extraction results for these retained turkey references from the initial search, in addition to the data extraction results from the updated search , were then validated against the full text and corrected, if necessary, by a single reviewer, with input from a second reviewer. Exclusion criteria for data extraction of turkey references were the same as the secondary screening criteria .a priori. Descriptive statistics (counts and frequencies) were calculated for factor-level characteristics (details of the study groups and populations in which a factor was present or absent) and outcome-level characteristics  in Excel. For factor-level counts, a factor was deemed a unique combination of the: (1) factor group, (2) comparator group and (3) turkey sub-population . If AMU factors involved the administration of the same antimicrobial or antimicrobial class, they were aggregated together for factor-level counts, even if they involved different doses or routes of administration.The reviewed and corrected data extraction results were then exported from the Access database into Excel. Factors were evaluated for common themes. Themes were not identified The initial and updated searches returned a combined total of 7419 unique citations for primary screening . Of thesCharacteristics of all identified sources of evidence are presented in n = 11; 85%). One reference reported data for breeder turkeys [All references reported factors applied or present at the farm . One stu turkeys  and one  turkeys .n = 27; 75%) were in commercial turkeys and the remainder  were in breeder turkeys. Five themes emerged from the factors identified: (1) AMU policy change (vs. no tylosin use (preventive in-feed)) (vs. no AMU via feed or water) , followed by biosecurity  and management practices . Commercial turkey factors (n = 27) most frequently belonged to AMU themes , followed by biosecurity  and management practices  (n = 9), biosecurity  and management practices  were the most frequent themes.In total, 36 unique factors were identified . Most fay change ; (2) binn-feed)) ; (3) binr water) ; (4) bior water)  and (5) r water) . Almost  7; 26%) . For bren = 3 references), followed by enrofloxacin use, tetracycline (tetracycline and chlortetracycline) use, tylosin use and virginiamycin use (n = 2 references each).Within the binary qualitative and binary quantitative AMU factors in commercial turkeys  and 2, uWithin the binary qualitative AMU theme, the route of administration was either in-feed or unspecified . Some bin = 11) included exposure to other animal populations (n = 5), specific cleaning and disinfection or hygiene practices (n = 4) and water supply (n = 2)  . Within ferences . Three dn = 13), data were reported for AMR in E. coli , Campylobacter species , and Enterococcus species   . None ofn = 9; 69%), fluoroquinolones , macrolides , tetracyclines , penicillins  and other quinolones  (E. coli (n = 9): aminoglycosides  and fluoroquinolones , Campylobacter species (n = 4): macrolides  and Enterococcus species (n = 3): streptogramins . Overall, the bacteria-resistance outcome combinations with the highest frequencies of reporting were E. coli/aminoglycoside , E. coli/fluoroquinolone , E. coli/penicillin , E. coli/tetracycline , Campylobacter species/macrolide , E. coli/other quinolone  and E. coli/cephalosporin   . Within  4; 31%) .Campylobacter species, Enterococcus species and E. coli from turkeys. Identified factors spanned AMU themes, as well as non-AMU themes (biosecurity and management practices). This review also revealed important data gaps; no factors pertaining to S. enterica or to stages in the farm-to-fork pathway other than the farm  were identified, and only one Canadian reference was found.This review identified factors potentially linked with antimicrobial-resistant The final review included 13 references. There was limited depth in the literature, particularly within specific bacteria-resistance outcome combinations. This is consistent with findings from other reviews of factors potentially linked with AMR in livestock , 20. TheThere were substantial data gaps regarding modifiable factors potentially linked with AMR in turkey production. For one, no factors were identified after the farm stage (e.g. abattoir and retail). Interventions after the farm stage may be responsible for the largest reductions in foodborne illness, including antimicrobial-resistant foodborne illness , 53. OthS. enterica. Non-typhoidal salmonellosis is nationally notifiable in Canada [Salmonella Reading have been isolated from Canadian turkey farms, and FoodNet Canada has reported genetic links between human cases of salmonellosis and turkey manure [Salmonella [Salmonella was a substantial gap.Another important data gap was the absence of factors potentially associated with resistance in n Canada . It has n Canada . Also, Sy manure , 57. Givlmonella , 11, theAMU is widely considered the main contributor to AMR , 6 and tLegislative changes related to AMU were identified in one reference: (1) incentive tools, followed by (2) a mandatory ban of the preventive use of critically important antimicrobials , 39. TheBiosecurity and management practices were the most commonly reported themes for breeder turkeys and were also reported in commercial turkeys. This was expected; flock health is particularly important to the breeder industry given its careful genetic selection and role in supplying hatching eggs for turkey meat production . BiosecuSpecific management practices and biosecurity measures of potential relevance to Canadian commercial turkey production were identified in this review, including rearing turkeys in reused litter material, movement to new housing partway through a production cycle, the age of the youngest birds in the flock and disinfection of floors and walls at depopulation. However, several data gaps relevant to Canadian turkey production remain. The Turkey Farmers of Canada On-Farm Food Safety Program has biosecurity and flock care requirements  between consecutive flocks in brooding barns) . These rCertain production practices may lead to the transmission of bacteria, including resistant bacteria, between flocks. In Canada, turkeys may be moved to a grow-out barn after brooding, where dry cleaning is only required