Body dissatisfaction is a public health issue, however, low awareness of its seriousness, and stigma, may inhibit treatment seeking. The current study evaluated engagement with videos promoting awareness of body dissatisfaction using a persuasive communication approach.n = 283) and women (n = 290) were randomly allocated to view one of five videos; (1) Narrative, (2) Narrative plus persuasive appeal, (3) Informational, (4) Informational plus persuasive appeal and (5) Persuasive appeal only. Engagement  was examined post-viewing.Men  were demonstrated for the persuasive appeal and informational videos relative to narrative approaches.Videos using clear and factual approaches may promote engagement in body image health promotion videos. Further work should be done to examine interest in such videos specific to men.The online version contains supplementary material available at 10.1186/s40359-023-01120-7. Body dissatisfaction, negative evaluation of one’s appearance, shape, or weight, poses a serious public health issue . It is aSocial marketing campaigns are becoming increasingly common within mental health –13, inclengaging. Social marketing engagement is a relatively recent area of research, and as such the literature is not yet extensive. Definitions of social media or social marketing engagement have typically come from research around brands. It is important to note that, within the context of this research and its applications, the term “brand” is applied broadly, and inclusive of non-government organisations and academic institutions, rather than solely focusing on capitalist consumer brands. While there is no one definition of engagement, there are commonalities in theoretical models of the construct [One essential component of social marketing campaigns is that they are onstruct –27. Modeonstruct  componenonstruct , 25. Theonstruct . As suchonstruct , and higonstruct , 30. Genonstruct , 32. As onstruct , 25, theThe type of communication that may result in greater engagement in a message has not been investigated within the context of body image. Previous research has identified two broad types of persuasive communication; narrative and informational , 34. InfThe effectiveness of both informational and narrative approaches has been found elsewhere throughout the literature with small effects on behavioural and attitudinal outcomes in meta-analyses , 36. It Persuasive health messages may also contain persuasive appeals, which are intended to clarify the content and meaning of narrative communication , 18, 39.While no literature exists examining the impacts of persuasive communication approaches on engagement, previous work suggests that narrative and informational approaches persuade using different mechanisms , 33, 42.Identifying whether particular domains of engagement may be particularly useful for targeted campaigns; for instance, campaigns aiming to promote socio-political change, may benefit from changing behavioural engagement, while campaigns aiming to reduce stigma may wish to impact cognitive and affective domains. As such, this study also sought to understand whether the type of persuasive communication may impact different engagement domains.Little research in body dissatisfaction and eating disorders has identified the specific approach taken in persuasive communications as either informational or narrative . In our As social media  relies heavily on sharing of information  and engagement , the current study sought to understand the effect of different types of persuasive communication on engagement in a body image social marketing video. Thus, the current study aimed to examine the extent to which such a campaign can generate cognitive, affective, and behavioural engagement and to compare levels of engagement across informational, narrative, and persuasive appeal communication approaches. As engagement has not previously been investigated within this context, no specific hypotheses were made. However, the following research questions were explored:RQ1: Of the persuasive approaches , which results in superior engagement in the topic of body dissatisfaction mental health literacy?RQ2: Do particular persuasive communication approaches impact the cognitive, affective, and behavioural domains of engagement differently?n = 24), failed the attention check (n = 3), were ineligible , or did not provide responses after watching the video (n = 25) that would exclude them from analyses, a final sample of N = 573  of approximately equal numbers of men  and women  remained. Participants ranged in age from 18 to 44 , and n = 7 not indicating their age. The majority of participants had one or more children (61.0%). The majority of participants resided in the United Kingdom (90.1%), followed by Canada (8.6%), and Australia (1.2%).Participants were recruited through Prolific, an online participant platform. Men and women aged 18–45 from Australia, Canada, or the United Kingdom were eligible to take part in the study. Participation was limited to persons from these countries as they are culturally similar to Australia, in which the stimulus materials were filmed. Initially, 633 participants responded to the study. After removing participants who chose to withdraw their data following debriefing where they learned the true aims of the study  to raise awareness of the seriousness of body dissatisfaction; one reflected a narrative approach to communication, one an informational approach, and the other was an explicit persuasive appeal. Experts in the field of body image and eating disorders gave feedback on the scripting and information prior to the production of the videos. Permission to use the materials in the research was granted by the media company.https://vimeo.com/entertainthinkinspire/tmp1.The narrative video (2.48 min) featured a woman in her early thirties engaging in an internal monologue of negative body talk  while driving in the car with her child. The woman eventually realises the harshness of her self-talk and acknowledges the functionality of her body over its appearance. The woman is white and thin, and was shown sitting. The actor was pregnant at the time of filming but this was not visible within the video as a bag was covering her torso. The narrative video can be viewed at https://vimeo.com/entertainthinkinspire/tmp5.The informational video (2.54 min) was comprised of interviews with five body image experts  who discussed factual information about body image  thin and b) women. One presenter was a larger bodied man. Only the upper bodies, not full-body view of presenters were shown. The video can be viewed at https://vimeo.com/156214950.The persuasive appeal video (1.20 min) featured the female actor from the narrative approach video presenting a direct appeal to the viewer to increase their awareness and understanding of body dissatisfaction, and question and challenge appearance ideals , as described above.The informational approach video was shown with footage edited in from the persuasive appeal video (3.45 min).Demographic questions included age, gender  to 100 (very much so). Previous research has demonstrated that scores from VAS are valid and reliable for assessing body satisfaction [A series of visual analogue scales (VAS) were used to assess weight satisfaction, shape satisfaction, and muscularity satisfaction before and after viewing the stimulus. Only pre-video exposure data around weight satisfaction, shape satisfaction, and muscularity satisfaction were used in the current analysis solely for the purposes of examining confounding effects, as the focus of the current study is on engagement outcomes. Participants were asked to indicate how satisfied they feel right now, from 0  = 4056.95, p < .001, suggested that there were sufficiently high correlations among items to perform EFA. Four components were identified with an eigenvalue of > 1, which explained 57.96% of the total variance. Parallel analysis using Vivek et al.’s [r = .01 to r = .36), and as such an orthogonal rotation was used. The rotated component matrix is displayed in the supplementary material  to 100 (very much so). A mean score from items responses was used for the scale score. Higher scores indicated greater interest. Cronbach’s alpha demonstrated excellent internal consistency, with α = 0.90.Eight VAS items identified from the factor analysis were used to measure participant interest  in the video and topic. Participants were asked to indicate their interest, from 0  to 100 (very much so). A mean score from items responses was used for the scale score. Cronbach’s alpha demonstrated acceptable internal consistency, with α = 0.62. Higher scores indicated greater compassion.Five VAS items identified from the factor analysis were used to measure participant compassion towards people with body dissatisfaction, as well as self-compassion. These VAS items measured affective engagement. Participants were asked to indicate the degree to which the video was respectful, increased other-directed compassion, oversimplified body image issues (reverse scored), increased blame (reverse scored), and was perceived to make other people feel more concerned about their appearance (reverse scored), from 0  to 100 (very much so). A mean score from items responses was used for the scale score. Cronbach’s alpha demonstrated acceptable internal consistency, with α = 0.69. Higher scores indicated greater relevance.Three items identified from the factor analysis were used to measure relevance of the video and topic, which reflected cognitive engagement. Participants were asked to indicate the degree to which the video was relevant to their own lives, covered an important topic, and they could recognise their own experiences in the video, from 0  to 5 (Strongly agree) about whether the video they watched was engagingTo ensure that the quality of the videos was reasonably similar across groups, participants were asked five questions, measured on a Likert-type scale from 1 ; as such, due to the potential for body satisfaction to confound the relationship between videos and engagement  this was controlled for in the analysis of relevance for women [p > .05), and as such two univariate ANOVAs were conducted to test whether videos produced differences in compassion and interest. All assumptions of ANOVA were met. For men, pre-existing muscularity, weight, and shape satisfaction were related to relevance. Thus, to test whether videos produced differences in relevance for men, an ANCOVA was conducted, controlling for pre-existing muscularity (p < .05), weight, and shape satisfaction (both p < .001) as potential confounds. Assumptions of ANCOVA were met. Body image variables were unrelated to interest or compassion for men , so two univariate ANOVAs were conducted to test whether videos produced differences in interest. All assumptions of ANOVA were met. Post-hoc power analyses for ANOVA and ANCOVA omnibus tests were conducted using G*Power [Analyses were separated by gender as women tend to have higher body dissatisfaction than men, which was verified using an independent samples t-test. Scores on all body satisfaction items were higher in men than in women; all or women . Thus, t G*Power , which rps < 0.001). The narrative and narrative + persuasive appeal videos were significantly more humorous than all other conditions (ps < 0.001). These findings suggest the manipulation was successful. The production quality was rated significantly higher in the narrative condition, the information condition, and information + persuasive appeal than the persuasive appeal condition (p < .001). Additionally, the visual appeal was rated as higher in the narrative only video, the information + persuasive appeal and the narrative + persuasive appeal than the persuasive appeal video .To ensure that the quality of the videos was reasonably similar, we examined participants’ responses to the manipulation check questions. A series of one-way ANOVAs examining the differences between the five groups on the manipulation check questions were conducted. Holm corrections were used to account for multiple comparisons. The information and information + persuasive appeal videos were rated more factual than both the narrative and narrative + persuasive appeal videos  = 1.22, p = .30, partial η2 = 0.02.As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to interest in women, a univariate ANOVA was conducted to examine the differences between video conditions on interest. There was no significant effect of video condition on interest, F  = 42.38, p < .001, partial η2 = 0.13), and shape satisfaction  = 0.9.34, p = .002, partial η2 = 0.03) were significantly related to relevance; but muscularity satisfaction  = 2.39, p = .12, partial η2 = 0.01) was not. There was no significant effect of video condition on relevance, F  = 1.61, p = .17, partial η2 = 0.02.As measures of pre-test weight satisfaction, shape satisfaction, and muscularity satisfaction were related to relevance in women, an ANCOVA controlling for these variables was conducted to examine the differences between video conditions on relevance. Pre-test weight satisfaction  = 4.67, p = .001, η2 = 0.06. Holm corrections revealed that the persuasive appeal video  resulted in significantly greater compassion than the narrative only video , adjusted p = .002. The information only video  resulted in significantly greater compassion than the narrative only video , adjusted p = .005. No other significant differences were found.As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to compassion in women, univariate ANOVA was conducted to examine the differences between video conditions on compassion. There was a significant effect of video condition on compassion, All descriptive statistics for ANOVAs and ANCOVA analyses for men examining the differences in engagement by video condition are displayed in Table F  = 0.66, p = .62, η2 = 0.01.As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to interest in men, a univariate ANOVA was conducted to examine the differences between video conditions on interest. There was no significant effect of video condition on interest, F  = 7.44, p = .007, partial η2 = 0.03, but weight satisfaction  = 0.34, p = .56, partial η2 < 0.01) and muscularity satisfaction  = 1.96, p = .16, partial η2 = 0.01) were not. There was a significant effect of video condition on relevance, F  = 4.45, p = .002, partial η2 = 0.06.As measures of pre-test weight satisfaction, shape satisfaction, and muscularity satisfaction were related to relevance in men, an ANCOVA controlling for these variables was conducted to examine the differences between video conditions on relevance. Pre-test shape satisfaction was significantly related to relevance; p = .005) and the narrative + persuasive appeal video . The persuasive appeal  resulted in significantly greater relevance than the narrative only , and the narrative + persuasive appeal video . The information + persuasive appeal video  resulted in significantly greater relevance than the narrative only video . No other significant differences were found.Holm corrections revealed that the information only video  resulted in significantly greater relevance than the narrative only video  = 3.57, p = .007, partial η2 = 0.05. Holm corrections revealed that the information and persuasive appeal , information only , and persuasive appeals  all resulted in significantly greater compassion than the narrative only video . No other significant differences were found.As measures of pre-test muscularity, weight, and shape dissatisfaction were unrelated to compassion in men, a univariate ANOVA was conducted to examine the differences between video conditions on compassion. There was a significant effect of video condition on compassion, The aim of this study was to compare participants’ reported engagement with different forms of persuasive communication about body dissatisfaction, focusing on informational, narrative, and direct persuasive appeal. We proposed two research questions. The first was whether a particular persuasive approach resulted in superior engagement over other approaches in the context of body dissatisfaction mental health literacy. It was found that there was some advantage for both persuasive appeals and information-only approaches over a narrative approach for both gender groups, and some advantage for the information approach with the addition of a persuasive appeal for men. The second was to understand whether different persuasive communication approaches impacted cognitive, affective, and behavioural domains of engagement differently. It was found that there were some domains impacted differently depending on the communication approach, in particular for cognitive and affective domains.Overall, while all communication types were engaging, there appeared to be some advantage for persuasive appeals and informational videos across both men and women. This is only partially consistent with the previous literature around persuasive communication, which has found benefits overall for both informational and narrative approaches , 36. AltThe benefits observed from viewing a persuasive appeal (relative to the narrative videos) are a novel finding, as to our knowledge the effects of viewing a standalone persuasive appeal were previously unexamined in the literature. This finding may indicate that the nature of a persuasive appeal is appropriate for use on social media or in other forms of media. The message is short, clear, and action-based, giving viewers a specific call to action to learn more about body dissatisfaction as an important mental health issue. This appeal bears resemblance to very successful strategies commonly used in social media  which are intended to generate engagement . It is iDifferent impacts on domains of engagement were found, which were partially consistent with literature on processing communication , 44. IntInconsistent with theories of processing and attitude bases , 44, 60,Notably, although short videos showed differences for compassion and relevance (for men) and compassion (for women), no videos appeared to produce superior results in all domains of engagement across genders. This finding is not especially concerning in itself, particularly when examining the findings for relevance and interest in women . It should be of note, however, that interest  was moderate among men, which may indicate less intent to engage with body image content more generally as body dissatisfaction and eating disorders are perceived to be feminine issues . ReasonsThe findings of this study have implications for social marketing campaigns within the body image field, and research into their efficacy. First, it is clear that further research exploring the impact of persuasive appeals should be conducted, as they were more engaging than narrative approaches within some contexts. Second, contextualising the current study’s findings with our previous work, social marketing campaigns that aim to educate and promote awareness through engagement may take a different approach from those aimed at improving body satisfaction. There may be some advantages to informational and persuasive appeals relative to narrative approaches within the context of engagement. Campaigns may wish to use expert information, or direct appeals to the audience, particularly to generate interest online. Third, research should further examine the impact of particular persuasive communication approaches on domains of engagement. For instance, a call to action may best be used to impact affective engagement. Further examination of persuasive communication approaches on specific domains of engagement would be beneficial for social marketers and researchers.intended behaviour rather than actual behaviour, and it would be beneficial to evaluate such messages including a measure of actual behaviour – for example, whether participants click on a link to visit a website about body image.A strength of this study was its generalisability; a mixed-age, community sample of men and women was used, which is not common in evaluating social marketing or anti-stigma campaigns for body image and eating disorders . The meaFurther limitations were that engagement was examined at one point in time, which precluded examination of change in engagement resulting from video viewing, and that the use for the cover story likely did not fully conceal the purpose of the study. In addition, the standalone persuasive appeal video was rated as less visually appealing and with lower production quality than the other videos, which may have impacted interest and attention to the videos. Another limitation is the varying length of the videos, in particular the difference in viewing time between the persuasive appeal only video, and the conditions to which the persuasive appeal video was added. As such, the effects of video length from video content may be difficult to separate. Although all videos were short in duration, future research should consider using equivalent length videos in all conditions. An additional limitation is around cultural relevance; although we collected country of residence, we did not collect information on other demographic characteristics; which may be of relevance considering academics and actors appearing in the videos were white. Future research examining engagement in such videos may wish to collect information such as cultural background, to better assess generalisability of the research, and aim to ensure the cast is not all or majority white to better reflect a community sample’s characteristics. Finally, a limitation is that most people in the video were thin; this may have unintentionally reinforced the idea that body dissatisfaction is only a problem when the person’s body is smaller; further research should examine the impact and potential benefits of educating the public that body dissatisfaction has a negative impact at all sizes, the issue of weight stigma, and attempt to include more size diversity.The current study demonstrated that attempts to raise awareness of body dissatisfaction through persuasive communication were highly engaging in relation to behavioural (interest), affective (compassion) and cognitive (relevance) domains. Although all videos were rated as highly engaging, persuasive appeals and informational approaches were rated more highly on some engagement domains by men and by women. These findings suggest that video messages that are demonstrated to be effective in increasing perceptions of the seriousness of body dissatisfaction, as our previous research demonstrated , may alsBelow is the link to the electronic supplementary material.Supplementary Table S1 and S2


Less is known about non-Western social media use and its effects on body image concerns. Chinese TikTok, known as Douyin, is a popular short video platform with 600 million daily active users. Recent trends on Douyin encourage users to demonstrate thinness through participation in ‘body challenges’. This paper argues that such content is comparable to thinspiration, however, to date hardly any research has been undertaken on these challenges. Thus, this pilot study aimed to analyse the content of three viral challenges and investigate their impact on Douyin users.Social media content on Western platforms promoting thinness, or Thirty most viewed videos were collected for three challenges (N = 90): the Coin challenge, the A4 Waist challenge, and the Spider leg challenge. Videos were coded for variables relating to thin idealisation, including thin praise, sexualisation and objectification, and analysed through content analytic methods. Video comments (N ≈ 5500) were analysed through thematic analysis, and main themes were identified.Preliminary findings showed that participants who objectified their bodies to a greater extent expressed more negative body image concerns. In addition, comments on the videos had themes of thin praise, self-comparison, and promotion of dieting behaviours. In particular, videos of the A4 Waist challenge were found to incite more negative self-comparison in viewers.Preliminary findings suggest all three challenges promote the thin ideal and encourage body image concerns. Further research about the broader impact of body challenges is needed. Social media plays an important role in shaping public perceptions of body ideals. Short-video platforms, such as TikTok, have further facilitated the dissemination and consumption of media content and popularised challenge videos that involve wide user participation. The aim of our study was to analyse the content of three viral Chinese TikTok (Douyin) body challenges  and investigate their impact on Douyin users. We extracted 90 short videos featuring the three body challenges from Douyin using Python and up to 100 comments from each video for qualitative analysis. Our findings indicated that the overt thin-ideal messaging, objectification and sexualisation of bodies in these body challenges may contribute to upward appearance comparison and greater body dissatisfaction. The present study provided initial support for the detrimental effects of Douyin body challenges on body image for both viewers and