N = 4,376), to understand public responses to the Lorry Deaths incident. We find that Twitter users' posts were organized into two themes regarding attitudes toward immigrants: (1) migration narratives, stereotypes, and victim identities, and (2) border control. Within each theme, both pro- and anti-immigration attitudes were expressed. Pro-immigration posts reflected counter-narratives that challenged the mainstream media's coverage of the incident and critiqued the militarization of borders and the criminalization of immigration. Anti-immigration posts ranged from reproducing stereotypes about Vietnamese immigrants to explicitly blaming the victims themselves or their families for the deaths. This study demonstrates the uses and limitations of using Twitter for public opinion research by offering a nuanced analysis of how pro-and anti-immigration attitudes are discussed in response to a tragic event. Our research also contributes to a growing literature on public opinion about an often-forgotten immigrant group in the UK, the Vietnamese.In this article, we analyze anti- and pro-immigrant attitudes expressed following the Essex Lorry Deaths tragedy in October 2019 in Britain, in which 39 Vietnamese immigrants died in a sealed lorry truck on their way to their destination. We apply Structural Topic Modeling, an automated text analysis method, to a Twitter dataset ( Humanitarian tragedies tend to receive immediate attention and responses by the public on social media. Computational science research on Twitter responses to such events allows us to examine the public's responses in real time. Such research can access the immediate responses to social issues related to a tragic event, such as immigration, border control policies, white nationalist attacks, and more. While much of the research on public attitudes toward immigrants and border control involves analyzing survey data, the recent emergence of Twitter text data and appropriate methods to study social media data have provided new ways to investigate these topics. Social media platforms enable the public to express their attitudes about immigration issues, to make sense of tragic events, and to express collective trauma  to obtain the tweets pertaining to the Essex Lorry Death incident. We collected all messages that included the following hashtags #Lorrydeaths #Lorrydeath, #Essex, #Containerdeath, #Grays, #Vietnamese, #HumanTrafficking, #39deaths, #moRobinson. These hashtags were used during the 2 weeks immediately after the event took place. Specifically, we retrieved unique tweets created between October 23, 2019, and November 6, 2019, which included at least one of those hashtags. Others who may have participated in the public conversations but did not tag themselves by using the hashtags, would not have been captured by the algorithm. In total, we collected 35,158 unique tweets. Our corpus excluded retweets. Furthermore, non-unique tweets, or repeated tweets for different hashtags are counted only once. We also removed non-English tweets. In our original corpus, tweets from other languages such as Vietnamese also appeared. By manually going through a sample of those Vietnamese tweets, we recognized that they came from Vietnam based on their content. We thus removed those, and tweets in other languages such as Dutch, German, and French. Finally, we manually examined the remaining tweets to exclude irrelevant ones—for example, those that describe weather or advertise for local businesses. After applying all the filter procedures, the final dataset includes 4,376 unique tweets by 2,596 different users.By narrowing the tweets to those in English only we focus on the discourses that this event brought up, specifically as it is relevant to discussions about Vietnamese immigrants, and undocumented immigration in the UK more generallyWe use structural topic modeling (STM) by Roberts et al. , an autoTopic models are unsupervised machine learning models, which automatically discover latent topics from text documents. A topic model presents the researcher with two crucial pieces of information: the distribution of topics within the corpus, and distribution of words per topic. In these models, a topic can be understood as a set of words representing interpretable themes, and documents are represented as a mixture of these topics. For each document, proportions across all topics sum up to 100%. For example, after fitting a model, a document might be captured by a combination of three topics: “border control” with a proportion of 70%, “human trafficking” with 20%, and other topics with 10%. We chose STM over other topic modeling methods such as Latent Dirichlet Allocation (LDA) because STM allows for variability of “word use within a topic” by a covariate  in the final dataset on the day of the tragic event and the two subsequent weeks. Most of the tweets were posted within the first 6 days of the event . In contrast, between October 29 and November 6, only 869 posts, or 19.86% of the dataset's tweets, were posted. This dwindling in the number of tweets after the first few days of the event aligns with “the overall temporal pattern of information diffusion on Twitter”  of authoritarian regimes and blames such regimes as the “root cause” for such tragedies. Here again, the blame for the Lorry deaths is put on a developing country instead of contextualizing such tragedy as part of a global migration system tied to colonialism and border control. The Twitter user cites CNN and Twitter as a way to provide others with news updates. Twitter users are therefore responding to the news media discourse and sharing their thoughts on it. One Twitter users critiqued the anti-China idea that the country has an authoritarian regime and challenged the narrative of Chinese backwardness:When the victims were identified as coming from Vietnam, a poorer country than China, and specifically from the northern central area of Vietnam, one of the poorest regions in the country, the narrative about migration motives changed. The narrative switched from running away from a repressive regime, to running toward economic opportunities. Many attributed the victim's reason for migration to poverty.tweet 11 [Topic 1]: Vietnam is a booming economy but not everyone is benefitting from it. It has a huge surplus of labor. While the country has achieved reductions in #poverty, it is unequal across groups & regions. #HumanTrafficking #Vietnamesetweet 12 [Topic 11]: Migratory pressures are encouraging millions to make long and dangerous journeys to escape suffering and poverty and to seek safer and better lives in countries like #Britain #Essex #Belgium #NorthernIreland #Holyheadtweet 13 [Topic 18]: Cannabis factories in the UK sustained by human trafficking children; teenagers kidnapped from Vietnam; exploited as slaves.tweet 14: [Topic 18]: Vietnamese victims of trafficking may end up working in nail bars, so what's the solution? We told it's time for labor inspection agencies to have proper funding so it can do its job in protecting workers from abuse.tweet 15 [Topic 18]: Vietnamese victims of human trafficking may end up being underpaid and overworked in nail bars. Many would say we need an ethical stamp or similar – that's not the right solution.Others discussed the labor conditions that Vietnamese migrants would find in the UK, which were based on exploitative practices ranging from underpaid jobs in “nail bars” to being “exploited as slaves” in “cannabis factories”:tweet 16 [Topic 18]: The former UK Antislavery Commissioner has criticized the UK approach to tackling Human Trafficking and Modern slavery, stating the need for greater investigative resources and a stronger focus on tackling the Organized Crime networks involved.tweet 17 [Topic 18]: The deaths of 39 migrants found in a truck in #Essex, UK has put the spotlight on trafficking, slavery in the nail bar sector, a common destination for trafficked Vietnamese.In these tweets, Twitter users critique the exploitative practices of the human trafficking cycle between the UK and Vietnam. Here, users charge several actors for sustaining this cycle: the human traffickers, UK cannabis factories, and UK labor inspection agencies. A related discussion concerned human trafficking as a cause for the deaths and exploitation of immigrants:These discussions suggest a search for those responsible for the Essex Lorry Deaths among the human traffickers and the failures of the UK government. One of the demands is to increase interventions into “the organized crime networks involved” (tweet 16). This demonstrates a desire for increased criminalization of human trafficking and organized crime. It demonstrates some trust that the UK government would address these crimes. However, these critiques do not address more structural issues such as UK's immigration policies and can therefore still be read as anti-immigration; these tweets do not actually suggest more humane and welcoming immigration policies, which would reduce the risks involved in immigrating in irregular ways.Furthermore, although these tweets critique exploitative labor practices and the violence of human trafficking, they also reify two stereotypes about Vietnamese immigrants. First, they rely on the stereotype that Vietnamese immigrants in the UK immigrate via human trafficking. In fact, there are four groups of Vietnamese migrants to the UK: “refugees from the former South Vietnam, refugees from the former North Vietnam, new undocumented economic migrants from northern and central Vietnam, and international students” Barber, , p. 10. tweet 18 [Topic 11]: Are none of the networks asking WHY these families sent their sons/daughters here for £30,000 illegally, when for a fraction of that they could have visited here, met with colleges, got advice on visas, looked at student/work options, made useful contacts? #Purfleettweet 19 [Topic 19]: Why can't these people, if their families have access to that kind of money, come here on a visit, do some research, make contacts, ask about work placements or higher education, apply for a visa? #Purfleettweet 20 [Topic 19]: If people are texting their families from inside a container & have paid smugglers 30,000 & the families are aware what is happening then who is to blame?Twitter users also blamed the victims' families for the incident:Each of these Twitter users saw the families as responsible for their family members' deaths. They justified such statements by highlighting the large sum of money  that human traffickers demanded for smuggling people to Europe. The tweets demonstrate a lack of understanding of the reasons why people migrate and the mechanisms of international migration. The immigrants who died in the lorry truck were driven to escape the poorest region in Vietnam. On a structural level, this is consistent with uneven development and globalization where core economies like the UK have the most capital and developing countries like Vietnam supply resources and excess labor leading to economic migration from developing countries to the UK tweet 33 [Topic 12]: Honestly, I hope the media are held to account if the Lorry Driver in the Lorry deaths case is found to be innocent. Naming and showing photo of suspect so early on pre-court hearing is disgraceful and unfair. Should he be found innocent I hope he takes them to the cleaners.This tweet similarly defends the driver, arguing that he is innocent until proven guilty. Furthermore, the Twitter user criticizes the role of the media in spreading photos of the suspect so early on, thus leading to his trial