yearly , 66. Addvs. dry cleaning vs. disinfection) and bacteria  [Disinfection of barn floors and walls was also a factor identified. Canadian turkey producers may disinfect barn surfaces between consecutive flocks after standard dry cleaning, as disinfection can further reduce the presence of microorganisms important to public health and animal health, including resistant bacteria . The effmonella) .Given the high cost of production practices, including new bedding material, and the environmental challenges posed by the extra waste generated from frequent litter replenishment , more reWhile this review filled an important gap in the literature regarding turkeys and AMR, its approach had limitations. This review only included references reporting AMR in individual bacterial species. Reports of AMR from metagenomics or where only resistance genes were reported  were excluded. Next-generation sequencing technologies may improve the understanding of food systems as complex bacterial ecosystems and are thus expected to assist foodborne AMR surveillance . For exaFurthermore, relevant references may not have been captured on account of the decision to use frequency thresholds in the search strategy (Supplementary Tables S1–S10). These constrained the number of citations captured, to reduce the number of ineligible results and the time necessary to complete relevance screening. As scoping reviews are broad by nature , there iGiven the paucity of turkey-specific studies identified, more research is needed to recommend specific practices to the Canadian turkey industry. It is considered best practice to base policy decisions on the findings of multiple studies examining similar research questions, summarised using systematic review and meta-analysis , 19. DueCampylobacter species, Enterococcus species and E. coli of turkey origin. No factors potentially associated with AMR in Salmonella were identified, and this should be a priority for future work. In addition, only one Canadian reference was found, and no factors pertaining to stages of the farm-to-fork pathway other than the farm were identified.Knowledge syntheses on factors potentially linked with AMR in turkeys are limited, as cattle, chickens and pigs are often the focus of such work , 25–29. This scoping review is an important addition to the body of literature on foodborne AMR. The findings from this review will be incorporated into the larger iAM.AMR project to evaluate potential interventions for reducing the overall occurrence of AMR along the farm-to-fork pathway. Research on the impacts of Canadian industry practices on AMR is necessary to develop a better understanding of AMR in turkeys. Consumption of turkey meat is an important potential source of human exposure to foodborne bacteria, including resistant foodborne bacteria in Canada. The findings from this review (factors reported and data gaps) will play a key role in informing priorities for future research, surveillance and understanding of the epidemiology of AMR in turkey production and associated public health risks.


Molecular subtypes of breast cancer are important references to personalized clinical treatment. For cost and labor savings, only one of the patient’s paraffin blocks is usually selected for subsequent immunohistochemistry (IHC) to obtain molecular subtypes. Inevitable block sampling error is risky due to the tumor heterogeneity and could result in a delay in treatment. Molecular subtype prediction from conventional H&E pathological whole slide images (WSI) using the AI method is useful and critical to assist pathologists to pre-screen proper paraffin block for IHC. It is a challenging task since only WSI-level labels of molecular subtypes from IHC can be obtained without detailed local region information. Gigapixel WSIs are divided into a huge amount of patches to be computationally feasible for deep learning, while with coarse slide-level labels, patch-based methods may suffer from abundant noise patches, such as folds, overstained regions, or non-tumor tissues. A weakly supervised learning framework based on discriminative patch selection and multi-instance learning was proposed for breast cancer molecular subtype prediction from H&E WSIs. Firstly, co-teaching strategy using two networks was adopted to learn molecular subtype representations and filter out some noise patches. Then, a balanced sampling strategy was used to handle the imbalance in subtypes in the dataset. In addition, a noise patch filtering algorithm that used local outlier factor based on cluster centers was proposed to further select discriminative patches. Finally, a loss function integrating local patch with global slide constraint information was used to fine-tune MIL framework on obtained discriminative patches and further improve the prediction performance of molecular subtyping. The experimental results confirmed the effectiveness of the proposed AI method and our models outperformed even senior pathologists, which has the potential to assist pathologists to pre-screen paraffin blocks for IHC in clinic. Breast cancer is intrinsically heterogeneous and has been commonly categorized into molecular subtypes since the late 1990s . MoleculIn clinic, molecular subtype diagnosis usually comes from immunohistochemistry (IHC) . IHC useChanges in gene expression will cause variations in texture in pathological images. Some pathologists have attempted to investigate the statistical relationship between specific gene expression with hematoxylin and eosin (H&E)-stained pathological images . DirectlMolecular subtyping on H&E-stained pathological images is a challenging task since we can only obtain the slide-level label for each molecular subtype without detailed local region information. Even experienced pathologists have difficulty annotating corresponding molecular subtype regions in H&E pathological images . Due to Deep learning is becoming increasingly widely used in computer vision tasks. Most deep learning tasks require a large amount of fine-labeled data for supervised learning, which is time-consuming, especially in medical fields. Weakly supervised learning, for example, has been a hotspot for research on reducing the dependence on labeling data. Benenson et al.  adopted In recent years, multi-instance