content creators. Through this pilot study, we hope to inform further research on the relationship between appearance-oriented content on short-video platforms and body image concerns.The use of Western social media sites such as Facebook, Instagram, and Twitter have been linked to body dissatisfaction and disordered eating in users globally –4. GreatCompared to traditional mass media, social media has been found to have a pronounced impact on body dissatisfaction as it encourages upwards appearance comparison (comparing oneself to a body perceived to be more attractive than one’s own) with both peers and celebrities compared to the previous mass media that mainly focused on celebrities , 9. The Importantly, following the outbreak of COVID-19, researchers have theorised that an increased use of social media during the pandemic and concerns about health and fitness during COVID-19-related isolation may have increased the risk of developing eating disorders in vulnerable individuals , 11. In Given the prevalence of social media use globally, with an estimated 4.26 billion users in 2021 , it is iStudies focusing on a Chinese population have found that traditional mass media reflects societal expectations to be thin, which creates perceived appearance pressure that is linked to body dissatisfaction and other risk factors (such as negative affect) for disordered eating among young women –23. SpecIn the past decade, the rising trend of online videos encouraging viewers to undertake or participate in ‘body challenges’ showcasing users' thinness has drawn concern about their contribution to increased disordered eating behaviours . These vDespite implications of the challenges on body dissatisfaction and disordered eating, it appears that only one study has investigated the impacts of Douyin challenges on users’ body dissatisfaction. Jackson et al.  examinedthinspiration and fitspiration content on Western platforms including YouTube, Instagram, Tumblr, and Twitter ’.每天睡前练一遍, 你的肚子腰线就会越来越漂亮了。  - A4坚持30天甩掉10斤小赘肉! (Keep it up for 30 days to lose 10 pounds of muffin top fat!) - A6暑假大吃特吃, 然后肚子越来越大, 输家(sic)前买的衣服都穿不上了  - A30Several subjects presented informative videos on a range of eating restriction, exercise and weight loss advice either directly to succeed in the challenge or included the hashtag to leverage the challenge’s popularity to increase personal exposure. Interestingly, all mentions of eating restriction were not voiced by the video subject themselves but were present in the voiceovers.明天李现绯闻女友, 你来当。  - A3为啥你的腰粗想个大水缸? (Why does your stomach look like a big water pot?) - A18男朋友更爱我了 (Your boyfriend will love you even more) - A30Many videos explicitly stated reasons for idealising thinness, including health, romantic success. Some commended certain features of thinness such as a bony appearance or denounced fat.Figure @user 我想看你拍个这个 (@user I want to see you film this) - SL20.92Many comments expressed their approval of the challenge itself, seeing it as an instrument worthy of being spread and popularised. The most common type of comment was a user tagging another user.There were several notable subsets within praising video subjects. Approval of the subject’s physical traits was frequently observed. Another common method of praising video subjects was upward comparison, originally described by Festinger (1954). A small number of comments praised the subject’s non-physical attributes such as humour or personality.你的身材是我的目标 。I envy your body figure) - SL12.40我(...马达啦...)愿称你为蜘蛛腿系列最美, 腿好长呀。  - SL5.1个人喜欢王丽坤胯宽腰细才好看  - A1.96看完你所有的视频。我决定了吃完这顿再考虑减不减肥这问题  - A12.82我就想问问吃什么小腿可以比躯干长?  - SL6.34小腿肚子都是肌肉怎么能瘦一点  - SL29.29A subset of praise was indirect, taking the form of upwards comparison, which was often paired with a desire to restrict eating, or seek advice to lose weight.这是传播的什么文化? (What type of culture is this spreading?) - SL9.6新时代裹小脚 (Modern day foot-binding) - A1.51都在挑战这个动作, 可是这个动作又不难。  - SL26.11Comments conveying criticism of the challenge expressed concern for potential impacts of the challenge on society. However, another more prominent type of critical comments often belittled the value or difficulty of the challenge.抱歉, 姐姐, 我从来都没见过腿这么短的蜘蛛。  - SL4.30Similar to praising the video subject, many comments criticising the video subject equally mentioned their physical traits. Many drew comparisons between the subject and an object, or other people.The present study aimed to investigate the influence of three trending Douyin challenges  on body dissatisfaction, body image concerns, and disordered eating behaviours of social media users. The main findings of the content analysis suggest that, similar to thinspiration and fitspiration content on Western social media platforms , 39, DouIn order to better present the uniqueness of Douyin, variables mentioning celebrities and challenging others were created as a new addition to the codebooks from previous literature, which included a range of descriptive attributes, appearance, body type and objectification, thin-ideal messages, and comparison, and mentions of food and exercise , 39–41.From the content analysis of the 90 Douyin videos, the predominant body type of video subjects was the thin body type across all three challenge videos. Video subjects who had a thin body type were present in 93% of the Spider leg challenge videos, 73% of the Coin challenge videos, and 70% of the A4 Waist challenge videos. More video subjects had thin body type in the Spider leg challenge videos as opposed to the other two challenges, which may be due to the nature of the Spider leg challenge that required participants to film their entire bodies . The overall predominance of thin bodies in the videos aligns with findings from studies which performed content analysis on fitspiration and thinspiration content on Western social media sites and suggested that the Western female body ideal is one of thinness , 36, 43.This suggests that the Douyin body challenge videos, as a form of social media designed to show off video subjects’ physical appearances, may enable appearance comparison in a similar manner to thinspiration and fitspiration content. Social comparison has been theorised as the major mechanism for media exposure to lead to body dissatisfaction and disordered eating through sociocultural exposures . The mosOur findings also align with the only experimental findings of the impact of Douyin challenges on body dissatisfaction , which fSome proportion of the body challenge videos contained thin praise. Thin praise was found in 50% of the A4 Waist, 16.67% of the Coin and 3.33% of the Spider leg challenge videos. As above, this draws similarities with thinspiration and fitspiration images of women on Western social media sites. In particular, Tiggemann and Zaccardo  found thThe discrepancy between frequency of thin praise in each of the challenges  may be explained by the nature of the challenges. As the Spider leg challenge required participants to perform a difficult pose (squatting like a spider), video subjects tended to speak more about the flexibility and physical ability to ‘succeed’ in the challenge. Thus, the focus was not solely on the physical appearance of the participants, as thinness itself was not sufficient to succeed. In comparison, the A4 Waist and Coin challenges only required participants to showcase their physical appearance, which may explain the higher frequency of thin praise in those challenge videos. However, despite the additional emphasis on participants’ physical ability in the Spider leg challenge, Spider leg videos still had the highest percentage of thin video subjects out of the three challenges , even though ostensibly, thinness was not required to succeed. This may suggest that despite the superficial emphasis on physical ability, video subjects in the Spider leg challenge were primarily focused on their appearance and chose to participate to showcase their thinness.The notion of displaying physical thinness through participating in the challenges is further bolstered by the findings relating to Thin Pose. A high proportion of video subjects in the A4 Waist challenge (80%) performed a thin pose. This finding is expected, as the A4 Waist challenge required participants to showcase their physical thinness. However, in the Coin challenge, almost half (43.33%) of video subjects performed a thin pose, although the challenge ostensibly did not require physical thinness to succeed. This may suggest that Coin challenge participants were conscious of their physical appearance while undertaking the challenge. bonespiration, a subset of thinspiration images glorifying bone protrusions found on Western social media sites such as Instagram, Tumblr and Twitter [Bone Emphasis was specific to the Coin challenge, given that the focus of this challenge was on one’s collarbones. Hence, this behaviour was found in all of the Coin challenge videos and did not appear in the videos of the other two challenges. This suggests that there is a strong similarity between the Coin challenge videos and  Twitter . Hence, Mentions of exercise were coded in 20% of the A4 Waist, 6.67% of the Coin, and 3.33% of the Spider leg challenge videos. Research has consistently reported that exercise decreased body dissatisfaction and reduced body image concerns , 60. HowCompared to the Coin and Spider leg challenge videos, a greater proportion of A4 Waist challenge videos were coded as sexualising or objectifying the video subjects. Sexualisation variables were coded the highest in the A4 Waist challenge, compared to the Spider leg and the Coin challenge. Overall, tight clothing was found in 41.11% of all videos collected across the three challenges, which became the most represented sexualisation variable in this study. This also draws similarities with thinspiration and fitspiration content with findings from Alberga, Withnell , which fObjectification can influence the degree of sexualisation, which ultimately affects body image concerns. We theorised that body focused Douyin challenges would have greater objectification elements, and objectification of video subjects was approximated by the incidences of videos which did not show the full body of the subject. While the Spider leg challenge videos tended to be less objectifying in the sense that more of the body was shown (100% of videos showed the full body), most of them (60%) deliberately obscured the subject’s face or eyes. As the Coin challenge videos emphasised the collarbones, all of them did not show below the subject’s torso. Although the Coin challenge videos were coded as the most objectifying , this may be due to the nature of the challenge’s focus on collarbones. This is similar to findings from thinspiration studies, which found that Western thinspiration content tended to be objectifying by only focusing on specific parts of the body . In agreThe content analysis results indicated that the variables ‘Mentioning Mental Illness’ and ‘Dieting Restraint Messages’ were not found in the challenge videos, indicating that video subjects did not verbalise sentiments relating to mental illness or eating restriction in the videos. This was surprising given that previous literature has consistently reported positive correlations between internet use, body image concerns, restriction of eating and eating pathology , 63. It Besides eating restriction, the mention of mental illness was also absent in all of the videos collected. The lack of mention for mental illness may be the result of cultural differences. Mental health is highly stigmatised in Chinese culture, and it is closely correlated with moral values and societal expectations , 69. A pMention of celebrities was coded in 6.67% of the A4 Waist, 6.67% of the Coin, and 3.33% of the Spider leg challenge videos. Even though mention of celebrity was not coded in many of the challenge videos, it was still important to include this variable because of Douyin’s unique operation system linking video subjects’ profit or other positive outcomes, such as fame, exposure, sponsorships, etc., with the popularity of their videos. By mentioning trendy celebrities, the video subjects are not only bringing the attention of celebrities’ fans to the videos but also garner views through relevant trends and potentially increasing their own fans . For exaChallenging others was the other new variable for this study and was coded in 26.67% of the Coin, 6.67% of the Spider leg, and none of the A4 Waist challenge videos. Although not all of the challenge videos collected had challenged others, it was included as it is also unique to the nature of a