in the court of public opinion before the real court hearing.tweet 34 [Topic 6]: Organized Chinese criminals called snakehead gangs are well known for people smuggling, prompting some to speculate that they may have a hand in the deaths of 39 people found in a refrigerated lorry trailer in Essex.tweet 35 [Topic 20]: Snakehead travel agents in China are transporting people to Europe for 6-26k to work in black market syndicates, according to organized crime expert Dan Silverstone.Other Twitter users accused organized Chinese criminals of smuggling the people who died:These two tweets try to justify their accusations by citing an expert (as in tweet 35). The accusations also evoke anti-Chinese stereotypes and may be related to the initial suspicion by the Western media that the victims were Chinese, as discussed above.tweet 36 [topic 11]: I have zero sympathy toward 39 illegal aliens infiltrating our borders and potentially at risk of putting more native folks on the streets homeless. Draining our systems. #lorrycontainer #gmb #realist.tweet 37 [topic 11]: Prevention is better than cure! Remove the incentives for illegal immigrants! Welfare reform: immediate removal of illegals would dramatically reduce the numbers trying to enter illegally!The incident also stirred up more general xenophobia among a smaller but still vocal group of tweeters. Concerns ranged from fear of crime to competition for jobs and social services, sometimes called “welfare chauvinism” . Other anti-immigrant attitude was shared regarding resources:These Twitter users argue that immigrants could not be poor because they could afford “30 grand” to come to the UK. Tweet 39 claims that “genuine refugees” would not be able to afford such an expensive passage. There are two tropes implied in this argument: the undeserving immigrant, and victim blaming. The language around authenticity constructs migrants as those forced to migrate while others do so voluntarily (or unnecessarily). In line with Flores Morales and Farago  study onThis study's contributions are three-fold: (1) it demonstrates the utility of Twitter for public opinion research, as well as the value of using computational social science to study social media; (2) it offers a more nuanced understanding of how pro-and anti-immigration attitudes are expressed in response to a tragic event; and (3) it expands the limited literature on public attitudes toward Vietnamese immigration to the UK.N = 4,376). Structural topic modeling discovered 20 latent topics, which we then used in a “deep reading” of selected tweets  migration narratives, stereotypes, and victim identities, and (2) border control. Our analysis of the news and updates cluster suggests that Twitter users amplified the spread of news as events unfolded. Alongside sharing the news, some users added their concerns and sympathies regarding the incident.This paper shows the strengths of Twitter as a platform for public opinion research. As Twitter users responded to an event in real-time, their tweets expressed immediate reactions to the tragedy, which are often implicitly and explicitly tied to immigration debates more broadly. Tweets encapsulated diverse opinions and attitudes toward complex issues such as immigration policies in only 280 characters. We show that some Twitter users, even when expressing sympathies toward victims, reproduce racist and xenophobic tropes through stereotyping and anti-immigration narratives, such as the belief that immigrants abuse the UK's social welfare system.Our research highlights the importance of a nuanced understanding of how pro- and anti-immigration attitudes manifest on social media in response to a tragedy involving immigrants. The qualitative analysis of tweets shows that Twitter users expressed both anti- and pro-immigration attitudes. Discussions shifted along with new information that emerged from further investigation. A major debate in the first few days was about the Western media's misidentification of the victims' national identities. Here, tweets critiqued the Western media as too quick to blame China for the tragedy. The discussion then shifted to expressions of sympathy to victims because they came from an impoverished region in Vietnam. Twitter users also engaged in counter-narratives that challenged the mainstream media's reporting of the event. We highlight these counter-narratives because they show “resistance and reclamation” of communities at the margins . The Vietnamese community in the UK could have tweeted in Vietnamese instead of English, and by defaulting to English, we potentially excluded their points of view. Excluding tweets in Vietnamese may have produced unknown sampling bias. Finally, although Twitter is a useful source for examining public conversations, there is evidence that data collected via the Twitter API could be biased. The API only allows access to 1% of the total tweets posted in real time. The reason for the selection of this 1% threshold is unknown to researchers outside of Twitter. This filter could create additional unknown biases.Furthermore, the relatively small number of tweets in our dataset might point to a finding itself: the lack of coverage of Vietnamese immigrants to the UK. Both the rather small amount of Twitter coverage and the fact that Twitter responses decreased relatively quickly within 2 weeks further points to the public's lack of interest in Vietnamese immigrants to the UK and their continued invisibility Barber, .Despite limitations in data collection, and modeling, our study offers rich interpretations of large-scale conversations on Twitter. Our analysis offers a unique contribution to the literature on attitudes about immigration by using social media to study public opinion about immigration issues and by providing in-depth analysis of how an unsupervised machine learning method could uncover complex discourses around immigration and border issues.https://github.com/ngathan/essex_lorry. Data is available upon request. Other inquiries can be directed to the corresponding author.The code for reproducing our results can be found at NT: study conception and design and data collection. NT and FW: analysis and interpretation of results. NT, FW, and LS: draft manuscript preparation. All authors reviewed the results and approved the final version of the 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.


In magnetic resonance imaging (MRI), this MRI is used for the diagnosis of the brain. The dynamic of these particles occurs under the action of the peristaltic waves generated on the flexible walls of the brain. Studying such fluid flow of a Fractional Second-Grade under this action is therefore useful in treating tissues of cancer. This paper deals with a theoretical investigation of the interaction of heat and mass transfer in the peristaltic flow of a magnetic field fractional second-grade fluid through a tube, under the assumption of low Reynolds number and long-wavelength. The analytical solution to a problem is obtained by using Caputo's definition. The effect of different physical parameters, the material constant, magnetic field, and fractional parameter on the temperature, concentration, axial velocity, pressure gradient, pressure rise, friction forces, and coefficient of heat and mass transfer are discussed with particular emphasis. The computed results are presented in graphical form. It is because the nature of heat and mass transfer coefficient is oscillatory which is following the physical expectation due to the oscillatory nature of the tube wall. It is perceived that with an increase in Hartmann number, the velocity decreases. A suitable comparison has been made with the prior results in the literature as a limiting case of the considered problem. Moreover, the analytical solution for concentration, temperature, tangential stress, axial velocity, and coefficient of heat transfer was deduced. Hameed et al.2 discussed the peristaltic flow of the fractional second-grade fluid confined in a cylindrical tube. They found that an increase in the constant of fractional second-grade fluid results in a decrease in velocity profile for the case of fractional second-grade fluid whereas the velocity remains unchanged for the case of second-grade fluid. Under the consideration of long-wavelength, Haroun3 is devoted to the study of peristaltic transport of a fourth-grade fluid in an inclined asymmetric channel. Heat and mass transfer analysis in the mixed convective peristaltic flow of fourth-grade fluid under viscous dissipation with Dufour and Soret effects was debated by Mustafa et al.4 where the resulting coupled nonlinear boundary value problem (BVP) was solved numerically by using Keller–box method. Krishna et al.5 explored theoretically the Hall and Ion slip impacts on an unsteady laminar MHD convective rotating flow of heat-generating or absorbing second-grade fluid over a semi-infinite vertical moving permeable surface. Rasool and Wakif6 determined the impact of Cattaneo–Christov model and convective boundary on second-grade nanofluid flow alongside a Riga pattern. The governing nonlinear problem was converted into ordinary problems via suitably adjusted transformations and the Spectral local linearization method was incorporated to find the solutions to the nonlinear problems. VeeraKrishna and Reddy7 considered the transient MHD flow of a reactive second-grade fluid through a porous medium between two infinitely long horizontal parallel plates, and the transient momentum equations were solved analytically using the Laplace transform technique. Hayat et al.8 addressed the heat and mass transfer in the peristalsis of MHD third-grade material through curved configuration. Ali et al.9 displayed the peristaltic flow of a third-grade fluid in a circular cylindrical tube when the no-slip condition at the tube wall. They observed that an increase in the slip parameter decreases the peristaltic pumping rate for a given pressure rise. Tripathi10 was investigating the transportation of a viscoelastic fluid with a fractional second-grade model by peristalsis through a cylindrical tube under the assumptions of long-wavelength and low Reynolds number. Abd-Alla et al.11 described the effect of the endoscope and heat transfer on the peristaltic flow of the Jeffrey fluid across the distance between the uniform concentric tubes utilizing the assumption of the long-wavelength and low Reynolds number to approximate the governing equations of motion. Zhao12 exhibited the convection flow, the magnetic field, and velocity slip of a peristaltic motion of a fractional fluid. Javid et al.13 used the fourth-order Runge–Kutta method to analyze the interface of the thermal boundary layer, and the porosity effects were modeled using a Navier–Stokes equation with a body force term. Wahid et al.14 talked about the boundary layer flow and heat transfer on a viscoelastic fluid over a stretched surface in a porous medium with thermal radiation and viscous dissipation. Mainardi and Spada15 offered a comprehensive overview of fractional calculus-based viscoelastic models and investigated the basic fractional models in terms of creep, relaxation, and viscosity features. With the help of Hall and induced magnetic field effects, Singh and Vishwanath16 elucidated the convective flow of a viscoelastic electrically conducting fluid within an inclined channel boundary in a porous regime. The heat transfer and second-order slip impact on the MHD flow of fractional Maxwell fluid in a porous medium were explained by Amana et al.17. Also, Tripathi18 demonstrated the use