learning (MIL)  methods Nevertheless, breast cancer molecular subtyping specifically on H&E images has been insufficiently studied. Shamai et al.  used login situ and invasive ductal carcinoma for breast cancer, where pathologists can label tumor regions with different pathological classes, it is impossible to distinguish tumor regions representing different molecular subtypes even for senior pathologists. Although tumor region annotations are useful information for deep networks to learn molecular subtypes, these manual annotations are time-consuming for pathologists. This paper focuses on molecular subtyping with only slide-level labeling instead of detailed tumor region labeling information. The crucial challenge is to eliminate the influence of noise patches and learn expressive features for classifying molecular subtypes.Since the patches cut from each WSI may come from various regions including lesion, benign, or background of the WSI, some research , 26 regaIn this paper, we modeled the patch-based molecular subtype prediction task of pathological slides as a noisy labeling problem in weakly supervised learning. A multi-instance learning framework DPMIL for pathological image molecular subtyping prediction based on discriminative patch filtering was proposed. First, in order to distinguish noise patches, a pre-classification strategy for molecular classification of pathological slides based on co-teaching was presented. This method adopted co-teaching strategy to train two backbone networks and used co-teaching loss function to filter out noise patches to update model parameters. Then, a local outlier factor algorithm was used to reveal the outliers in the feature space for each molecular subtype, and the patches with features close to the cluster center were retained as discriminative patches. Finally, based on the filtered discriminative patches, the pathological slide-level global loss and patch-level local loss were integrated to fine-tune the prediction model for better feature representation of molecular subtypes. The experimental results confirmed the effectiveness of our proposed framework on the molecular subtyping dataset; breast cancer pathological images were provided by Xiangya Hospital. Our AI models outperformed even senior pathologists, which has the potential to assist in pre-screening proper paraffin block of patients for subsequent IHC molecular subtyping in clinic.This paper used breast cancer H&E pathology dataset BCMT (Breast Cancer with Molecular Typing) provided by Xiangya Hospital. All the pathology WSIs used a pyramid storage structure.As We use 4 GeForce GTX 2080 Tis with 11 GB memory to train the network and Python with Pytorch to implement our algorithm. The initial learning rate is 0.1 and the poly learning rate policy with the power of 0.9 is employed. The minibatch size is set as 32.This paper proposes a breast cancer molecular subtype prediction framework based on multi-instance learning and discriminative patch filtering. The pipeline of our framework is illustrated in Firstly, patches from H&E WSIs are extracted to train a molecular subtype classifier. Co-teaching  between In multi-instance learning framework, each patch is usually assigned the same label as WSI it belongs to –31, whilK follows , which is a classic density-based algorithm . The maiWe perform LOF for each subtype of molecular features and regard patches that do not belong to a specific cluster of molecular subtype as noise patches.The above selected discriminative patches are further used to improve feature representation of molecular subtypes based on multi-instance learning framework (MIL). MIL regards the WSI as a bag containing a number of patches. These patches are considered as instances, and their predictions are aggregated to obtain a bag-level prediction. ResNet-50 is also adopted as a backbone to train the MIL classification model. We initialize the MIL model with the model that performs better in co-teaching and use discriminative patches for fine-tuning.WSIL is defined as Formula 1, where WSIL represents the slide-level loss function of the ith pathological image defined as the cross-entropy function , the National Natural Science Foundation of China (81972490) and Natural Science Foundation of Hunan Province (2019JJ50781).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Background: The available evidence suggests that amoxicillin is often associated with the occurrence of Kounis syndrome (KS). The purpose of this study is to explore the clinical characteristics of KS induced by amoxicillin.Methods: We searched for case reports of amoxicillin-induced KS through Chinese and English databases from 1972 to May 2022.Results: A total of 33 patients with KS were included, including 16 patients (48.5%) receiving amoxicillin treatment and 17 patients (51.5%) receiving amoxicillin-clavulanate. The median age was 58 years (range 13–82), 75.8% were from Europe and 81.8% were male. Nearly 70% of KS patients develop symptoms within 30 min after administration. Chest pain (63.6%) and allergic reaction (75.8%) were the most common clinical manifestations. Diagnostic evaluation revealed elevated troponin (72.7%), ST-segment elevation (81.2%) and coronary artery stenosis with thrombosis (53.6%). Thirty-two (97.0%) patients recovered completely after discontinuation of amoxicillin and treatments such as steroids and antihistamines.Conclusion: KS is a rare adverse reaction of amoxicillin. Amoxicillin-induced KS should be considered when chest pain accompanied by allergic symptoms, electrocardiogram changes and or elevated levels of myocardial injury markers. Therapeutic management of KS requires simultaneous treatment of cardiac and allergic symptoms. Epinephrine should be used with caution in patients with suspected KS. Kounis syndrome (KS) is an acute coronary syndrome caused by an allergic reaction, and first reported by Kounis and Zavras in 1991 . PatientA variety of reasons have been found to induce KS, including many drugs, diseases, food, environmental exposure or certain other conditions . Among tWe searched the literature related to amoxicillin-induced KS by searching Chinese databases , Chinese VIP) and English databases  from 1972 to May 2022. The search combined subject and