Douyin Challenge. Video subjects use challenging others as a strategy to encourage engagements from the viewers and interact with the viewers in the hope to gain more impact on the Douyin platform. The operation system of Douyin allows the video subjects and viewers to interact in a way where viewers are presented with content that they are interested in, and video subjects are gaining profit or popularity through the engagement  of their viewers . This opIn addition, the challenges often encourage people to use the same music and the same filters to maintain the consistency of a challenge, which could play a role in normalisation of appearance-changing filter use. In fact, 96.6% of the Spider leg, 40% of the A4 Waist and 40% of the Coin challenge videos respectively were found to use the same music. Similar filters within the videos collected for each challenge were also found in 86.67% of the Coin, 70% of the A4 Waist, and 60% of the Spider leg challenge. When challenge videos use the same music and dramatic filters, it is difficult to differentiate whether the thinness of participants is attributed to their natural body or a filter that makes them look consistently thin and long-legged. Furthermore, the use of appearance-changing filters was found in the videos across the three challenges, as some of the videos seemed to be purposely blurry and colour corrected, which could be a way to cover blemishes and change appearance. With increased filter use associated with the challenges, users are increasingly exposed to a falsely thin and polished appearance, which is dangerous as these false images are becoming normalised and providing an unattainable benchmark for a normal body type.Challenging others were coded in 26.67% of the Coin, 6.67% of the Spider leg, and 0% of the A4 Waist challenge videos. A video subject could challenge their viewers to engage in the activities verbally in the videos, by using captions, and by using hashtags. We theorised that whether and how a video subject decides to challenge their users depends on the nature of a challenge. If a challenge was more popular on social media platforms due to celebrities, TV shows, or other reasons, then video creators and subjects may be more likely to use challenge others as a strategy to stay on-trend and relevant. This theory could explain why challenging others was coded the highest in the Coin challenge videos, as many celebrities have participated in this challenge on TV shows. It is unclear why challenging others was coded in none of the A4 Waist challenge videos. A possible explanation may be that the A4 Waist challenge is an older challenge from 2016 and people have eventually moved on from actively participating in this challenge at the time of video extraction. Other factors that may affect the coding of challenging others need to be investigated. We theorised that potential research factors may include: (1) the age groups or populations that a challenge is targeted to, (2) the impact factor  of the celebrities who participated in a challenge, (3) the impact factor of the media platforms, (4) where the challenge is showcased, and (5) cultural factors that impact the value of body/beauty traits promoted by a challenge.The audio transcripts as well as video captions and text provided a deeper insight into the behaviours of video creators, beyond the visual content of subjects, as some creators offered commentary while including content of others. Creators focused heavily on appearance ideals, illustrated in the subthemes Challenge as a Reflection of Ideal, Collective Struggle to Achieve Ideal, Methods to Achieve Ideal, Thin is Good, and common references to Muscular and Posture. The challenges provided a benchmark of ideal appearance, equating success with achievement of beauty standards. Fans want popular subjects to participate in the challenges to prove their beauty (Pressure to Participate), and many subjects express happiness in succeeding (I’m Happy I Succeeded), an indication that they have attained the benchmark. The most prominent appearance ideal mentioned is thinness, which is further glorified by being linked to romantic success and health. These ‘positive’ connotations are reinforced with the results of the content analysis, with mostly thin subjects objectifying their bodies and portraying suggestive connotations.As noted above, a higher proportion of the A4 Waist challenge videos contained voiceover content by the video creators, not the speech spoken by the video subjects themselves. Accordingly, there was a higher proportion of exercise videos under the A4 Waist challenge. Several of these videos provided educational guides from supposedly certified doctors or personal trainers to help viewers lose weight and to succeed in the challenges. This reflects the findings of Ratwatte and Mattacola  highlighTo conclude, the content analysis described above demonstrated that most subjects were coded as thin, a small number were coded as average, but none had larger bodies or were coded as overweight. Combined with their themes of encouraging everyone to complete exercises together and lose weight , they portray that challenge success is more easily achieved. They also limit the range of body types that are idealised, which is shared with thinspiration and fitspiration studies , 39, andThematic analysis of comments provided an insight into the impact of challenge videos on viewers. The four major themes of the video comments were Praise of Challenge, Praise of Video Subject, Criticism of Challenge, Criticism of Video Subject. More comments were found to fit the theme Praise of Challenge than Criticism of Challenge. Those who criticised expressed concern about the impact of the challenge on culture. However, the most frequent comments across all three challenges were tagging and challenging others. In comparison, previous thematic analysis studies on thinspiration and fitspiration did not note any subjects or commenters actively challenging other users , 37. FurMany commenters engaged in negative self-comparison. Commenters directly expressed a desire to diet after watching such videos, and many sought exercise advice, which is in line with many previous studies suggesting that repeated exposure by social media to unrealistic ideals of beauty in females is a risk factor for body dissatisfaction, weight concern, thin-ideal internalisation and disordered eating behaviours in women , 5. ThisWithin both the themes Praise and Criticism of Video Subject, there were more comments about Physical Traits of the subject compared to their Non-Physical Traits, suggesting the nature of the challenges place greater fixations on certain body parts, which reflects the findings of our content analysis of high levels of objectification in the videos. In accordance with this finding, an experimental study by Tiggemann and Barbato  looking While no studies directly investigate the effects of negative social media comments on video subjects, the Tripartite Influence Model  providesThe current study on Douyin challenges has several theoretical and clinical implications that are important to outline in hope to inform public health and guide future research studies.First, this study has added unique cultural values to the theoretical association of social media use and body image concerns. Research has consistently reported body dissatisfaction being a strong risk factor for eating pathology, eating restriction, and the onset of eating disorders , 48. TheThe social expectations and peer pressure to look a certain way as everyone else may also be a result of such unique cultural values. In addition, it has also been reported that tags or language used in pro-eating disorder communities have constantly been deleted and removed to protect vulnerable individuals, which facilitates the evolution of language used to refer to certain pro-eating disorder content over time . The lanClinically, developing body dissatisfaction because of cultural values suggests that Asian populations are as vulnerable to unhealthy body-focused trends and as susceptible to developing eating disorders compared to other populations. As implicated, attention should be focused on not only Chinese populations, but also other Asian countries or cultures that may also share similar collectivistic cultural values as Chinese culture. For example, Mukbangs, eating shows that originated in South Korea where video subjects consume large amounts of food on camera, have been reported to be associated with restrictive eating and loss-of-control eating, and overall mental health problems , 82. ForConsidering the above, to prevent the worsening of body image concerns, disordered eating behaviours, and new onset of eating disorders, it may be helpful to monitor trends on social media and short-video platforms that are body-focused. Our current study also addressed special considerations when assessing the associations of social media use and body image concerns in Asian population. It would be important to teach the users to recognize cultural differences, as some cultural values may also interact with the association of social media trends and body image concerns. In addition, there is a need for social media literacy training, which includes teaching users that the body of video subjects in these challenge videos may be purposefully altered to meet unrealistic body expectations.The study was affected by several limitations, which are important to outline. The primary limitations of this study include the cultural context of the coders and the limited sample size of videos and comments extracted in the content analysis. First, the cultural context of each of the coders for both the content and thematic analyses informed their coding decisions, which may misrepresent the sentiments expressed by video subjects and commenters. While the three coders  are of Chinese heritage and have competency in the Chinese language, they are not currently located in China and received some parts of their education in Australia and the United States. Every effort was made to interpret meaning through a Chinese cultural lens; however, it must still be noted that the thematic and content analyses were conducted by coders with a Western-biased understanding of social media use, landscape, and lifestyle integration.Second, the relatively small sample size of videos (N = 90) and comments extracted (N = 5508) is another limitation in that the coded videos and comments may not be an accurate representation of the population of videos and may not reflect the trend at large. It is critical to note that many of the variables outlined in Table Lastly, a final limitation was potential errors in extracting the comments. Python was used to extract the comments into an Excel file format, the visual impact of some emojis were limited.Despite these limitations, the study also had several notable strengths. Its primary strength was the application of an inductive, mixed methods approach to the analysis of a novel set of data . Specifically, where literature is scarce, the use of inductive research methods allowed the data to guide research hypotheses without preconceived biases. In addition, the study was able to consider both Chinese and Western cultural contexts comparatively due to the coders’ language competencies in both English and Chinese. Furthermore, analysis of comments demonstrated it can be a rich source for investigating the impacts of social media content on viewers.This study aimed to explore the relationships between Douyin body challenge videos and body dissatisfaction in both video creators and viewers. Overall, the trending challenges contain content of concern with regards to body image and risks for disordered eating. The nature of the challenges, elements of objectification and sexualisation, thin-ideal messaging and unnatural filtered appearances contribute to negative and problematic attitudes towards body image. Thematic analysis of users’ comments revealed that viewing challenges can lead to greater body dissatisfaction, in addition, viewers can participate and objectively compare themselves. Moreover, the widespread nature of challenge through users’ participation and tagging, or digitally by the Douyin platform itself, means the risk of body dissatisfaction or disordered eating may be greater than other social media trends with similar risks. The study hopes to inform further research on short-video platforms similar to Douyin, including TikTok or YouTube Shorts or Instagram Reels, or Chinese social media, or challenge videos specifically.