of a fractional Maxwell model to investigate the peristaltic transport of viscoelastic non-Newtonian fluids in a channel. Under the influence of a uniform transverse magnetic field, Waghmode and Suneetha19 investigated the unsteady MHD rotating free convection flow of viscoelastic fluid through a porous media with simultaneous heat and mass transfer near an infinite vertical oscillating porous plate. Narla et al.20 used the fractional calculus approach to obtain an analytical solution for the flow of a viscoelastic fluid. The influences of fractional parameter, material constant, amplitude, and curvature parameter on the pressure and friction force across one wavelength are discussed numerically with the help of graphs. Tariq and Khan 21 revealed the behavior of second-grade dusty fluid flowing through a flexible tube whose walls are induced by the peristaltic movement implementing the regular perturbation technique to get the solutions. The electro-osmotic peristaltic flow of a viscoelastic fluid through a cylindrical micro-channel was studied by Guo and Qi22 where the analytical solutions of pressure gradient, stream function, and axial velocity were explored in terms of Mittag–Leffler function by Laplace transform method. Tripathi and Bég23 applied Caputo’s definition to determine approximate analytical solutions of inclined tube peristaltic flow of a fractional second-order biofluid. Imran et al.24 discovered the effect of heat and mass transfer on particle–fluid suspension for the Rabinowitsch fluid model with the stiffness and dynamic damping effects through Darcy–Brinkman–Forchheimer porous medium. Furthermore, Bayones et al.25 displayed the magnetized dissipative Soret of steady viscous incompressible two-dimensional Maxwell fluid flow in a porous medium over a stretching sheet with chemical reaction and Joule heating. Abd-Alla et al.26 deliberated the impacts of the gravitational forces, buoyancy forces, and magnetic field on velocity profiles, temperature, and concentration of the magneto-hydrodynamic peristalsis of Jeffery nanofluid through porous media. El-Dabe et al.27 found numerical solutions for the axial velocity, temperature, and nanoparticles concentration of a non-Newtonian nanofluid flow with heat transfer through a non-uniform inclined channel. Moreover, the effects of partial slip and the magnetic field on the peristaltic flow of Walter's B fluid through a porous medium were debated by Dabe et al.28.Peristaltic flows of non-Newtonian fluids in the presence of a magnetic field have recently piqued attention in physiology, particularly in the form of a device known as Magnetic Resonance Imaging (MRI). This MRI is used to diagnose brain and vascular disorder, as well as the entire human body. In the description of viscoelastic properties, fractional calculus has had a lot of success. Vertical turbine pumps are a tried-and-true workhorse pump used in industrial applications all over the world. The oil and gas, chemical, petrochemical, desalination, power, and mining industries all benefit from this pump design. Vertical pumps are built to handle difficult apps and have a long history dating back to their invention in the Los Angeles more than a century ago. The effects of fractional Maxwell fluids on peristaltic with heat and mass transfer were investigated by Bayones et al.With these motivations in mind, the present investigation aims to discuss the influence of the magnetic field on the peristaltic pump of a fractional second-grade fluid in a porous vertical tube. To our knowledge, a magnetic field in the peristaltic pump of a fractional second-grade fluid is still unexplored. The nonlinear equations of viscoelastic fluid with fractional second-grade fluid are simplified using the assumptions of long-wavelength and low Reynolds number and then the resulting equations have been solved analytically and numerically. Comparisons of both the solutions are also made. In the end, the graphical results against different physical parameters have also been presented and discussed.10:Caputo's definition of the fractional-order derivative is defined as The constitutive equation for viscoelastic fluid with a fractional second-grade model is given by the following relation:Also, note that Z-axis is the coordinate along the axes of the tube see Fig. Let us consider a fractional second-grade fluid through a porous vertical tube. In the axisymmetric cylindrical polar coordinate system mentclass2pt{minim10:The equations governing the flow of viscoelastic fluid with a fractional second-grade model for axisymmetric flows in the fixed frame areUsing the transformations mentioned below to associate the moving Taking into account the dimensionless quantities as follow:The boundary conditions for the flow are defined as follows:The preceding equations are simplified to the following when the above-mentioned adjustments and non-dimensional variables (12) are taken into account:Furthermore, the hypothesis of the long-wavelength approach is also supposed. Now, ing Eqs. , 15), , w, w26), wThe heat transfer coefficient is indicated as follows:So, the solution of heat transfer is given by:The mass transfer coefficient is indicated as follows:So, the solution of heat transfer is given by:After getting analytical solutions of Eqs. –21 aboutIt can be observed clearly from Fig. Figure The findings are shown in Fig. w concerning the radial r for different models are plotted in Fig. The variations of the axial velocity z-axis for different values of heat source/sink parameter β Hartman number M Darcy number z-axis, while it decreases with increasing of Hartman number and wave amplitude. All these previous behaviors for different models of second-grade fluid, fractional second-grade fluid, and classical Navier–Stokes fluid, respectively.Figure The impacts of parameters Grashof number Q. Furthermore, peristaltic pumping takes place in this area otherwise Figure Figure Figure The axial velocity decreases and increases with the increase of The temperature increases and decreases with increasing source/sink parameter and wave amplitude.The concentration decreases with the increase of both Pressure rise decrease and increase with an increase in Grashof number and Darcy number.It is observed that frictional forces have an opposite behavior to that of pressure rise.The value of heat and mass transfer coefficient has an oscillatory behavior due to peristalsis.The existence of magnetic resonance imaging (MRI) and magnetic gadgets with a magnetic field allows for explaining the essential functions of living species. Inspired by these ideas, the current research project aims to investigate the influence of magnetic field, heat and mass transfer responses on the peristaltic pump of a fractional second-grade fluid.It is found that the magnetic field effect controls the velocity and temperature of the fluid. Hence magnetic field is used in cancer therapy.The results presented in this paper should prove useful for researchers in science and engineering, as well as for those working on the development of fractional second-grade fluid in a tube. Study of the phenomenon of the different physical parameters, material constant, magnetic field, and fractional parameter.The study examines the interaction of heat and mass transfer and peristaltic flow of a fractional second-grade fluid through a porous tube under low Reynolds number and long-wavelength approximation. Caputo's definition is used for differentiating the fractional derivatives. Analytical solutions are derived for temperature, concentration, axial velocity, pressure gradient, pressure rise, frictional forces, and coefficient of heat and mass transfer. The main achievement of physical parameters is illustrated. A side-by-side comparative analysis is performed to compare our findings between second-grade fluid and fractional second-grade fluid. Moreover, the fractional second-grade fluid model reduces to second-grade models for Supplementary Information.


Following publication of the original article , the autFundingThis research is supported by grants from the Roderick D. MacDonald Research Fund (Grant Number 20RDM004), the JLH Foundation (Grant Number 55354), and the NIH T32 fund (Grant Number T32CA096520).Meng Li and Shengbin Ye should be Department of Statistics, not Department of Biostatistics.The affiliation for Drs.


As a tool that can test insulators' surface morphology and properties, the performance index of atomic force microscope (AFM) probes is the most critical factor in determining the resolution of microscopy, and the performance of probes varies in various modes and application requirements. This paper reviews the latest research results in metal, carbon nanotube, and colloidal probes and reviews their related methods and techniques, analyses the advantages and disadvantages of the improved probes compared with ordinary probes by comparing the differences in spatial resolution, sensitivity, imaging, and other performance aspects, and finally provides an outlook on the future development of AFM probes. This paper promotes the development of AFM probes in the direction of new probes and further promotes the broader and deeper application of scanning probe microscope (SPM). AFM represents a well-established technique for the investigation of the nanosurface morphology. Applications range from mechanical and electrical sample characterization to measurements in liquids –4 and evAFM probe tips –10 are gCarbon nanotube probes have large aspect ratios and good wear resistance. They thus can better avoid artefacts and more accurately reflect the proper shape of the sample at steep locations, making them suitable for high-resolution test experiments. However, previous methods to fabricate carbon nanotube probes are complicated and poorly controlled. Cheng et al.  introducA colloidal probe –13 consiThis paper reviews the latest research results in the fields of metal probes, carbon nanotube probes, and colloidal probes, reviews their related methods and techniques and analyses the advantages and disadvantages of the improved probes compared with ordinary probes by comparing the differences in performance in terms of spatial resolution, sensitivity, and imaging, and finally provides an outlook on the future development of AFM probes.Metal nanoclusters contain a few to several hundred atoms that fill the gaps between nanoparticles and molecular compounds and often exhibit molecule-like electrical and optical properties because their size is close to the Fermi wavelength of electrons –17. MetaRT) and nano-tip height (z0). They are suggesting that clusters remain attached to the tip after extraction. These findings raise the question of whether the iterations of the extraction process lead to a columnar growth of the clusters, i.e., a more minor contribution of vdW interactions to the total force. To address these issues, Jiménez-Sánchez et al. subjected a blunt tip to multiple clustering extractions and characterized the extracted tip using F(z) spectra. As shown in F(z) curve and the high-resolution NC-AFM image were used as criteria to test the conditions of the constructed nano-tip. It can be seen from the Jiménez-Sánchez et al.  studied 2+ effectively quenched the fluorescence of DNA-templated silver nanoclusters (DNA-AgNCs), while the strong chelation between glyphosate and Cu2+ restored the fluorescence of the nanoclusters. The method showed good linearity in the concentration range of 1 ng/mL–50 ng/mL, and the detection limit reached 0.2 ng/mL. In addition, the method was also used