free words such as amoxicillin, amoxicillin-clavulanate, Kounis syndrome, antibiotics, allergic reactions, β-lactams, thrombosis, myocardial infarction, acute coronary syndrome, coronary spasm, hypersensitivity.Inclusion criteria: case report and case analysis of KS induced by amoxicillin. The clinical data is relatively complete, including the amoxicillin application, clinical manifestations, laboratory examinations, treatment and prognosis etc. Exclusion criteria: duplicate literature, reviews, mechanistic studies, animal studies and articles for which the full text was not available.Two researchers independently conducted a preliminary screening of the literature according to the inclusion and exclusion criteria, and then the group discussed the included literature. We extract the following information of patients: region, gender, age, medical history, drug combination, amoxicillin application, indication, symptom onset time, clinical manifestations, laboratory examinations, imaging examinations, treatment and prognosis by using self-designed data extraction table.Statistical analyses were performed using SPSS 22.0 . Continuous data is represented by median value with ranges, counting data is represented by number of cases and percentage (%).As shown in In these patients, 16 patients (48.5%) received amoxicillin and 17 patients (51.5%) received amoxicillin-clavulanate . The daiThe clinical symptoms of 33 KS patients are summarized in The laboratory tests of 33 KS patients are summarized in The imaging examinations of 33 KS patients are summarized in The treatment and prognosis of the 33 KS patients are summarized in Sixteen patients (48.5%) belonged to type I KS variant, 10 patients (30.3%) belonged to type II KS variant, 2 patients (6.1%) belonged to type III KS variant. The KS variant could not be identified in the remaining 5 patients (15.2%).KS is an allergic acute coronary syndrome that can occur at any age, but the most commonly affected age group is 40–70 years (68%) of male patients (74.3%). Patients with a history of allergies, hypertension, smoking, diabetes, and hyperlipidemia are more likely to be affected . Among tIt is currently believed that the occurrence of KS is caused by allergic reactions in people with allergies after exposure to specific antigens. The main inflammatory cells that are involved in the development of KS are mast cells that interact with macrophages and T-lymphocytes . InfiltrAt present, the guideline for the treatment of KS have not been established, and the treatment recommendations are mainly derived from the experience summary of case reports. The treatment of KS should consider two aspects of acute coronary syndrome (ACS) and allergic reaction. Patients with ACS should be treated according to the ACS guidelines. Anti-allergic treatment often has a better effect in patients with type I KS variant, while patients with type II variant and III KS variant need to treat acute coronary syndromes while being anti-allergic . CorticoKS has a good prognosis and can fully recover with appropriate treatment in most patients. Our research showed that amoxicillin-induced KS may have serious complications, such as cardiac arrest in 12.1% of patients and death in 3% of patients.In conclusion, KS is a rare adverse reaction of amoxicillin. Amoxicillin-induced KS should be considered when chest pain accompanied by allergic symptoms, ECG changes and or elevated levels of myocardial injury markers.


To develop biodegradable, sustainable, and environment-friendly functional food-packaging materials, gelatin/zein/glucose nanofibers were fabricated through air-assisted electrospinning and then crosslinked by the Maillard reaction under mild conditions (60 °C and 50% relative humidity) in this study. Compared to traditional electrospinning, air-assisted electrospinning increased the yield of nanofibers by 10 times, and the average diameter from 263 nm to 664 nm, while the airflow facilitated uniform and smooth nanofiber formation. During the Maillard reaction in 0–5 days, the gelatin/zein/glucose showed no morphology change. Fourier transform infrared spectra analysis indicated that gelatin interacted with zein through hydrogen bonding and the occurrence of the Maillard reaction among the protein and glucose molecules. After four days of Maillard reaction, the nanofibers presented higher thermal stability, the most hydrophobic surface (water contact angle: 133.6°), and stiffer network structure . Overall, Maillard-reaction-crosslinked gelatin/zein/glucose nanofibers showed favorable physical properties, which suggests their potential for application in food-active packaging. Functionalized electrospun nanofibers possess multiple attributes, e.g., moderate preparation conditions, high porosities, high surface/volume ratio, outstanding mechanical properties and acceptable morphologies, and so on, which provide excellent prospects for application in the field of food. Currently, many studies mainly focus on antioxidant packages , antimicGelatin is a water-soluble protein derived from animal tissue, while zein is a hydrophobic storage protein from corn that is highly biocompatible and exhibits film-forming properties ,8. Thus The Maillard reaction, which involves grafting the amino groups of a protein with the carbonyl moieties of a reducing sugar, has been reported to offer a green chemical crosslinking strategy . Kchaou In addition, the low yield afforded by traditional electrospinning is another obstacle to the use of nanofibers in the food industry. Air-assisted electrospinning technology is considered to have excellent application prospects, and adding airflow forces based on traditional electrospinning technology has been shown to improve performance and significantly increase yield ,19. AminThis study aimed to fabricate gelatin/zein/glucose nanofibers through air-assisted electrospinning, with subsequent crosslinking by the Maillard reaction under mild conditions. We hypothesized that gelatin/zein nanofiber films with physical properties suitable for food applications will be produced by the Maillard reaction under mild conditions. Scanning electron microscopy (SEM) was used to evaluate the microscopic morphology and diameter distribution of nanofibers. To interpret changes in