Background: Midline exploratory laparotomy is essential in emergency surgery, and effective closure of the abdominal wall is crucial for optimal healing and reduced complications. The far-near-near-far technique for rectus sheath closure has gained attention due to potential advantages over conventional closure due to the fact that even if one suture gives way it does not affect the nearby suture and the rectus sheath is still held in place. This study aims to compare these techniques in emergency exploratory laparotomy.Methods: This single-center prospective observational study included all patients undergoing emergency exploratory laparotomy at a tertiary care hospital. Patients were categorized into two groups based on the closure technique used: conventional closure or far-near-near-far technique. Sociodemographic data and comorbidities were collected. Operative time, closure time, and postoperative complications were assessed. Statistical analysis was performed using IBM SPSS Statistics 22.0 .Results: The study included patients with a mean age of 42.14 years. Operative and closure times did not significantly differ between the groups. There was no significant difference in comorbidities between the two techniques. The incidence of wound infection, dehiscence, burst abdomen, incisional hernia, and sinus formation did not significantly differ between the groups. Late complications were also similar.Conclusion: The far-near-near-far technique for rectus sheath closure in emergency exploratory laparotomy showed comparable outcomes to conventional closure methods. No significant differences were found in operative time, closure time, or postoperative complications. This study contributes to the understanding of different closure techniques, allowing surgeons to make informed decisions In emergency surgery, midline exploratory laparotomy is crucial in diagnosing and treating a wide range of life-threatening abdominal conditions . The sucConventionally, abdominal wall closure involves a continuous suture technique or interrupted sutures placed in a layered fashion. However, concerns have been raised regarding these closures' strength, integrity, and durability, particularly in emergencies where the abdominal wall is often under increased tension due to trauma or infection. In contrast, the far-near-near-far technique offers an innovative approach that incorporates multiple sutures and distributes the tension more evenly, potentially minimizing the risk of wound dehiscence and incisional hernias . A meta-This clinical comparison study aims to evaluate the outcomes associated with the far-near-near-far technique in comparison to conventional closure methods. Parameters such as time taken to operate, post-operative abdominal wall complications such as wound infection, dehiscence, burst abdomen, incisional hernia rates, and sinus formation were examined to assess the effectiveness of the far-near-near-far technique in an emergency exploratory laparotomy. Understanding the nuances of different closure techniques is vital for surgeons, enabling them to make informed decisions and employ the most appropriate method for each patient . By shedStudy designIt is a single-center prospective observational study conducted at a tertiary care hospital, AIIMS Bhopal.Study settingThis study was conducted in the surgery and emergency department at All India Institute of Medical Sciences, Bhopal a tertiary care teaching hospital in Central India. The study was conducted from March 11, 2020 to December 2021.Study populationAll patients over 18 years of age undergoing emergency exploratory laparotomy procedures at AIIMS Bhopal between March 2020 to December 2021 were included in the study. Patients who underwent elective laparotomy, pre-operative proven malignancy, previous laparotomy for any condition , discharge against medical advice or death before the 10th post-operative day, any requirement of a re-laparotomy within 10 days for a cause not attributable to a complication of wound closure and lack of adequate post-operative follow-up were excluded from the study.Sample sizeThis study incorporated universal sampling and included all the patients who underwent emergency exploratory laparotomy surgery in the surgery and emergency department at AIIMS Bhopal.Ethical statementPermission from the Institutional Human Ethics Committee- Post Graduate Research (IHEC-PGR) (LOP No: IHECPGRMD047) was taken before the commencement of the study. The subjects were informed about the research and procedures through a participant information sheet. Data collection was not done without written informed consent. Confidentiality of information has been maintained. Only the research team has access to personal information. Personal identifiers were removed during data analysis and reporting.Data collectionPatients undergoing emergency exploratory laparotomy surgery from March 2020 to December 2021 who fulfilled the inclusion criteria were approached and informed consent was taken. Patients were included in two groups according to the surgeon's closure method, i.e., conventional closure or “far-near-near-far” technique of abdominal wall closure. An instrument was developed to obtain the demographic and clinical data of the subjects in the study. The instrument consisted of two parts: Demographic and Clinical data. The demographic data included the following information: age, sex, date of admission, date of operation, date of discharge, number of post-operative outpatient visits, and period of postoperative follow-up in the hospital. Clinical data included information about the patients’ health status: the presence of comorbid illness, lifestyle factors, the diagnosis of the patient, the focus of insult, the operation done, the stoma formed, admission in the surgical intensive care unit for monitoring/mechanical ventilation, postoperative complications and the period of follow-up in the out-patient department. The follow-up was for twenty-one months and outcomes included wound infection, wound dehiscence, burst abdomen, and late complications .Operational definitionsContinuous/Conventional ClosureConventional closure included closure of rectus fascia in a continuous fashion. The sutures were placed 1 cm from the edge of the Linea alba on both sides and a 1-cm distance was maintained between two adjacent sutures.Far-Near-Near-Far/Interrupted ClosureThis technique included suture approximation of the rectus sheath, in an interrupted fashion. The entry and exit of Polypropylene was 2 cm from the wound edges and 1 cm from the edge of Linea Alba on either side. The distance between two adjacent sutures was 1 cm.Statistical analysisStatistical analysis was carried out using statistical packages for IBM SPSS Statistics 22.0 for Windows . Continuous and categorical variables were expressed as frequencies, means, and percentages, respectively. The chi-square test was used to compare categorical variables. Two-sided p-values were considered statistically significant at p<0.05. Logistic regression was done to assess the association of demographic variables, comorbidities, and lifestyle factors with wound infection and wound dehiscence.The flow of the study is depicted in Figure The study revealed that the mean age of the subjects enrolled was 42.14 ± 15.12 years. The majority of subjects in the study were obese and males comprised 68.25% of the study population. Table The mean total operating time in the continuous group was 177.25 ± 45.88 minutes which was longer than that in the far-near-near-far group was 160.80 ± 45.27 minutes (p=0.157). The mean total time to close the wound in the continuous group was 16.34 ± 3.219 minutes which was slightly lesser than that in the far-near-near-far group 17.94 ± 3.54 minutes (p=0.067). The two groups matched in terms of comorbidities and lifestyle factors. 18.8% patients in the continuous group had a stoma made and required post-operative ICU admission in contrast to 41.9% patients in the far-near-near-far group (Table Table Table The closure of the rectus sheath is critical in exploratory laparotomy, particularly in emergencies where the abdominal wall is under increased tension. The choice of closure technique can significantly impact patient outcomes, including the incidence of postoperative complications . In thisOne of the far-near-far technique's key advantages is its ability to distribute tension more evenly across the abdominal wall, potentially reducing the risk of wound dehiscence and incisional hernias. The far-near-near-far technique of abdominal wall closure is advantageous because it prevents the simultaneous involvement of the entire sheath and the sutures due to interruptions. This means that if one suture is infected and loosens, it does not spread to the nearby suture, and because the tension is equally distributed amongst all the sutures, the rectus sheath is still in place. Interestingly, a vent is inadvertently created, allowing the collected from the affected site to drain through, helping other sutures stay intact. This benefits us, as easy closure can be planned later without planning a significant operation when the patient stabilizes. This has a substantial advantage over the continuous technique because if any of the bites of continuous suture give way, it weakens the entire fascial closure and makes early closures difficult . HoweverWhile the study provides valuable information on the clinical comparison between the far-near-near-far technique and conventional closure methods, several limitations should be acknowledged. Firstly, the study design is observational, which may introduce bias and confounding factors, and the relatively small sample size limits the findings' generalizability. A randomized controlled trial with a larger sample size would provide more substantial evidence for comparing the two closure techniques. Secondly, the study was conducted at a single center, which may limit the generalizability of the findings. Multi-center studies involving diverse populations would enhance the external validity of the results. Additionally, the surgeon's discretion in selecting the closure technique introduces potential bias, and a randomized controlled trial would mitigate this bias. Finally, the far-near-near-far group exhibited a shorter mean follow-up period than the continuous group, which may have implications for assessing long-term complications.In conclusion, this study contributes to the existing knowledge on rectus sheath closure techniques in emergency exploratory laparotomy. Our study provides initial insights into the clinical comparison between conventional rectus sheath closure and the far-near-near-far process in an emergency exploratory laparotomy. While we did not observe significant differences in postoperative abdominal wall complications between the two approaches, further research with larger sample sizes is warranted. The far-near-near-far technique may offer potential advantages in terms of tension distribution and is balanced in terms of operative time in contrast to popular belief. Still, additional studies are needed to confirm these findings. Surgeons should consider individual patient characteristics and optimize perioperative care to minimize complications in emergency abdominal surgeries. This study adds valuable insights to the surgical community, highlighting the need for comprehensive assessment when selecting abdominal closure techniques in emergency abdominal surgery.