for the determination of glyphosate in tap water samples with good recovery results. The assay has the advantages of high sensitivity and good selectivity.After preparation, metal nanoclusters have been widely used to detect metal ions, proteins, nucleic acids, and other substances, but few have been done to detect pesticides. Huang et al.  used met3+ after Ag+ doping. The fluorescence was restored after further adding ascorbic acid (AA) based on a redox reaction. A rapid detection method for Fe3+ and AA was established and successfully used to detect Fe3+ and AA in human serum. The DMT-AuAgNCs were successfully prepared using 4,6-diaminopyrimidine-2-thiol (DMT) as ligand, and the obtained clusters showed solid red light at 700 nm and sensitive burst selectivity for I−. The fluorescence was recovered to a certain extent by further adding Ag+. A rapid method for detecting I− and Ag+ was established and successfully applied to detect I− and Ag+ in actual samples. Gold and silver bimetallic nanoclusters' morphology, properties, and structure were analyzed by various characterization means. The target analytes with specific recognition of the materials were selected, a fluorescence analysis platform for this material was constructed, and the fluorescence response mechanism was further explored to construct a highly selective and sensitive silver-doped gold-based bimetallic nanocluster fluorescent probe.Compared to the above-proposed metal nanoclusters, gold-based bimetallic nanoclusters have the advantages of metal nanoclusters while their fluorescence quantum yields and stability are improved to a larger extent. Gold-silver bimetallic nanoclusters (AuAgNCs) exhibit enhanced luminescence efficiency and photostability due to Förster resonance energy transfer (FRET) and synergistic effects. Zhang et al.  synthesiNanowires have become a hot research topic as high aspect ratio structures –30. In aTay et al.  developeA faster, one-step technique for growing gold nanowires at the tips of commercial conductive AFM is proposed compared to the above methods. Bakhti et al.  grew golFollowing the preparation and application of monometallic nanowire probes, Fang et al.  proposedCarbon nanotubes (CNTs) are considered an ideal AFM tip material due to their small diameter, high aspect ratio, mechanical robustness, large Young's modulus, and well-defined structure –38. Due Nishijima et al.  proposedSince the above methods require scanning electron microscopy (SEM) monitoring throughout the transfer process, the process is relatively time-consuming. Hafner et al.  proposedBased on the above approach, a simple method was proposed to stabilize carbon nanotube fibers. Slattery et al.  preparedCompared to assembly methods, direct growth of carbon nanotubes by chemical vapor deposition (CVD) allows for increased bond strength between CNT tips and AFM probes. A pore growth method was used by Hafner et al. . The metSince the perpendicularity of the carbon nanotube probe is difficult to achieve in the above mentioned pore growth method, Edgeworth et al.  demonstrBased on the above method, Cheng et al.  proposedManufacturing nanoprobes that meet the requirements for cutting-edge mechanical properties, dimensions, and morphology is practically challenging. Modern technology has made it possible to produce macroscopic materials with certain functions by combining several components into composite materials. This direction has also developed concerning nanomaterials. Carbon nanotubes are compounded with related materials to produce probes, and the analytical performance of this type of probe is better than that of carbon nanotube probes alone.Nakabayashi et al.  proposedCompared with the above composite probes, the combination with metal nanowires also performs well. Clark et al.  presenteConductive atomic force microscopy (C-AFM) can be used to characterize the electrical properties of semi-conductive and conductive materials at the nanoscale. Slattery et al.  describeColloidal probes (CPs)  can simuColloidal gold particles –49 have AFM direct force measurements are mainly due to the colloidal probe technique's defined interaction geometry and versatility. Karg et al.  proposedZhang et al.  developeListeria monocytogenes (Lm) with an E. coli expression system, prepared colloidal gold by the trisodium citrate method, used colloidal gold as a tracer, optimized the preparation of colloidal gold probes, combined with purified scFv to prepare colloidal gold probes, and subjected to an immunoassay. The activity of the colloidal gold probe was determined by immunofiltration. The results showed that the colloidal gold probe was highly specific when applied to the immunodiafiltration method, and positive samples could be determined by direct color development of the colloidal gold probe. The preparation of a high-quality colloidal gold probe was a prerequisite for the detection of Lm, in which the pH value and the amount of labeled scFv were two critical influencing factors. Therefore, the study was optimized for both factors. Also, the colloidal gold probe preparation had to be followed by activity characterization to ensure the usability of the colloidal gold probe. The results showed that the detection limit of Lm was about 107 CFU/mL when the prepared colloidal gold probe was used in the immunodiafiltration method. It only took about 10 minutes to complete the detection process. The method is simple, rapid, and accurate and can be used to detect Lm in food. However, the probe lost its activity after storing at 4 °C for 28 d. This may be due to the small molecular mass, simple structure, and poor stability of the scFv relative to the intact antibody molecule. Therefore, subsequent research is needed to improve the stability and enhance the ease of use of the colloidal gold probe.Based on the above trisodium citrate reduction method, Liu et al.  preparedTo improve the stability and sensitivity of this type of probe, Miao et al.  proposedm/z 693). This indicates that the specificity of the colloidal gold nanoprobe is high enough to distinguish a target with only one base mismatch. This study developed a novel self-assembled bifunctionalized colloidal gold nanoprobe with a simple preparation process with high sensitivity and high specificity. This colloidal gold nanoprobe will have a wide range of applications in nucleic acid analysis, especially in oligonucleotide polymorphism typing for early disease diagnosis.Yang et al.  preparedTo achieve quantitative detection of pathogenic microorganisms and illegal additives in food, Huang et al.  establisColloidal probes in the types mentioned above are usually non-conductive spheres. Therefore, conductive gold spheres were introduced. Daboss et al.  proposedLarge colloidal probes are widely used to study the mechanics of samples with weaker interaction forces and local variations over a large contact range, and large CPs enable a better comparison of the results achieved in AFM compared to other probes, which can link macromechanics, micromechanics and nanomechanics well under certain conditions.Conventional AFM probes used for interaction force testing have sharp tips and have a small contact area with the surface contact. To study interaction properties on the surface of micro- and nanostructures, it is necessary to use more significant micron level colloidal probes to characterize the effect of structural features of the micro- and nanostructures on dopa adhesion. Zhang et al.  proposed−8 M BPA in water. This method of preparing colloidal fibers is simple and easy to operate and can be prepared on a large scale, providing new opportunities for many fields. In practical applications, due to the surface structure of colloidal fibers, these fibers can be used as SERS sensor detection for trace detection of BPA.The colloidal particles in the above method present a relatively stable colloidal suspension in water or ethanol solvent. The colloidal particles in the suspension can self-assemble into two- and three-dimensional colloidal crystals of various shapes under certain conditions. In the process of self-assembly, such fibrous colloidal crystals are small in size and usually micrometer in length due to the weak interaction forces between colloidal particles, and the introduction of templates requires precise operating conditions, leading to higher preparation costs, so it is necessary to find a template-free method to prepare regular large-size colloidal fibers. Xie et al.  reportedkTL of a topless cantilever beam by the thermal noise method when prepared in the chamber, after which FC is obtained on the substrate to measure the deflection sensitivity, and the position of the loading point is measured by optical microscopy when the sphere is attached. The process of calibrating the kTL into the manifold  is shown in Although colloidal probes have many excellent properties, there are some problems in calibrating such probes, especially large colloidal probes, where the intrinsic static deflection sensitivity and spring coefficient need to be verified. Chighizola et al.  proposedIn this method, Chighizola et al. verified that adding a large colloid at the end of a topless cantilever beam increased the quality factor and significantly reduced the resonant frequency. This type of probe can better simulate a single-mode spring-mass system. Although some adjustments are made to its cantilever dynamics, the calibration accuracy of CPs does not fluctuate when the additional mass of the ball is the same as the cantilever. In the type of large CPs, applying the homogeneity theorem for topless cantilever beams to Probe technology has been applied in physics, chemistry, biology, and nanotechnology in the past few years, and outstanding results have been achieved. Metal, carbon nanotube, and colloidal probes have been widely used. Metal plating probes are generally suitable for high-resolution or high conductivity test experiments. Carbon nanotube probes have a large aspect ratio and good wear resistance, thus can better avoid artefacts and more accurately reflect the actual shape of the vertical position of the sample, and are suitable for high-resolution test experiments. Colloidal probes are mainly used to study colloidal interactions. They can simulate the determination of the forces between various types of colloidal particles such as silica-alumina cellulose, providing a good detection tool for a better understanding of interactions in various fields. This paper reviews the latest research results on metal, carbon nanotube, and colloidal probes. The differences in spatial resolution, sensitivity, imaging, and other performance aspects are compared. The advantages and disadvantages of the improved probes compared with standard probes are analyzed in The development of AFM technology strongly depends on micro-cantilever probe preparation technology. Hence, developing probes with stable performance, high resolution, long life, and broader applications is a hot spot and complex area of current research in AFM technology. Therefore, in the field of AFM probes in the future, ultra-fine ultra-tip and ultra-long-life probes may be developed to improve the resolution and lifetime of electrical and magnetic energy probes. The nanosizing of probes, especially the modification of carbon nanotubes and functional nanomaterials, will significantly improve the performance of probes and further promote the broader and deeper application of SPM. It will significantly impact society in life science, drug discovery, and medical diagnosis.