macroscopic performance such as the water contact angle (WCA), mechanical property, water vapor permeability (WVP), and water stability, the interaction between protein and glucose was studied using Fourier transform infrared (FTIR), X-ray photoelectron spectroscopy (XPS), and thermal analysis techniques.Gelatin , glucose, and acetic acid were purchased from Aladdin Reagent Database Inc . Zein  was purchased from Sigma Aldrich .w/v) and zein (15% w/v) were dissolved in 80% aqueous acetic acid. Glucose (5% w/v) was then added to the solution in accordance with our previous report [Gelatin  at 60 °C and 50% relative humidity (RH) for 0, 1, 2, 3, 4, and 5 days.Vacuum gold spraying was applied to treat the fibers, and SEM was utilized to evaluate their microscopic morphology . Using Nano Measurer 1.2, the diameters of forty randomly selected fibers were determined from the SEM images, and the related diameter distribution was estimated and fitted.The colorimetric a* (redness) and b* (yellowness) values of the fiber films were measured using a chroma meter .−1; resolution: 2 cm−1; number of scans: 32). Prior to each scan, air was used as the background and subtracted. The infrared spectra of nanofibers crosslinked for 0–5 days were compared quantitatively by calculating the relative absorbance changes (∆RA). The absorbance of the peak was used divided by the sum of the absorbance of all the peaks detected in the spectrum [Using a Fourier transform infrared (FTIR) spectrometer, infrared spectra of a tiny sample taken from the nanofibers were captured  and thermogravimetric analysis (TGA) , the thermal characteristics of the nanofibers were identified. Each sample was precisely weighed at 6–10 mg and sealed in an aluminum crucible. The sample was then heated under dry NXPS  was used to determine the chemical state and composition of the nanofiber surface. The spectrum survey scans were obtained over the 0–1350 eV binding energy range at a detector pass energy of 100 eV and high-resolution spectra were recorded for the C1s region at pass energy of 50 eV. The spectra were processed using Advantage software  and OriginPro 2022b  [The WCA of the nanofibers was carried out using a tensiometer . A droplet of liquid (3 µL) was deposited on the film surface (20 × 40 × 0.2 mm) with a precision syringe, and the contact angles were measured at 5 s after the drop contacted the nanofibers.Using a DR-508A  computer tensile testing machine, the tensile strength (TS), elastic modulus (EM), and elongation at break (EB) were calculated. The nanofibers, which were around 0.1 mm thick, were sliced into 5 × 1.5 cm pieces and put in a fixture, after which the thickness, width, and intercept of the samples were precisely measured, entered into the software, and then subjected to a tensile load of 5 N and a tensile rate of 5 mm/min. Each sample was measured five times. Following are the calculations for the TS, EM, and EB :(1)Tensi2); L is the thickness (cm); ∆P is the rated vapor pressure differential (Pa) ; Ws/t is the linear regression of weight over time (g/s).The lip of a 10 mL water-permeable cup was sealed with a sheet of uniformly thick, 6 cm diameter nanofiber film, and the cup was then placed in a desiccator. Hourly weights and records were kept for all samples. This process was repeated three times over 6 h. WVP was calculated as :(4)WVP(gNanofibers (5 mg) were soaked in water for two months and then vacuum freeze-dried . The microscopic morphologies of the fibers were observed by SEM.p-value < 0.05 was considered statistically significant.The quantitative data were statistically analyzed and expressed as means ± standard deviations. The statistical significance of mean values between multiple treatment groups was accessed by one-way analysis of variance (ANOVA) with Tukey’s tests using the OriginPro 2022b . A SEM images of gelatin/zein/glucose nanofibers fabricated by traditional and air-assisted electrospinning are presented in SEM images of electrospun gelatin/zein/glucose films obtained after the nanofibers had been subjected to the Maillard reaction at 60 °C for 24 h at various RH are presented in −1 was assigned to the O–H and N–H stretching vibrations that may be associated with hydrogen bonding interactions between gelatin and zein [−1 was attributed to the –CH2 stretching vibration of aliphatic groups [−1, 1541 cm−1, and 1241 cm−1 were ascribed to the C=O stretching vibrations of the amide I, the C–N stretching vibration of amide II, and C–N stretching and N–H bending combination of amide III, respectively [−1 and 1031 cm−1 resulted from the C–O stretching of glucose [and zein . The banc groups . The absectively . The pea glucose .−1) and amide II (1541 cm−1) were significantly enhanced, and the peaks (1080 cm−1 and 1031 cm−1) representing the C-O stretching of glucose were reduced considerably, to prove the occurrence of glycation.Relative absorbance of the peaks after Maillard reaction for 0~5 days is listed in The DSC and TGA curves of the films after Maillard reaction for 0~5 days are presented in As shown in The WCA of the nanofibers after Maillard reaction for 0~5 days is presented in The elemental compositions of the nanofiber surfaces were determined by XPS, the results of which are shown in The three characteristic peaks observed at 284.7, 286.2, and 288.0 eV in the high-resolution C1s spectra in The water vapor permeability (WVP) of nanofibers after Maillard reaction for 0~5 days is shown in In this study, gelatin/zein nanofibers were successfully fabricated using air-assisted electrospinning, and then glycated with glucose by the Maillard reaction. The significantly higher yield of nanofibers obtained by air-assisted electrospinning highlights its potential as an alternative technology to the traditional electrospinning. The crosslinked nanofiber films exhibited beneficial mechanical strength, hydrophobicity, structural stability, and low color browning after the Maillard reaction under mild conditions, which indicates that natural matrix nanofibers can be modified in a green and cost-effective manner. Therefore, the mild Maillard reaction conditions reported herein offer a promising nanofiber modification method for temperature-sensitive bioactive delivery for the food and biomedical industries.