Different techniques are being applied for automated vehicle counting from video footage, which is a significant subject of interest to many researchers. In this context, the You Only Look Once (YOLO) object detection model, which has been developed recently, has emerged as a promising tool. In terms of accuracy and flexible interval counting, the adequacy of existing research on employing the model for vehicle counting from video footage is unlikely sufficient. The present study endeavors to develop computer algorithms for automated traffic counting from pre-recorded videos using the YOLO model with flexible interval counting. The study involves the development of algorithms aimed at detecting, tracking, and counting vehicles from pre-recorded videos. The YOLO model was applied in TensorFlow API with the assistance of OpenCV. The developed algorithms implement the YOLO model for counting vehicles in two-way directions in an efficient way. The accuracy of the automated counting was evaluated compared to the manual counts, and was found to be about 90 percent. The accuracy comparison also shows that the error of automated counting consistently occurs due to undercounting from unsuitable videos. In addition, a benefit–cost (B/C) analysis shows that implementing the automated counting method returns 1.76 times the investment. The act of automated vehicle counting can be executed through a variety of approaches, including applying in situ technologies and computer algorithms. In situ technologies refer to counting traffic using detectors located along the roadside . There aThere are different traditional models for applying computer algorithms to count traffic. The basic procedure of all models is the application of the image processing technique . The comTo overcome these problems, Stauffer and Grimson  , Fast R-CNN, Single Shot Multi-Box Detector (SSD), and You Only Look Once (YOLO) . R-CNN aThe performance of YOLO was found to be superior to that of other neural-network-based object detectors in terms of accuracy and speed ,26. The Several studies have been conducted on in situ traffic counting technologies; for example, Patrizia Bellucci and Ernesto Cipriani  ; CountCam, and CountingCars  cameras. The video was collected for two successive days from 8 AM to 6 PM for each site. Later, these video data were processed using the developed automated traffic counting method to evaluate the model’s performance.The authors selected YOLO version 3 for this research. This version is significantly faster, accurate, and user-friendly . HoweverSince the proposed method does not involve any tiny object detection, the research results are also not affected by the other compared methods. For example, a recent (2023) version of the stochastic method proposed by Subhrasankha Dey et al.   [The YOLO weight file was originally developed using the C/C++ programming language. In order to apply the model in the Python preferable environment, the weight file was converted to TensorFlow architecture. The required packages for this conversion were TensorFlow 18.0 (a deep learning library), Numpy , OpenCV (an open-source computer vision library), TQDM (a progress bar library that provides useful routines for nested loops), and Python. The open-source YOLO weight file was downloaded from the official YOLO website (die.com) . After tdie.com) . The ‘pyFirst, this step defines the directory of the YOLO weight file, anchor definitions, YOLO model class definition, and the number of graphics processing units available for use. After that, the arguments were developed for the name and directory of the input pre-recorded video footage using the ‘parser’ task. All these developed arguments were executed using the function ‘__main__’, the fundamental function that manages the vehicle detection algorithms.The input video files must be processed before uploading to the program. All video files must be converted to the .mp4 format. If there are multiple video files, it is suggested that the video files be joined together because multiple video files cannot be uploaded simultaneously in the program. After processing the video file, it is uploaded in the section titled ‘input’ by dragging and dropping the file. The video file’s name is manually typed before the ‘.mp4’ title in the algorithm. When the program is executed, it checks for all the defined arguments and the input video file directory. If the program validates the directory and title of the video, it moves to the processing step.In this section, algorithms were developed to draw reference lines and detect vehicles in each video frame. The algorithms allow users to draw a reference line with simple mouse clicks when the very first video frame appears. The reference lines validate the program to make a successful vehicle count in the later steps. Though the detection of vehicles in each video frame was conducted by applying the converted YOLO weight file, a few algorithms were developed to execute the detection process, for example, the conversion of each image to a common size and providing threshold a value to identify potential detected objects. The total vehicle detection process is shown in When the input argument receives the name and the path of an input video file, the program calls the detector function (‘detect_start’) and passes the file information. Then the program calls another function (‘getFirstFrame’) to capture and display the first frame with the help of OpenCV, as shown in After that, the program calls the ‘detect_image’ function. The parameters of this function are a single video frame and reference lines. Under this function, a few more functions are defined, such as ‘get_right_line’ and ‘get_left_line’, which draw parallel lines on the right side and left side of the ‘mid_line’, respectively, as shown in The ‘detect_image’ function passes each video frame to the function ‘letter_box image’. This function converts each video frame to a common size (128 × 128 pixels) as required by the converted YOLO weight file in the TensorFlow API. Then, TensorFlow calls the converted YOLO weight file to detect objects in each frame ,46,47. ASince video footage consists of thousands of frames , an object must be tracked from one frame to another to determine its direction of movement . The maiIn this research, the tracker entitles each bounding box using a numeric value such as 1, 2, and 3. Rectangles are drawn around each tracked object using the function ‘draw.rectangle’, which was developed in the study. The rectangles are displayed on the computer screen, as shown in In this step, algorithms were developed to obtain the vehicle counts in a spreadsheet format with a flexible time interval and desired direction. This study requires counting vehicles in a flexible real-time interval, while YOLO’s counting speed differs from the real-time clock. To solve this problem, YOLO evaluates the number of frames in the desired time interval and the time to process a single frame. Then it estimates the total processing time of all frames in the desired time interval and considers that as the YOLO time interval. The algorithms are capable of providing vehicle counts in entry and exit directions.The algorithms determine the number and directions of counts based on the passing direction of the dotted center line through the mid-line and parallel line. For example, when the center line intersects the left parallel line first, the program confirms that the vehicle is coming from the left side. It can be assumed that the left side direction is the entry direction. Afterward, the program considers it a successful entry count when it intersects the mid-line. The developed ‘leftToright_counter’ and ‘rightToleft_counter’ functions count vehicles from the left and right sides, respectively.Counts are provided in a spreadsheet with the desired time interval. In addition, the counts are displayed on the computer screen. In order to allow YOLO to operate in a GPU environment at increased video processing speed, the model must be applied with OpenCV (Open-Source Computer Vision Library) before running the program. To accomplish this task, two open-source programs, cuDNN  and CUDA (NVIDIA’s programming language used to code the graphics card), are required. Both cuDNN and CUDA can be downloaded from the NVIDIA website  . CUDA muThe counting speed of the program depends on the configuration of the hardware platform. The following hardware platform was used in this study:Processor: Intel(R) Core (TM)i7-8750CPU @ 2.20 GHz 2.21 GHz;RAM: 2.7 GHz, 16.0 GB;GPU: NVIDIA GeForce GTX 1060, 6 GB.In this platform, it was found that the program takes about one hour forty minutes to one hour fifty minutes to process an hour of video. Using a high-configuration computer for processing videos is recommended to achieve increased video processing speed. A 2060 GPU with a video memory of more than 6 GB is recommended for timely processing.Cost of field data collection;Cost of conducting manual counts;Cost of energy consumption by computers.Lastly, a benefit–cost (B/C) analysis was conducted to evaluate the benefit of the automated counting method for practical implementation . The vidThe video data collection method and cost for the automated counts are the same as for the manual counts. Automated counting is conducted by computer algorithms using a hardware platform, and an individual is appointed to upload videos to the computer program. The individual is paid for the hours taken to upload videos in the program. In addition, before uploading videos, the individual pre-processes video footage, such as converting all videos to .mp4 format and joining videos. After uploading videos to the program, the computer is left for hours to process videos. From this research experience, it was found that it takes 15 min to process and upload 10 h of video to the program. This factor was applied to convert video hours to working hours. After that, the hourly pay rate was used to estimate the cost of automated counting.Cost of field data collection;Cost of conducting automated counts;Energy cost of configuring computer and setting model;Energy cost for running the model for video processing.There is considerable energy consumption in working with deep learning models. According to Neil C. Thompson et al.  (pp. 50–Equation (1) was used to estimate the benefit–cost (B/C) ratio of the automated counting method.k denotes a site;K denotes the total number of data collection sites;The pre-recorded videos were analyzed using the developed computer algorithms. Since the video was collected from 8 AM to 8 PM for each site for both day 1 and day 2, the total duration of video for an access point of a site was 20 h. The total number of access points for all 10 sites was 16. So, the total duration of video for all ten sites was 320 h. Therefore, in this automated counting method, a total of 320 h of video was processed. The entry and exit counts applying the automated counting method for the ten sites are shown in Automated counts were compared with manual counts to estimate the accuracy of the automated counting. The accuracy of a daily entry, daily exit, and total counts of individual sites for day 1 and day 2 are shown in t-test is a suitable statistical analysis to evaluate the differences between the observations of two variables. So, a two-tailed paired t-test was performed to evaluate the similarity between manual and automated counts. In this research, manual and automated counts were performed on ten sites. In this case, manual and automated counts are considered two variables, and sites are considered the same subject of interest.A two-tailed paired-A few assumptions were established for this test. It was assumed that independent variables  consist of two related groups , there are no significant outliers in the differences between manual and automated counts, and the distribution of differences between manual and automated counts shows an approximately normal distribution. The following hypotheses were considered:ai denotes automated total daily vehicle count at site i;Nmi = manual total daily vehicle counts at site i.Nt-test was conducted for two cases for day 1 and day 2, individually. In the first case, the t-test was conducted for the difference between manual and automated counts . In the second case, the test was conducted for the adjusted difference between manual and automated counts, where the adjustment was performed by the means of the difference . This second case was introduced to observe the effect of removing the undercounting bias in the results. The test results are shown in The paired In the case of the difference between manual and automated counts, i.e., t-test is that data shows a normal distribution pattern. Since the sample size of the paired t-test is small, a normality test was conducted to see whether the sample shows a normal distribution. In this case, the differences between manual and automated counts for day 1 and day 2 were individually analyzed. From the test, it was found that the data for the difference between manual and automated counts for day 1 and day 2 do not show a normal distribution. The reason for not showing normal distribution is random sampling and the small data size. Therefore, it may be a potential reason for not showing the similarity of manual and automated counting data in the paired t-test.However, one of the assumptions of a two-tailed paired Confidence intervals were estimated for the difference between paired manual and automated counts for day 1 and day 2, individually. The results are shown in The benefit–cost (B/C) analysis was conducted using Equation (1), as provided in the methodology section. The details quantity of the B/C analysis parameters are as follows.Pay rate of trained individuals = 10 USD per hour;Unit per of kWh energy  = USD 0.13 ;The hardware device used in this research is Lenovo legion Y7000P-1060 and the energy consumption is 183.96 Wh. ;K denotes the total number of data collection sites = 10 sites;  = energy consumption in 1 h * total hours  = 183.96 Wh. * 112 h = 20.60 kWh;  = energy consumption in 1 hr. * total hours  = 183.96 Wh. * 48 h = 8.83 kWh;  = energy consumption in 1 h * total hours  = 183.96 Wh. * 480 h = 88.30 kWh; = 16 * USD 10 = USD 160; = kWh * unit price = 20.60 * USD 0.13 = USD 2.68;  = kWh * unit price = 8.83 * USD 0.13 = USD 1.15;  = kWh * unit price = 88.30 * USD 0.13 = USD 11.18.Using Equation (1) and the factors described above, the benefit–cost (B/C) analysis is conducted as follows:The estimated B/C ratio is 1.54, which implies that the proposed automated counting method has a 54 percent greater benefit then the traditional manual counts.The accuracy of different automated counting methods, including this study, is shown in Pneumatic road tube counting: Patrick McGowen and Michael Sanderson  (p. 1) cPiezoelectric sensor: When a vehicle passes over a piezoelectric sensor, mechanical energy passes to the sensor, thereby converting mechanical energy to electrical energy, which is analyzed for vehicle counting. The accuracy of traffic counts applying the piezoelectric sensor is reportedly 99 percent  (pp. 175Inductive loops: According to Chen-Fu Liao  (p. 1), Computer algorithms: Mattias Gustafsson and Sebastian Hjelm  , traffic congestion monitoring, parking management, traffic safety analysis, and transit planning. It was found from the study that the proposed model has some excellent advantages, such as providing traffic counts with 90 percent accuracy in a consistent manner; providing directional vehicle counts with flexible time intervals as required by many engineers, planners, and policymakers; and being economical, especially for small transportation projects. On the other hand, the proposed model has some drawbacks, such as the program not being efficient enough when vehicles pass at high speed, and the program compromises accuracy when the video quality is poor, especially when recorded during rain, low light, evening, and cloudy weather. In addition, the wrong camera angle and vehicles overlapping during recording may reduce the model’s accuracy. To overcome the drawbacks, it is recommended to use high-resolution cameras to obtain good-quality videos, properly install the cameras to avoid oscillation due to wind, and clean the camera lens before every installation. Moreover, it is recommended to use high-resolution computers to increase the speed of counting and save energy. A benefit–cost analysis was conducted considering different factors, including the energy consumption of the system. From the benefit–cost (B/C) analysis, the B/C was found to be greater than or equal to 1.54, which means the automated counting method has 54 percent greater benefit than the traditional manual counting method. In other words, it can be implied that the model can be suitable for the practical application of vehicle counting, especially for projects that cannot afford in situ counting technologies.This study presents an efficient method of automated vehicle counting from pre-recorded video by applying a pre-trained YOLO object detector in TensorFlow API. The accuracy of automated counting was found to be about 90 percent, with automated counting consistently being undercounted. This accuracy is very significant compared to the other automated counting method. The average processing speed for an hour of video was found to be about one hour and thirty minutes. The advantage of the model is that it can provide 90 percent accuracy of directional counts with a flexible interval time in a consistent manner. On the other hand, the drawback of the model is that it compromises accuracy when the vehicles pass at high speed and overlap, the video frame is narrow, and the video quality is poor. From the benefit–cost (B/C) analysis, the B/C was found to be greater than or equal to 1.54, which implies that the automated method improves the benefit by 54% compared to the traditional manual method. Therefore, the developed algorithm might be an excellent replacement for the other expensive automated counting methods.