This paper analysed the healthiness of 823 culinary recipes retrieved from 755 videos shared during a six-month period on ten popular Brazilian YouTube® cooking channels. Recipes were categorized by type of preparation. To assess recipes’ healthiness, ingredients were classified according to the extension and purpose of industrial processing, in order to identify the use of ultra-processed foods. Additionally, a validated framework developed from criteria established in both editions of the Dietary Guidelines for the Brazilian Population was employed. Recipes for cakes and baked goods, puddings, snacks and homemade fast foods, which were among the most frequently posted, contained the lowest proportion of unprocessed/minimally processed ingredients and the highest proportion of ultra-processed ingredients. Recipes containing whole cereals, fruits, legumes, nuts, and seeds were scarce. Results indicate that users should be critical about the quality of recipes shared on YouTube® videos, also indicating a need for strategies aimed at informing individuals on how to choose healthier recipes or adapt them to become healthier.Social media platforms are readily accessible sources of information about cooking, an activity deemed crucial for the improvement of a population’s diet. Previous research focused on the healthiness of the content shared on websites and blogs, but not on social media such as YouTube Public health initiatives from many countries encourage home cooking as a health-promoting strategy . This isAside from stressing the importance of home cooking, the Dietary Guidelines for the Brazilian Population published in 2015 focused on categorizing foods according to the extension and purpose of industrial processing . IndividHigh consumption of UP foods has been associated with chronic non-communicable diseases and all-cause mortality ,6,7. ConInformal cooking education happens through culinary socialization over the course of a person’s life, a process in which individuals acquire patterns of practices and perceptions related to cooking, from socializing agents . The fir®, Instagram®, Pinterest® and YouTube® to share and search for recipes, and to find meal suggestions and inspiration [Brazilians spend an average of 3.5 h daily on the Internet , mainly piration ,18,19,20® was created in 2005 and works as a video sharing platform, which is accessible via personal computers or smartphones through an Internet browser or application [YouTubelication . On the lication ,23,24.® is one of many people’s favourite ways to learn how to cook [® is the most popular social media platform among individuals aged between 16 and 64 years [Previous research mentions that YouTube to cook . Underst to cook . Video r to cook . In BrazAccessing the Internet to search for recipes, learn how to cook, and develop cooking skills is recommended by the Dietary Guidelines for the Brazilian Population , but the® as a data source [Previous studies assessed the healthiness of Internet recipes on websites and blogs , and concluded that users tend to interact more often with the least healthy recipes . Authorsa source . The papa source . No rese® cooking channels from Brazil, using both national dietary guidelines as references. We adopted complementary approaches to assess recipes’ healthiness, the first being the analysis of recipes’ ingredients according to the extension and purpose of industrial processing, an important and widely used approach to categorize foods. Subsequently, a specially designed qualitative framework was used to characterize recipes according to cooking method, and by the presence of healthy or unhealthy ingredients. We believe this study has the potential to inform the design of public health initiatives that guide individuals and inform dietitians on how to select and critically evaluate sources of cooking information, and improve the quality of homecooked meals.To address this gap, this descriptive and exploratory study analyses the healthiness of culinary recipes shared on popular YouTubeConsidering the scarcity of literature on the research topic, a pilot study was carried out to inform the data collection protocol, which included aspects of various channels’ eligibility criteria and database layout, introduced different video characterization variables, validated recipes’ assessment method, and determined the data collection period, taking into account the temporal feasibility of the study and the amount of content to be analysed .https://www.channelcrawler.com/, accessed on 10 February 2020), which classifies channels according to criteria established by the researcher . During the pilot study, it was observed that cooking channels belonged to the ‘How to and Style’ category on YouTube®, thus, all channels of the platform within that category were accessed in decreasing order of subscribers to identify which ones best fit the eligibility criteria. The ten biggest channels which (1) presented audio-visual content in Portuguese and were Brazilian based; (2) were a cooking channel; (3) posted culinary videos at least once a week; and (4) were not an advertising channel or reproduced television cooking programs were selected (Cooking channels were purposely selected by taking into consideration that popularity (number of subscribers) can promote a greater reach and be a proxy for users’ preference. Channels were selected according to the number of subscribers in February 2020 using The YouTube Channel Crawler page , two by men, and one by a couple; none of them were popularly known chefs or food celebrities. Subscribers ranged from 514 thousand to 4.25 million; channels’ time of existence ranged from 4 to 9 years, and posting frequency varied from 2 to 7 videos per week.The included channels were mostly presented by women  posted during a six-month period (from February to August 2020) on ten different cooking channels was selected. Considering that this is a recent field of study and there is no specific recommendation in the literature for how long data collection on YouTuben = 35), (2) presented a festive recipe , (3) were sponsored by the food industry (n = 9), (4) were a repost (n = 7), (5) presented recipes linked to the COVID-19 pandemic .All videos with recipes posted within the period were watched in full (first author) to determine if they contained all the ingredients needed, as well as the preparation method. A total of 106 videos were excluded from analysis because they did not meet the eligibility criteria: (1) were recorded live transmissions (Reasons for not including recorded live transmissions were: (1) during the pilot study, we observed that those kinds of videos were usually presented as ‘extra’ content and were produced by only four of the ten channels. They were not included for the sake of standardization (type and number of videos per channel). (2) ‘Live’ transmissions lasted more than one hour each, as the recipe-related content was diluted among various other content during the video. This affected both the practical relevance of the recipe and the temporal feasibility of the research.® was created to include the following information for each video: title, access link, ID provided by YouTube®, video description, date of posting, date of access, duration in seconds, number of likes, dislikes, and views. To obtain the number of comments posted by users in each video, a command line application was developed in Python 3.0 (third author). Using the public and free Google Data application programming interface (API) service® as the data source, the application generated automated reports from the video ID and the period determined by the researcher .Weekly, from February to August 2020, each selected channel was accessed via computer and all videos posted during the previous week were registered. A database in Microsoft Excel 2016Next, the first author watched each video to register the ingredients and the cooking method in the database. If further details about ingredients were needed, the researcher consulted the recipe’s ingredient list provided in the video description, or, in case of industrialized products, the packaging, when information was clearly visible on screen. Steps or ingredients mentioned by the youtuber as ‘optional’ and not shown in the video were not assessed.Variables assessed to characterize the videos were duration in minutes, day of the week of posting, and interaction measures including popularity as daily views in the first week, approval as daily likes and dislikes in the first week, direct interaction of users with content through daily comments in the first week, and total comments in the first week and in the first month after the video was posted.® by determining the degree of similarity of the words and phrases used and the characteristics of the recipes, starting at coding recipes’ names in videos’ titles (first author). After coding, data was categorized until strong or terminal categories appeared [To classify the recipe into a category  a content analysis was carried out based on the video’s title, description, and list of ingredients used. This analysis was manually organized in Microsoft Excel 2016appeared .Recipes had their ingredients classified according to the extension and purpose of industrial processing, as unprocessed/minimally processed (U/MP), processed culinary ingredient (PCI), processed (P), or ultra-processed (UP) (first author) ,4,33. InSubsequently, the Qualitative Framework for the Assessment of Culinary Recipes’ Healthiness  was applTo ensure data quality control, the second author independently analysed 10% of the recipes from the dataset. Weighted kappa of agreement between raters for the assessment of ingredients’ extension and purpose of industrial processing was 0.96, and ranged between 0.90 and 1.00 (kappa and weighted kappa) for the application of the Qualitative Framework for the Assessment of Culinary Recipes’ Healthiness, indicating almost perfect agreement in both analyses . ContentQualitative dichotomous and polytomous variables are presented in absolute and relative frequencies. Quantitative variables are presented as median and interquartile range (IQR), considering the non-normality in data distribution when assessed by Shapiro–Wilk test, histogram, kurtosis value, and mean/median proximity.n = 141) vs. during social isolation (n = 614) . Mann–Whitney and Kruskall–Wallis tests were used for quantitative variables. For qualitative variables, Pearson’s chi-square test was employed. Stata 13.0®  was used for analysis and a post-hoc power analysis was applied on G*power 3.1.9.2 whenever necessary, considering a two-tailed test. An alpha of 0.05 was established as the significance level for all analyses.Variables of videos’ characteristics and recipes’ healthiness among the categories of recipes were compared. Also, as data collection took place mostly during a social distancing period due to the COVID-19 pandemic, when searches for recipes online increased , we alson = 755). The number of daily likes in the first week was superior to daily dislikes. The option of liking or disliking a video was not enabled by the youtubers for all videos (only n = 611), therefore, even if users wanted to give a particular video a thumbs-up or down, they could not. Direct interaction through comments was concentrated in the first week after the videos were posted, as the median of total comments in the first month was close to the median in the first week. Sunday was the day of the week with the lowest number of videos posted, nevertheless, the distribution of videos was similar among the other days .The only observed difference between videos collected in the period preceding vs. during social isolation was in the total of comments in the first week, which was higher during the social isolation period  than before the pandemic  . The frequency of categories observed in the period preceding vs. during social isolation was statistically the same .More than two thirds of all recipes (68.1%) comprised preparations from only four categories, namely: meat or egg main dishes; cakes and baked goods; snacks and homemade fast foods; and puddings . The sixn = 4242) and PCI . Ingredients classified as P  and UP  were less frequent. The categories of recipes differed in terms of the ingredients’ distinct extension and purpose of industrial processing . As 2 = 0.68; p = 0.877).Of the total 7814 ingredients analysed, the majority were U/MP . Mentions of tomato sauce with herbs (bottled or freshly made) were more frequent than exclusive mentions of white sauce with mayonnaise or cheese . Exclusive use of industrialized seasonings  and of frying as a cooking method  was also not frequently mentioned. On the other hand, the presence of whole cereals, breads and/or pasta, either exclusively or mixed with refined cereals was low in the recipes , as well as were the presence of fruits , legumes , and nuts and seeds . The categories that presented the most evenly distributed positive and negative criteria were types of meats, presence of foods with high sugar concentration, and presence of vegetables. All results from the framework analysis were statistically the same regarding the period of data collection .Application of the Qualitative Framework for the Assessment of Culinary Recipes’ Healthiness  identifi® cooking channels from Brazil using the Dietary Guidelines for the Brazilian Populationas references. Recipes posted during a six-month period were retrieved and categorized into sixteen different groups. The most frequently posted recipes were of meat/egg-based main dishes; cakes/baked goods; snacks/homemade fast foods; and puddings. This means that recipes for salads and side dishes, which usually contain vegetables, fruits, and legumes, were shared less often than recipes with animal sources of protein, all-purpose flour, fats, and sugar as the main ingredients. This result is not favourable from a health standpoint, as individuals are possibly being led to prepare fewer recipes with fruits, vegetables, and legumes, which are linked to a lower risk of chronic non-communicable diseases, and are largely present in most healthy eating patterns [® is a multi-content platform not specifically focused on recipes, our recipe categories were qualitatively and inductively generated from recipes’ titles, descriptions, and ingredients. Additionally, several studies only assessed specific categories of recipes [This study analysed the healthiness of recipes shared on popular YouTubepatterns ,38. Intepatterns ,39 and wpatterns , recipes recipes ,29,30,39® [®, every culinary preparation had statistically equal measures of interaction , possibly due to differences between the profiles of users from recipe websites [®, as a video platform, enables a kind of interaction that gives users a feeling of being connected not only to a video, but to a person who shares their beliefs and interests. This feature can promote a certain measure of social bonding in which people feel connected with one another and start following the channel for further communication. For user-created content such as the videos analysed, a sense of community is fundamental; so it is possible that subscribers give the same attention to a recipe, regardless of whether it is a salad or a cake, in order to provide support through constancy of viewership and interaction [Another possible explanation for the low prevalence of fruit- and vegetable-based recipes in our sample may be that content producers expect users to interact with the postings through comments and shares, as interaction is fundamental for a channel’s engagement and sustainability . It has ®  that intwebsites  and evenwebsites . YouTubeeraction . The reaRecipes’ ingredients were mainly U/MP foods and PCI, so one can argue that from a wide perspective, the recipes could lead individuals to cook recipes that are aligned with the recommendations of the Dietary Guidelines for the Brazilian Population . NeverthThe presence of UP ingredients in the recipes may be explained by their convenience appeal ,33. It i® reach a wide audience, this finding reinforces the need to not only encourage people to look for recipes online [While the majority of recipes were healthy with respect to avoiding the use of margarine, avoiding frying, and opting for sauces with lower fat content, other aspects such as incorporating whole cereals, fruits, legumes, nuts, and seeds in preparation were not frequently present. Considering that social media platforms such as YouTubes online , but alss online , while rs online . We are s online . MembersThe adoption of a conservative criterion for classifying ingredients by the extension and purpose of industrial processing may have led to an underestimation of the number of UP ingredients. Nevertheless, this approach mirrors how information reaches users—they also do not necessarily have access to information on labels when watching videos.Data collection took place during months of social isolation due to the COVID-19 pandemic, when searches for recipes online increased . This wa® channels’ popularity oscillates constantly. To handle this, we repeated the channel selection step at the end of data collection, and verified that they remained as the ten most popular in the period, despite some outperforming others in the number of subscribers.YouTube®’s own algorithms, a view might not indicate a user who has watched the content in its entirety. Content approval (likes and dislikes) also does not indicate whether or not an individual fully watched the content before giving a positive or negative rating. Although views made by computer programs rather than by humans are not counted [As the number of views is validated by YouTube counted , those i counted .In the context of television cooking shows, some researchers argue that the consumption of this content is unlikely to impact habitual dietary intake, because entertainment and leisure are the main reasons people watch those programs ,49. NotwWe understand that it is not possible within the confines of the present study to account for variations in the reproduction of recipes at home, such as instances when people do not follow all the steps, or when ingredients are exchanged, which may result in a different assessment of their healthiness.As positive points, we highlight the investigation of culinary recipes posted in the most used social media in Brazil, by adults ; the rig®, users and subscribers to popular cooking channels should be aware that most recipes are based on ingredients such as meats, eggs, all-purpose flour, fats and sugar, and only a few have whole cereals, fruits, legumes, nuts, and seeds. Recipes for puddings, cakes and baked goods, snacks and homemade fast foods, sauces, sweet and savoury spreads, and pâtés had, in addition to low numbers of U/MP food ingredients, higher numbers of UP foods as ingredients. Our findings can inform health professionals and policymakers on how to promote healthier culinary recipes, how to interact with content creators, and how to advise individuals about the quality of the recipes shared on YouTube® videos, and hence, can help them choose healthier recipes or teach them how to modify the recipes into healthier versions. Future research exploring how users from different populational groups interact with culinary content on distinct social media platforms will be relevant for advancing this field of study.This study provides a comprehensive overview of the healthiness of culinary recipes shared on a social media platform, one of the favoured avenues for the search of cooking-related content. On a professional practice and health promotion note, although it is praiseworthy that people are cooking and sharing their knowledge on platforms such as YouTube


Cellular & Molecular Immunology 10.1038/s41423-022-00928-4, published online 27 October 2022Correction to: In this article the statement in the Acknowledgements section was incorrectly given as “MRB and IIS are supported by the NIH R01GM080139 grant (to IIS), the Welch Foundation Research Grant AU-2014-20190331 (to IIS), and the American Heart Association grant 18CDA34110086 (to MRB)” and should have been, “MRB and IIS are supported by the NIH R01GM072804 grant (to IIS), the Welch Foundation Research Grant AU-2014-20190331 (to IIS), and the American Heart Association grant 18CDA34110086 (to MRB).”The original article has been corrected.