Job losses have been widespread, and individuals have experienced significant restrictions on their usual activities, including extended isolation from family and friends. While studies suggest population mental health worsened from before the pandemic, not all individuals appear to have experienced poorer mental health. This raises the question of To understand the coping strategies individuals employed during the COVID-19 pandemic, we used structural topic modelling, a text mining technique, to extract themes from free-text data on coping from over 11,000 UK adults, collected between 14 October and 26 November 2020.We identified 16 topics. The most discussed coping strategy was ‘thinking positively’ and involved themes of gratefulness and positivity. Other strategies included engaging in activities and hobbies , keeping routines, and focusing on one day at a time. Some participants reported more avoidant coping strategies, such as drinking alcohol and binge eating. Coping strategies varied by respondent characteristics including age, personality traits and sociodemographic characteristics and some coping strategies, such as engaging in creative activities, were associated with more positive lockdown experiences.A variety of coping strategies were employed by individuals during the COVID-19 pandemic. The coping strategy an individual adopted was related to their overall lockdown experiences. This may be useful for helping individuals prepare for future lockdowns or other events resulting in self-isolation. While sHow people cope is an important factor underlying the relationship between experiencing stressors and subsequent mental health. Coping is generally defined as the cognitive and behavioural efforts that are used to manage stress . There iA number of studies have examined coping during the COVID-19 pandemic, using quantitative , qualitaThere has also been a methodological challenge with the studies on coping during COVID-19 carried out so far. Existing quantitative studies have typically relied on closed-form responses to survey items . An issue with this approach is that responses are restricted to those that the researcher has thought of in advance – an issue that is particularly salient given the novelty of the COVID-19 pandemic. While qualitative approaches allow for more flexibility in responses, the small sample sizes typical of qualitative studies restrict the questions that can be asked of the data – specifically, those that statistically relate individual characteristics and circumstances to topics raised.Text-mining methods offer the benefits of quantitative and qualitative approaches, enabling the extraction of themes from large-scale free-text data that can be summarised numerically and related to participant characteristics  using standard statistical methods. To our knowledge, two studies have used text-mining methods to analyse free-text survey data on coping during the COVID-19 pandemic in the United Kingdom . Both fovia our first strategy, including (i) individuals from a low-income background, (ii) individuals with no or few educational qualifications, and (iii) individuals who were unemployed. Third, the study was promoted via partnerships with third sector organisations to vulnerable groups, including adults with pre-existing mental health conditions, older adults, carers, and people experiencing domestic violence or abuse. Full details on sampling, recruitment, data collection, data cleaning and sample demographics are available at https://doi.org/10.17605/OSF.IO/JM8RA. The study was approved by the UCL Research Ethics Committee (12467/005) and all participants gave informed consent.We used data from the COVID-19 Social Study; a large panel study of the psychological and social experiences of over 70,000 adults (aged 18+) in the United Kingdom during the COVID-19 pandemic. The study commenced on 21 March 2020 and involved online weekly data collection for 22 weeks with monthly data collection thereafter. The study is not a random sample and therefore is not representative of the United Kingdom population, but it does contain a heterogeneous set of individuals. Participants were recruited in three ways. First, convenience sampling was used, including promoting the study through existing networks and mailing lists , print and digital media coverage, and social media. Second, more targeted recruitment was undertaken focusing on groups who were anticipated to be less likely to take part in the research What have been your methods for coping during the pandemic so far and which have been the most or least helpful? . To predict topic proportions, we included variables for age, sex, ethnicity, country of residence, education level, living arrangement, keyworker status, self-isolation status, diagnosed psychiatric condition, long-term physical health conditions, and Big-5 personality traits.Country of residence , sex , ethnicity , age . Free-text responses were cleaned using an iterative process. The main steps were as follows. Popular hyphenated words were collapsed into non-hyphenated form and spaces were removed between words that could have been hyphenated . Punctuation mistakes  were replaced with whitespace unless the full stop denoted an initialism or a URL. Full stops were removed between initialisms – e.g., U.K. became UK – and “www.” was removed from URLs. Responses were tokenized into lower-case unigram form and “stop” words (common words such as “the” and “and”) were removed. Stop words were identified with the onix, SMART, and snowball dictionaries , excludin = 113), so we used complete case data.We performed several quantitative analyses. First, as not all participants chose to provide a response, we ran a logistic regression model to explore the predictors of providing a free-text