Dear Editor,p = .026; Fig. Although previous studies reported associations between gut bacterial dysbiosis and asthma,Malassezia and Aspergillus. Linear discriminant analysis effect size was used to investigate mycobiome biomarkers (discriminately enriched fungi) at various taxonomic ranks. This revealed that one class, three families, and three genera, including Fusarium, were discriminately enriched in HC. One phylum, one class, three families, and eight genera  were enriched in the NT group, the polysaccharides of Dictyophora was demonstrated to have several physicochemical and biological properties.Russula is a valuable, medicinal fungus with anti‐inflammatory bioactivities.Ganoderma, Russula, and Senacina and negatively with the abundance of Saitozyma and Fusarium. Sputum neutrophils (%) were positively associated with the abundance of Mortierella, Didymella, Saitozyma, Penicillium, and Pyrenochaetopsis , evenness (Pielou_e), and diversity (Shannon), and mild‐to‐moderate patients showed significantly lower fungal diversity (Shannon) Fig. . PCoA inp < .05, nine (ESI+) and three (ESI–) discriminate metabolites were found between NT and HC groups . Therefore, these exploratory findings require validation through large‐scale studies.In conclusion, asthma patients harboured altered gut fungal compositions, metabolic profiles, and associated metabolic pathways. ICS altered the mycobiome community and fungal/bacterial–metabolite connections. Our findings further our understanding of the gut mycobiome, metabolome, and fungal/bacterial–metabolite associations that may be critical in asthma pathogenesis and may therefore be therapeutic targets.The authors declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported.the National Natural Science Foundation of China, Grant Numbers: 82170023 and 81970020; Shanghai Sailing Program, Grant Number: 20YF1428300; Shanghai Municipal Health Commission, Grant Number: 2019SY006; Shanghai Key Laboratory of Emergency Prevention, Diagnosis and Treatment of Respiratory Infectious Diseases, Grant Number: 20dz2261100 ; Shanghai Municipal Key Clinical Specialty, Grant Number: shslczdzk02202; Cultivation Project of Shanghai Major Infectious Disease Research Base, Grant Number: 20dz2210500; Shanghai Key Discipline for Respiratory Diseases, Grant Number: 2017ZZ02014; Shanghai Shenkang Hospital Development Center Clinical Science and Technology Innovation Project, Grant Number: SHDC12018102Supplementary MaterialsClick here for additional data file.Supplementary Figures and TablesClick here for additional data file.


Baby boxes provide goods to new parents and a space for infant sleeping. They were first introduced in Finland, and it has been argued that the policy helped reduce infant mortality. We evaluated the impact of the Finnish Maternity Grant (which includes the Finnish Baby Box) on infant mortality rates (IMRs) at the points of introduction (disadvantaged mothers only) in 1938 and universalisation in 1949.Maternity Grant introduction and universalisation were evaluated as distinct natural experimental events, using interrupted time series analysis. The outcome was IMR per 1000 live births. We analysed national data on all infants born in Finland between 1922 and 1975, estimating step and trend changes in the outcome following the point of intervention. Sensitivity analyses included truncating the pre-intervention period and a double break point model, incorporating terms for both introduction and universalisation.Maternity grant introduction in 1938 was associated with a step-change increase  in Finnish IMRs. Maternity grant universalisation in 1949 was associated with a step-change decrease  in Finnish IMRs. Sensitivity analyses produced corresponding associations.While we observed changes in IMRs associated with Maternity Grant introduction and universalisation, these changes cannot be disentangled from the impact of the Second World War or other relevant policy developments on infant mortality. Consequently, the relationship between the Finnish Baby Box or comparable contemporary interventions and infant mortality remains unclear. Interventions modelled on the Finnish Baby Box have seen increasing international uptake, often alongside claims relating to infant mortality and sudden infant death syndrome.Despite these claims, our study could not reliably estimate the impact of the Finnish Maternity Grant (which includes the Finnish Baby Box) on infant mortality.Our study underscores that the causal relationship between baby boxes and infant mortality remains unclear. However, future research should consider if other health benefits might still be possible.The Nordic nations were forerunners in Europe’s 20th century infant mortality decline, despite having comparatively low levels of economic prosperity.Speculation has recently surrounded the causal role of the Finnish Baby Box in the decline of Finnish IMRs.The Maternity Grant was first introduced in Finland in 1938 and later universalised in 1949. Concerns around low birth rates and infant mortality were prominent in Nordic countries at this time.Our evaluation of the Maternity Grant aimed to estimate the impact of both introduction and universalisation in 1938 and 1949, respectively, on Finnish IMRs. We hypothesised that, if the Maternity Grant did indeed have a beneficial impact on IMRs, we were most likely to observe this effect following 1938. This was argued from the fact that it impacted a larger proportion of the population than in 1949 and that it targeted disadvantaged mothers, who may be expected to benefit more from receiving the Maternity Grant.The introduction and universalisation of the Maternity Grant were evaluated as distinct natural experimental events, using interrupted time series analysis (ITS). ITS is a quasi-experimental method, widely used in the evaluation of healthcare and population-level health interventions.10.1136/jech-2022-219488.supp1Supplementary datahttps://www.mortality.org/).All data used in this evaluation were sourced from the Human Mortality Database (HMD)  and a postintroduction period from 1938 to 1953 (16 years) were chosen; for Maternity Grant universalisation in 1949, a preuniversalisation period from 1922 to 1948 (27 years) and a postuniversalisation period from 1949 to 1975 (27 years) were chosen. In each case we sought maximise the preintervention data used, while permitting the inclusion of the UK as a control population in the SC analyses (UK data were only available from 1922 onwards).We used a segmented linear regression to measure the preintervention trend, the immediate step change following the intervention and the change in the postintervention trend compared with the preintervention trend.https://github.com/MacCaibe/Maternity-Grant-Evaluation.All analyses were conducted using R V.4.0.3. The data and code for this analysis are publicly available at: et al.For both Maternity Grant introduction in 1938 and universalisation in 1949, we truncated the preintervention period to 10 years. This sought to understand whether historic trends were consistent with more recent trends. For the 1949 intervention point specifically, a quadratic term was added to the model retrospectively to capture the curvilinear form of the postintervention outcome trend. Finally, a segmented regression with a double break point incorporating both intervention points was used as an alternative to modelling each intervention point in isolation. As an alternative to ITS analysis, a preliminary SC analysis was performed for both Maternity Grant introduction and universalisation following Abadie and GardeazabalThe Finnish IMR was high throughout the first half of the 20th century . In 1937The main ITS model for Maternity Grant introduction estimated an immediate step increase of 14.59 (95% CI 4.30 to 24.89) deaths per 1000 live births, alongside a marginal and imprecise trend decrease . Step anThe main ITS model for Maternity Grant universalisation in 1949 was associated with an immediate step decrease of −14.35 (95% CI −20.94 to −7.76) deaths per 1000 live births, alongside a marginal trend increase . In 1950We evaluated each intervention point as a natural experiment using ITS analysis. Infant mortality was seen to rise following Maternity Grant introduction in 1938. Instead of the Maternity Grant, this may resemble the impact of the Continuation War from 1941 to 1944 and the Lapland War from 1944 to 1945 which likely increased IMRs . Maternity Grant universalisation in 1949 was associated with a decrease in IMRs. However, the influence of other events such as the Child Allowance System—which provided a monthly tax-free cash transfer to mothers from 1948 onwards—and the roll-out of general antenatal care on infant mortality were not accounted for. The latter came into effect in 1945 and likely contributed to falling infant mortality; 86.4% of mothers attended antenatal care in 1945 compared with just 31.3% in 1944.We provide the first empirical evaluation of the relationship between the Finnish Baby Box and infant mortality, using high-quality data and robust methods of causal inference. This can be viewed as an example of public health science engaging with a wider policy debate.Owing to a lack of individual-level data, however, we were unable to isolate the effect of the Finnish Baby Box specifically (as opposed to the wider Maternity Grant). It also meant that, for the evaluation of Maternity Grant introduction, a sizeable proportion of the study population classified as exposed were actually unexposed until universalisation 1949. Conversely, for the evaluation of Maternity Grant universalisation, a sizeable proportion classified as unexposed were actually exposed since introduction 1938. Another major obstacle to causal inference in this study was variability in the outcome trend and the presence of other events relevant to the outcome. For example, it was not possible to account for either the events of the Second World War or other policy developments . Finally, SC models were not interpretable, suggesting a suitable control population (synthetic or otherwise) was not available for this evaluation; this is highlighted by the dissimilarity of the Finnish IMR trend compared with that of the control regions included in the SC models see . These lIt cannot be assumed that policies modelled on the Finnish Maternity Grant or Finnish Baby Box reduce infant mortality, as there remains no evidence clearly demonstrating this. The transferability of the Finnish Baby Box to modern contexts should be carefully considered. For example, IMRs were higher in Finland at the time of Maternity Grant introduction than in most low-income and middle-income countries at present.The introduction and universalisation of the Maternity Grant were associated with changes in IMRs. However, these associations cannot be interpreted as causal as we did not account for of the impact of the Second World War or other relevant policy developments  on infant mortality. As such, the relationship between the Finnish Baby Box or similar interventions and infant mortality remains unclear.


Clearance of ocular fluid and metabolic waste is a critical function of the eye in health and disease. The eye has distinct fluid outflow pathways in both the anterior and posterior segments. Although the anterior outflow pathway is well characterized, little is known about posterior outflow routes. Recent studies suggest that lymphatic and glymphatic systems play an important role in the clearance of fluid and waste products from the posterior segment of the eye. The lymphatic system is a vascular network that runs parallel to the blood circulatory system. It plays an essential role in maintenance of fluid homeostasis and immune surveillance in the body. Recent studies have reported lymphatics in the cornea , ciliary body, choroid, and optic nerve meninges. The evidence of lymphatics in optic nerve meninges is, however, limited. An alternative lymphatic system termed the glymphatic system was recently discovered in the rodent eye and brain. This system is a glial cell-based perivascular network responsible for the clearance of interstitial fluid and metabolic waste. In this review, we will discuss our current knowledge of ocular lymphatic and glymphatic systems and their role in retinal degenerative diseases. The eye is a sensory organ responsible for the initial stages of visual transduction. It is filled with a transparent water-like fluid called aqueous humour within its anterior fluid compartments, which plays an essential role in the normal functioning of the eye. Dysfunction of the aqueous drainage system has been associated with a number of ocular disorders. It is generally considered that aqueous humour leaves the eye via two routes in the anterior eye, called the trabecular and uveoscleral pathways; these routes pass close to the lens and iris ,2. HowevThe lymphatic and glymphatic systems have been suggested to play an important role in clearing interstitial fluid and metabolic waste from the retina, which is located in the posterior segment of the eye, as well as the optic nerve, which relays action potentials to the brain for processing. Despite their potentially important roles in fluid regulation, the presence and function of lymphatic and glymphatic systems in the eye are only just beginning to be unravelled. Improving our knowledge of ocular fluid drainage pathways is essential for better understanding of eye function in health and disease.In this review, we will first provide an overview of ocular fluid aqueous humour, including its drainage routes and retinal fluid regulation pathways. We will then review current understanding of lymphatic and glymphatic systems and evidence for their presence in the eye. Finally, we will discuss the role of lymphatic and glymphatic systems in the pathogenesis of ocular disorders with particular focus on retinal degenerative diseases.Aqueous humour (AH) is a clear, slightly alkaline fluid contained in the anterior and posterior chambers of the eye, and is comprised of water, ions, carbohydrates, and proteins . AH is pAH is continuously formed by the ciliary epithelium of the ciliary processes projecting from the pars plicata (anterior portion of the ciliary body) , at a tySecretion of aqueous humour involves active transport of ions and other molecules through the non-pigmented epithelium of the ciliary epithelium into the posterior chamber, which in turn creates an osmotic gradient across non-pigmented epithelium (NPE) cells. This osmotic gradient then facilitates the passive water movement into the posterior chamber via aquaporin (AQP) water channels expressed by NPE cells: AQP1 and AQP4 ,12. AfteAH exits the eye through two pathways, called conventional and unconventional pathways . In the In the unconventional pathway, aqueous humour diffuses through the ciliary muscle, supraciliary (space between ciliary body and sclera), and suprachoroidal (space between choroid and sclera) spaces . From thAlthough the bulk of AH exits the eye via trabecular and uveal routes, some portion of AH ~20%) flows posteriorly and enters the vitreous chamber in response to intraocular pressure ,20,21. A% flows pThe regulation of extracellular fluid is critical to retinal function in health and disease. Failure of retinal water regulation can cause retinal disorders such as retinal oedema . In normMüller glia remove water from the inner retina by facilitating water transport to the vitreous humour and retinal capillaries ,27,28. T+ currents that flow through these cells [+ and water [+ through their K+ channels from the retina to the vitreous body and blood; this process is known as K+ spatial buffering or K+ siphoning [+ channels in Müller cells responsible for K+ siphoning is the inwardly rectifying K+ channel 4.1 (Kir4.1) [+ from perisynaptic spaces to blood vessels [Considerable evidence indicates that the water-transporting function of Müller cells is coupled to Kse cells ,33,34,35nd water ,36. Mülliphoning ,37,38. T(Kir4.1) . The Kir(Kir4.1) ,33. This vessels ,40,41.Water from the subretinal space is cleared by transcellular ion and water fluxes through RPE cells . In the The lymphatic system represents a second circulatory system, parallel to the blood vascular system. In humans, the lymphatic system is distributed throughout the body, with the exception of the central nervous system (CNS) . The lymThe network of lymphatic vasculature is composed primarily of blind-ended and thin-walled capillaries. These lymphatic capillaries are lined by a single layer of overlapping endothelial cells that have a distinctive oak leaf-like shape . AlthougIn contrast to lymphatic capillaries, collecting lymphatic vessels are made up of spindle-shaped endothelial cells. These vessels have a complete basement membrane and are encircled by one or two layers of smooth muscle cells ,53,54. MLymphatic vessels maintain tissue fluid homeostasis by clearing interstitial fluid. At increased interstitial fluid pressure, lymphatic capillaries collect excess extracellular fluid and pass this fluid, now called lymph, to collecting lymphatic vessels. From these vessels, lymph then flows into and out of lymph nodes via afferent and efferent lymphatic vessels, respectively. Finally, lymph drains into blood circulation through the lymphaticovenous junction between the thoracic or lymphatic duct and the subclavian veins ,49,56,57The lymphatic system performs several important functions, including tissue fluid balance, dietary fat absorption, and immune surveillance . LymphatDespite the availability of several antigenic markers, unequivocal identification of lymphatic vasculature remains challenging, particularly in the eye. Lymphatic capillaries are often irregular and collapsed, thus making it difficult to visualize them histologically . MoreoveThe eye was long considered to lack lymphatics except for the conjunctiva . PreviouIn the posterior part of the eye, lymphatic capillaries have been identified in the developing and adult human choroid . HoweverIn summary, it appears that the presence of ocular lymphatics, particularly in the uvea, remains highly debated. Based on the critical literature review, it is clear that most of the studies on ocular lymphatics did not use (1) both the structural and functional experimental approaches or (2) both the multiplex immunolabeling and ultrastructural (including immunogold labelling) studies. Therefore, future studies could be extended to multi-modal experimental approaches to confirm the presence or absence of ocular lymphatics.Although CNS has long been thought to lack a classical lymphatic vasculature, recent studies have discovered a quasi-lymphatic system (termed the glymphatic system) in the brain parenchyma , as wellCerebrospinal fluid (CSF) is a clear, proteinaceous fluid that occupies the ventricles of the brain and the sub-arachnoid space surrounding the brain, spinal cord, and optic nerve. The CSF volume in adult human is estimated to be 150 mL, with 125 mL in the sub-arachnoid space and 25 mL in the ventricles . The CSFPial arteries and veins run through the sub-arachnoid space and are surrounded by CSF. Among these blood vessels, only pial arteries are enveloped by pia mater . These aThe perivascular space is a fluid-filled compartment that surrounds the blood vessels (arteries and veins), penetrating into the brain parenchyma . This spAQP4 knockout mice. Using in vivo two-photon and ex vivo fluorescence imaging, they found that CSF transport into and through the brain interstitium is significantly reduced in AQP4−/− mice compared to wild-type control mice. Moreover, they observed that interstitial solute clearance from the brain was reduced by ~70% in AQP4−/− mice [AQP4 knockout mice [AQP4 is the predominant water channel protein in the brain, where it is expressed by astrocytes and ependymal cells . Astrogl−/− mice . The impout mice ,125,126.out mice ,126,127.As part of the CNS, the eye—particularly the retina and optic nerve—shares embryological, anatomical, and physiological similarities to the brain ,129. In Growing evidence suggests that ocular lymphatic and glymphatic systems are involved in the pathogenesis of a number of disorders associated with ocular fluid homeostasis and waste clearance. The following section reviews current research investigating the role of lymphatic/glymphatic drainage in retinal diseases.Glaucoma is a multifactorial optic neuropathy characterized by the slow progressive degeneration of retinal ganglion cells (RGCs) and their axons within the optic nerve. This degeneration results in structural changes in the optic nerve head and subsequent visual field defects, leading to blindness ,139. It The lymphatic system has been suggested to play a role in glaucoma pathogenesis due to its involvement in aqueous humour drainage. One recent study showed that angiopoietin (ANGPT) growth factors are critical for the development of lymphatic vessels in the corneal limbus, and the loss of ANGPT1 and ANGPT2 from lymphatics is associated with glaucoma and ocular hypertension in mice . ImpaireThe glymphatic system has also been implicated in the pathophysiology of glaucoma. Following intravitreal injection