response. We used the variables defined above as predictor variables . Second, we used STM, implemented with the stm R package , to extrn = 2,203).We ran STM models from 2 to 30 topics and selected the final models based on visual inspection of the semantic coherence and exclusivity of the topics and close reading of exemplar documents representative of each topic (documents with highest proportion of text estimated as belonging to a given topic). Semantic coherence measures the degree to which high probability words within a topic co-occur, while exclusivity measures the extent to that a topic’s high probability words have low probability for other topics. After selecting a final model, we carried out three further analyses. First, we decided upon narrative descriptions for the topics based on high probability words, high “FREX” words (a weighted measure of word frequency and exclusivity), and exemplar texts. Second, we ran multiply-adjusted linear regression models estimating whether topic proportions were related to author characteristics defined above (again categorising age into four groups to aid interpretability). For comparability with categorical variables, Big-5 personality trait variables were scaled such that a 1-unit change was equal to a 2 SD difference .  included individuals who had tried to see the positives in the situation, to remember that others were in relatively worse situations, and recognise that the pandemic would pass. Topic 6  similarly, related to a general cognitive coping strategy, including text on individuals taking each day as it came and imposing structure on their time. This topic overlapped with Topic 12 , which related to people keeping routines, particularly with exercise. Similarly, Topic 10  related to participants filling their time (“keeping busy”) in generally non-specific ways.Most other topics related to spending time on specific activities. Topic 3  related to individuals engaging in arts, hobbies, or crafts as a way of coping. Topic 5  included text from participants who reported spending their time listening to music and radio or watching TV and films. Topic 4  related to individuals who had used the opportunity to take long walks and get into nature, while Topic 15  included text from individuals who found exercise had a positive effect. Topic 8  and Topic 11  including responses on keeping in contact with family, friends and colleagues, the latter referring to the use of online technologies in particular. Topic 9  referred to individuals spending time gardening or completing “odd jobs” at home. Topic 14  included participants spending time on activities online, including classes, courses, and group sessions (such as singing groups) as well as functional online activities such as ordering supermarket deliveries.Amongst the remaining topics, Topic 2  included individuals who reported self-harming or increasing alcohol consumption or comfort eating, though the latter two were reported in several cases as improving mood (at least in the short term). Topic 16  meanwhile included text on individuals actively avoiding coverage on COVID-19 as a coping strategy. Topic 7  referred specifically to attempts to reduce risk by following guidelines . Finally, Topic 13  surfaced exemplar texts that did not contain a clear, consistent theme.p-values . This suggests that our models extracted consistent and meaningful themes. While structural topic models are novel in the coping literature, our results show that such models can complement and bridge qualitative and quantitative approaches, providing insights not easily attained with either approach on its own. A further strength of this study was that we used data from 7 to 8 months after the first lockdown, allowing for an assessment of coping strategies across an extended period of the pandemic.Nevertheless, this study had several limitations. Not all of the topics identified a single theme consistently and associations with participant characteristics could be driven by idiosyncratic texts. Our sample, though heterogeneous, was not representative of the United Kingdom population. Respondents to the free-text question were also biased towards the more highly educated. This may have generated bias in the topic regression results. While it is plausible that participants discussed coping strategies that they deemed most important, participants may have employed multiple coping strategies and not written about them all. Further, across the long timespan of the pandemic, individuals may have adopted different strategies at different points. Responses may have been biased towards those salient at the time . Moreover, individuals may not interpret or be aware of a behaviour as a coping strategy, though it has that effect – for instance, increasing consumption of alcohol or fatty or sugary foods. A final limitation was that, while we included a wide set of predictors in our models, many relevant factors were unobserved. Associations may be biased by unobserved confounding.Sixteen different coping strategies employed by adults in the United Kingdom during the COVID-19 pandemic were identified through text-mining participant free test responses to the COVID-19 Social Study. Some strategies reported were more cognitive (or “antecedent-focused”), either based around attentional deployment (both focusing attention onto the pandemic by focusing on following the rules or distracting oneself from events by avoiding the news), problem solving  or cognitive change (e.g. trying to think more positively about things) . Others liam.wright@ucl.ac.uk.The datasets presented in this article are not readily available due to stipulations made by the ethics committee. Requests to access the datasets should be directed to LW, The studies involving human participants were reviewed and approved by UCL Research Ethics Committee (12467/005). The patients/participants provided their written informed consent to participate in this study.All authors conceived and designed the study. LW curated the data and conducted the data analysis. LW and MF agreed on narrative titles for the topics and wrote the first draft. All authors provided critical revisions, read, and approved the submitted manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Arachidonic Acid (AA) is the precursor of cerebrovascular active substances in the human body, and its metabolites are closely associated with the pathogenesis of cerebrovascular diseases. In recent years, the cytochrome P450 (CYP) metabolic pathway of AA has become a research hotspot. Furthermore, the CYP metabolic pathway of AA is regulated by soluble epoxide hydrolase (sEH). 1-trifluoromethoxyphenyl-3(1-propionylpiperidin-4-yl) urea (TPPU) is a novel sEH inhibitor that exerts cerebrovascular protective activity. This article reviews the mechanism of TPPU's protective effect on ischemic stroke disease. Ischemic stroke is a serious condition that endangers human health. The prevalence and incidence of stroke in China is rising faster than in other countries due to an aging population, the continued high prevalence of risk factors such as hypertension and diabetes and irregular management. Studies have shown that the prevalence of stroke among Chinese residents aged ≥40 years is 2.58%, or ~17.5 million people. The prevalence of stroke in the adult population (≥18 years) is ~1.29%, with significantly higher increases in the male population and in urban areas , and overall China is facing the greatest stroke challenge in the world (Arachidonic acid (AA) is the precursor of cerebrovascular active substances in the human heart, and its metabolites are closely associated with the pathogenesis of cerebrovascular diseases . In rece2)  –9. Anothreatment . 1-triflin vivo, resulting in the failure of clinical translation of traditional sEH inhibitors.Neuroprotection against sEH has been a hot topic in recent years. Although EPHX2 gene knockout has a protective effect on experimental cerebral ischemia, it is still distant from clinical prevention and treatment of ischemic stroke. Therefore, sEH inhibitors R&D has attracted much attention. In 2005, the she inhibitor 12-(3-adamantan-1-yl-ureido)-dodecanoic acid (AUDA) was applied to the cerebral ischemic nerve. In protective experimental studies, it has been confirmed that AUDA has a protective effect on cerebral ischemia . FurtherTPPU is a novel sEH inhibitor synthesized in 2012 by Professor Bruce from the Molecular Bioscience Center of the University of California Veterinary Medicine . Bruce Hin vivo reduced atherosclerotic lesions of EDN mice . The main pathogenic changes in the early stages of cerebral ischemia are increased BBB permeability and damage of tight junction protein, which causes brain edema, directly or indirectly leading to neurological function damage. Hence, preventing ischemic stroke requires protecting the BBB, inhibiting increased permeability, and alleviating cerebral edema . Yi et aoverused . Furtheroverused .The decrease in perfusion volume after cerebral infarction is the primary reason for the aggravation of neurological function. Low perfusion can not only stimulate the intensification of the local inflammatory response but also result in the activation of excessive oxygen free radicals, causing the aggravation of the disease . Hao et Apoptosis is one of the primary reasons for brain injury after ischemic stroke, and oxidative stress is the main pathway of apoptosis. Studies have shown that TPPU can reduce the production of reactive oxygen species (ROS) after ischemic stroke, increase the expression of Bcl-2 protein, and decrease the expression of Bax protein, implying that TPPU can inhibit cell apoptosis after ischemic stroke . ROS proin vitro and poor or no activity in vivo to the current sEH inhibitors with nanomolar activity in vitro and remarkable pharmacokinetic properties in vivo (Due to the widespread existence of sEH in the human body, its metabolic pathway is involved in many diseases, including ischemic stroke, hypertension, heart disease, kidney disease, etc. In theory, the development of drugs that inhibit sEH would seem to reverse or treat the disease. Human drug exploration for sEH has never stopped. From the early discovery of epoxide sEH inhibitors to the design and synthesis of the third generation of urea human sEH inhibitors, the research on human sEH inhibitors has experienced more than 30 years of development. sEH inhibitors have progressed from the initial sEH inhibitors with only micromolar inhibitory activity  in vivo .in vivo. It has been used in basic research in various cardiovascular and cerebrovascular diseases. It can play a protective role through a variety of potential mechanisms. However, it has not yet entered the clinical research stage for cerebrovascular diseases. With the development of pharmacology, the effects and mechanisms of TPPU are constantly being discovered and clarified, further proving its rationality and effectiveness as a treatment for ischemic stroke. As the research progresses, TPPU and its derivatives could be novel drugs for treating ischemic stroke-related diseases.TPPU is a novel sEH with good activity In conclusion, TPPU can intervene in ischemic stroke in various ways and may be one of the new targets for treating acute ischemic stroke. However, the treatment of acute ischemic stroke with TPPU is still in the exploratory stage, no clinical research has been carried out yet, and more trials are needed to confirm its potential.The author confirms being the sole contributor of this work and has approved it for publication.