of human amyloid beta in glaucomatous rodent eyes, glymphatic clearance of amyloid beta from the retina decreased significantly . MoreoveAge-related macular degeneration (AMD) is a retinal disease associated with aging that gradually affects the macula—a specialised region in the retina responsible for sharp central vision needed for simple everyday activities . The patThe ocular glymphatic system has been hypothesized to be involved in AMD pathogenesis. This hypothesis is based on the facts that (1) the glymphatic pathway clears metabolic waste, including amyloid beta peptide, from the CNS and AMD is associated with amyloid beta deposition ,148, andRecent studies suggest that an impaired ocular lymphatic system due to ageing might be associated with ocular inflammatory diseases such as uveitis ,150. HowEmerging evidence indicates that physiological, pharmacological, and lifestyle interventions may help improve CNS glymphatic function under physiological and pathological conditions. Over the past few years, natural sleep has been implicated in glymphatic function under physiological conditions. In a recent study on a rodent model, Xie et al. found that natural sleep significantly increased CSF–ISF exchange in mouse brain parenchyma with a consequent increase in the glymphatic clearance of amyloid beta . Under pGiven that CNS glymphatic function is facilitated by astrocytic AQP4 water channels, a number of AQP4 inhibitory and facilitatory compounds have been investigated for their modulatory effects on glymphatic function. In a rat stroke model, AER-271  has been found to reduce cerebral oedema, suggesting that this compound might be used to improve the glymphatic function under neurological conditions. This suggestion was based on the fact that glymphatic system has been reported to be associated with cerebral oedema formation following ischemic stroke . Other AA recent study on a rodent model of Alzheimer’s disease was carried out to investigate the relationship between exercise (a lifestyle factor) and glymphatic function ,158. TheIn contrast to the CNS glymphatic system, limited studies have been carried out investigating the modulation of similar systems in the eye. In a recent study, Wang et al. demonstrated that light-induced pupil constriction increased the glymphatic clearance of intravitreally injected human amyloid beta from mouse retina . MoreoveSeveral recent studies reported the potential of lymphatic modulation in the treatment of retinal diseases. Regulation of lymphatic trafficking has been reported to promote corneal graft survival . A varieOver the last few years, substantial efforts have been made to understand ocular lymphatic and glymphatic systems, with a particular focus on their role in ocular pathophysiology. Findings from these studies contributed to our understanding of ocular fluid-drainage routes. It is generally considered that ocular fluid, particularly aqueous humour, drains via trabecular and uveoscleral pathways from the anterior segment of the eye . Previou


Flow cytometry was used to assess which immune cell populations infiltrate the tumours in response to the treatment combination. Imaging with [18F]AlF-NOTA-KCNA3P demonstrated that adjuvant metformin significantly improved anti-PD1 efficacy and led to a robust anti-tumour immunological memory response in a syngeneic colon cancer model through changes in tumour infiltrating effector memory T-cells.The low response rates associated with immune checkpoint inhibitor (ICI) use has led to a surge in research investigating adjuvant combination strategies in an attempt to enhance efficacy. Repurposing existing drugs as adjuvants accelerates the pace of cancer immune therapy research; however, many combinations exacerbate the immunogenic response elicited by ICIs and can lead to adverse immune-related events. Metformin, a widely used type 2 diabetes drug is an ideal candidate to repurpose as it has a good safety profile and studies suggest that metformin can modulate the tumour microenvironment, promoting a favourable environment for T cell activation but has no direct action on T cell activation on its own. In the current study we used PET imaging with [ V1.3 is overexpressed in active effector memory T-cells (TEM) [EM cells in responsive tumours versus non-responsive tumours [18F]AlF-NOTA-KCNA3P, a radiopharmaceutical specifically targeting KV1.3, is able to reproducibly identify lasting therapy response to ICIs [18F]AlF-NOTA-KCNA3P is sensitive enough to measure an enhanced immunological memory response induced by combining αPD1 with metformin.Immune checkpoint inhibitors (ICIs) have achieved great success in the field of immuno-oncology, but low response rates have prompted research to investigate combination strategies in an attempt to enhance efficacy ,2. Repurls (TEM) , and is ls (TEM)  and sign tumours . Recentl tumours FAlF-NOTAp = 0.881). The greatest response rate and tumour shrinkage was observed in the combined αPD1 plus Metformin treatment group, significantly greater than observed in the αPD1 monotherapy treated group, while metformin alone had no significant effect on tumour growth .A schematic representing the treatment regimen, tumour volume assessment and imaging for the animals is shown in r growth . The cri18F]AlF-NOTA-KCNA3P (on day 12) showed little tumour growth after re-challenge with tumour cells; however, TNR animals displaying low tumour retention of [18F]AlF-NOTA-KCNA3P (on day 12) showed significantly greater tumour growth after re-challenge . The control cohort, the metformin cohort and the TNR showed little [18F]AlF-NOTA-KCNA3P tumour retention, whereas the tumours responsive to αPD1 had significantly greater retention (** p < 0.01 compared to TNR). The αPD1 + metformin combination responders, however, showed even greater retention of [18F]AlF-NOTA-KCNA3P when compared to the TNRs (*** p < 0.001) and significantly increased retention compared to the αPD1 responsive group  were compared to tumours designated as treated non-responders (TNR) assessed using flow cytometry . ChangesEM cells .18F]AlF-NOTA-KCNA3P was isolated with a non-decay corrected radiochemical yield of 22.0 ± 6.4% within 50 min from delivery of aqueous [18F]fluoride. The radiochemical purity was greater than 99% and the molar activity was 32.5 ± 11.2 GBq/µmol at the end of synthesis  and radiolabeling was performed as previously described . [18F]Als n = 4, .5 cells per animal). The mice were dosed IP on days 6, 9 and 12 following tumour implantation with either 5 mg/kg rat IgG2a isotype control (α-trinitrophenol mAb) or 10 mg/kg rat IgG2a anti-mouse PD-1 . Metformin was dosed IP on days 6, 8, 10 and 12 following tumour implantation . Tumour volumes were measured longitudinally using calipers, and the tumour response was determined by measuring tumour growth inhibition as described in the All animal procedures adhered to the Singapore Institutional Animal Care and Use Committee regulations (IACUC No. 211649). The tumour implantation procedure was carried out as previously reported . Mice we18F]AlF-NOTA-KCNA3P tumour retention (>0.8%ID/g), and animals with tumours that were non-responsive to therapy (TNR) and low [18F]AlF-NOTA-KCNA3P tumour retention (<0.5%ID/g), were reimplanted with CT26 tumour cells in the contralateral left shoulder as described above. Tumour growth was assessed for a further 25 days.On day 22 post implantation, animals with tumours responsive to therapy (TR) and concomitant high [18F]AlF-NOTA-KCNA3P (~10 MBq) was injected via the lateral tail vein and tissue retention assessed using static PET imaging at 60 min post injection. Amide software  was used to analyse the static acquisitions and delineate volumes of interest to determine radioactivity retention in tissues. Data are expressed as % of the injected dose per gram (%ID/g).A Siemens Inveon PET-CT was used to image the animals at 12 days after tumour implantation, as described previously . [18F]Alhttps://github.com/JinmiaoChenLab/cytofkit, accessed on 2 May 2022). One thousand cells from each fcs file were used for analysis, using the following markers: KV1.3 (KV1.3 potassium channel marker), CCR7 (memory T cell marker), CD3 (pan-T cell marker), CD4 (helper T cell marker), CD8 (cytotoxic T cell marker), CD11b (myeloid marker), CD11c (dendritic cell marker), CD206, F4/80, I-A/I-E (MHC class II marker), Ly6C (macrophage marker), Ly6G (neutrophil marker), Nkp46 (pan-NK cell marker) and Siglec-F (eosinophil marker).Flow cytometry was used to assess the tumour-infiltrating immune cells as described in detail previously . The tump < 0.05 was considered statistically significant. Data are expressed as mean ± S.D. unless otherwise indicated.Kruskal Wallis 1-way ANOVA with Dunn’s post-test for multiple comparisons were used for statistical analysis of the non-parametric data sets . 18F]AlF-NOTA-KCNA3P reproducibly measured anti-tumour immunological memory T cell responses with increased retention, evident in tumours responding to PD-1 inhibition and significantly augmented after adjuvant metformin therapy. However, metformin had no effect on [18F]AlF-NOTA-KCNA3P retention alone  pathway during T-cell activation, and can lead to T-cell exhaustion AlF-NOTA-KCNA3P may prove effective clinically remains to be assessed. Tumour uptake of [18F]AlF-NOTA-KCNA3P is intratumoural, but may be affected by tissue necrosis or changes in vascularity, as KV1.3 is expressed in endothelial cells. Interpretation of tissue retention can be complicated by background uptake. KV1.3 has been shown to be expressed by some cancers AlF-NOTA-KCNA3P provides a non-invasive way to measure tumour infiltrating memory T-cells associated with metformin’s ability to enhance αPD1 response. Tumours exhibiting higher [18F]AlF-NOTA-KCNA3P retention demonstrated an on-going resistance to tumour cell re-challenge, suggestive of a durable anti-tumour memory response. The data indicate that [18F]AlF-NOTA-KCNA3P may be able to distinguish which drugs can be repurposed successfully as adjuvants to enhance the efficacy and durability of PD1 checkpoint inhibition clinically.Imaging with [