Higher serum Mg2+ levels, increased dietary Mg2+ in-take, and Mg2+ supplementation are each associated with lower blood pressure, suggesting that Mg2+-deficiency contributes to the pathogenesis of hypertension. Antigen-presenting cells, such as monocytes and dendritic cells, are well-known to be involved in the pathogenesis of hypertension. In these cells, processes implicated as necessary for increased blood pressure include activation of the NLRP3 inflammasome, IL-1β production, and oxidative modification of fatty acids such as arachidonic acid, forming isolevuglandins (IsoLGs). We hypothesized that increased blood pressure in response to dietary Mg2+-depletion leads to increased NLRP3, IL-1β, and IsoLG production in antigen presenting cells. We found that a Mg2+-depleted diet (0.01% Mg2+ diet) increased blood pressure in mice compared to mice fed a 0.08% Mg2+ diet. Mg2+-depleted mice did not exhibit an increase in total body fluid, as measured by quantitative magnetic resonance. Plasma IL-1β concentrations were increased (0.13 ± 0.02 pg/mL vs. 0.04 ± 0.02 pg/mL). Using flow cytometry, we observed increased NLRP3 and IL-1β expression in antigen-presenting cells from spleen, kidney, and aorta. We also observed increased IsoLG production in antigen-presenting cells from these organs. Primary culture of CD11c+ dendritic cells confirmed that low extracellular Mg2+ exerts a direct effect on these cells, stimulating IL-1β and IL-18 production. The present findings show that NLRP3 inflammasome activation and IsoLG-adduct formation are stimulated when dietary Mg2+ is depleted. Interventions and increased dietary Mg2+ consumption may prove beneficial in decreasing the prevalence of hypertension and cardiovascular disease.Nearly 30% of adults consume less than the estimated average daily requirement of magnesium (Mg Decades of work demonstrate that systemic inflammatory activity increases blood pressure . More reSeveral lines of evidence suggest the importance of the NLRP3 inflammasome in the pathophysiology of hypertension. In Dahl salt-sensitive rats, in addition to experiencing increased blood pressure, a high salt diet increases expression of NLRP3, ASC, and caspase-1 . In cont+ T cells  can promote hypertension through the production of isolevuglandins (IsoLGs) . These γ T cells . CD8+ T  in mice .2+ deficiency is associated with activation of the NLRP3 inflammasome and increased production of IsoLGs. We confirmed that dietary Mg2+ depletion in a mouse model provokes hypomagnesemia and increases blood pressure. We examined body composition of Mg2+-depleted mice to determine whether an increase in body fluid could contribute to this increase in blood pressure. We examined antigen-presenting cells in spleen, kidney, and aorta, discovering that Mg2+ depletion increased NLRP3 and IL-1β in these cells in vivo. Furthermore, we found that Mg2+ depletion increased in vivo formation of IsoLG-adducts. To determine whether reduced extracellular Mg2+ could provoke these effects directly, we examined CD11c+ dendritic cells in primary culture and found that reduced extracellular Mg2+ also increases IL-1β production. These findings suggest that systemic inflammation occurring in response to dietary Mg2+ insufficiency may contribute to the pathogenesis of hypertension.Dietary magnesium also influences blood pressure. Multiple population-based cross-sectional studies and clinical trials observed a high prevalence of magnesium deficiency with 10%–30% of individuals having serum magnesium concentrations below the standard cutoff (<0.80 mmol/L) . Dietaryin vivo experiments were performed in mice in the SV129 (129S2/SvPasCrl) background, obtained from Charles River Laboratories. Due to the observation that female mice are relatively resistant to inflammation-associated hypertension, only male mice were used in this study (2+-deficient diet for mice was purchased from Envigo (Teklad TD.93106) with starting Mg2+ content of 0.0015%–0.003% and Na+ content of 0.15%. To this, Mg2+ oxide (Sigma) was added back to achieve final Mg2+ content of either 0.01%, 0.02%, or 0.08%, and in one group NaCl was added to a final Na+ content of 4% as noted in results section and figure legends. Powdered diets were provided to mice in cages of three to five animals in tip-proof feeding jars for powdered diets . Food consumption was assessed by weighing jars daily and dividing decrease in weight by the number of mice in the cage. As a positive control for NLRP3 inflammasome activation and IsoLG production in antigen-presenting cells as previously shown , as per manufacturer directions. Circulating cytokines were measured in plasma using a V-Plex®and V-Plex®Plus Mouse Proinflammatory Panel Kit .Plasma MgBody water content was measured in non-anesthetized mice using a quantitative magnetic resonance body composition analyzer (EchoMRI) as previously described .+/MerTK−/I-A/I-E+/CD11c+. Monocytes were identified as being CD45+/MerTK−/I-A/I-E+/CD11c+/CD115+. Representative flow cytometry plots were selected to best represent the gating strategy.Harvested spleens were placed in spleen dissociation buffer and dissociated using a 3 mL syringe plunger. Harvested kidneys were placed in kidney digestion buffer  and DNAse I  and thoracic aortas with surrounding perivascular fat were placed in aorta digestion buffer , Collagenase B , and DNAse I  and placed in a rotating 37°C incubator for 30 min. Tissue homogenates were then passed through a 40 μm strainer and spun down at 400 × g for 10 min at 4°C. Kidney single cell suspensions underwent a Ficoll gradient centrifugation to isolate leukocytes, whereby kidney pellets were resuspended in 36% Ficoll in PBS and gently placed on top of 72% Ficoll in PBS in 15 mL tubes and centrifuged at 2,400 rpm for 15 min with decreased deceleration to prevent mixing of the density layers. Leukocytes are collected from the top density layer, diluted with PBS, and centrifuged. Single-cell suspensions were pretreated with FcR blocking reagent  and subsequently stained with LIVE/DEAD™ Fixable Violet Dead Cell Stain Kit (ThermoFisher) to determine cell viability. For surface staining the following fluorophore-conjugated antibodies were used (1 μg/100 μL): Brilliant Violet 510 anti-CD45 , PE-Cy7 anti-MerTK , PerCP-Cy5.5 anti-I-A/I-E , PE-Fluor 610 anti-CD11c , APC-Cy7 anti-CD115 . Cells were then fixed and permeabilized for incubation of intracellular markers. For intracellular staining, we used PE anti-IL1β , D11 ScFv antibody to detect IsoLGs which was conjugated to Alexa Fluor 488 microscale labeling Kit , and anti-NLRP3  which was conjugated to Alexa Fluor 647 Antibody Labeling Kit . Samples were run on a Cytek Aurora system and analyzed using FlowJo Software . Gates were set live singlets and subsequent gates were based on flow minus one (FMO) controls for each fluorophore. Results are expressed as percent positive per either dendritic cell or monocyte. Dendritic cells were identified as being CD452+ medium for cell culture was prepared using Gibco™ Mg2+-free RPMI 1640 (Life Technologies Corp.) with L-glutamine, supplemented with 1% penicillin-streptomycin, 15 mM HEPEs, 1 mM sodium pyruvate, 0.05 mM β-mercaptoethanol, and 10% fetal bovine serum. Control Mg2+-medium was supplemented with MgCl2. Final mineral concentrations were measured using inductively coupled mass-spectrometry (ICP-MS), which showed concentrations of 0.12 mM in the low Mg2+ solution and 0.90 mM magnesium in control Mg2+ solution, which is within the normal range of blood magnesium of 0.85–1.10 mM (2+ medium). Dendritic cells were isolated from a single-cell suspension of splenocytes by magnetic labelling and positive selection of CD11c+ cells using Miltenyi Biotec Isolation Kit  according to the manufacturer’s protocol using LS columns. Cells were plated in 24-well plates at a density of 1 × 106/mL in either low or control Mg2+ medium for a variable period, as described in Results section. In some experiments, cells were pre-treated for 20 min with or without YVAD  or MCC950 . IL-1β production was measured using a commercially available ELISA kit (Abcam) according to manufacturer instructions.Low Mg–1.10 mM . ConcentN value. All results are presented as mean ± standard error of the mean. Normality of distribution was assessed using the Shapiro-Wilk test. Testing for outliers was performed using the ROUT method (Q = 1%). Comparisons of two groups were performed using Student’s t-tests for parametric data or Mann-Whitney test for non-parametric data. A repeated-measures analysis of variance (ANOVA) followed by a Bonferonni’s multiple comparison test were used to compare more than two groups. A comparison of more than two non-parametric groups having an N less than 6 where normality cannot be properly tested , we used a two-way ANOVA with a Dunnett’s post hoc multiple comparison. For all analyses, a two-tailed p-values < 0.05 was used to reject the null hypothesis. Analyses were performed using Graphpad Prism 8.4.2.In all data sets, one mouse represents 1 d i.e.,: –6 a Krus2+ deficiency increases blood pressure in a SV129 background mouse model. This strain of mice was chosen because of its sensitivity to hypertensive stimuli relative to other strains of mice, such as C57B/6J (2+ diet (n = 7), a magnesium deficient diet, developed higher blood pressures  than in those receiving 0.08% Mg2+ diet (n = 7)  and 0.08% Mg2+  diet groups developed significantly higher systolic blood pressure at the end of the experiment compared to baseline  compared to mice on the 0.08% Mg2+diet , experienced a statistically significant dip in weight after the first week on the powdered diet compared to the 0.01% Mg2+ diet group (27.9 ± 0.5 g) (Pinteraction = 0.47) (2+ diet group prompted termination of the experiment and euthanization of the mice (We confirmed that dietary Mg C57B/6J . Blood p (n = 7) . Howeverbaseline . Mice weMg2+diet . Mouse w± 0.5 g) . Food co = 0.47) . Survivathe mice .2+ on body composition. A milder Mg2+-deficient diet  was employed to avoid the mortality observed in 2+ (n = 7) diet induced a significant reduction  in plasma Mg2+ as compared to the 0.08% Mg2+ diet (n = 7), however it should be noted that the difference between 0.08% Mg2+ diet and the 0.02% Mg, diet was reduced compared to the difference observed in mice from Ptime = 0.02; 2+ for one group and 7 days of 0.02% Mg2+ for the other group) total body water decreased in both groups by about 10% . Thus, we found no evidence that dietary Mg2+ depletion altered total body fluid.To evaluate whether the observed increase in blood pressure was associated with an increase in total body fluid, we examined the influence of dietary Mg2+ deficiency induced systemic changes in inflammatory cytokines. Cytokines were measured in plasma of mice euthanized at the end of the experiment in 2+ or 0.02% Mg2+ diet. We found a significant increase in IL (interleukin)-1β in mice on the 0.02% Mg2+  diet  diet . Point e+ leukocytes decreased in response to the 0.01% Mg2+ diet with no changes in either dendritic cell or monocyte numbers  but not dendritic cells increased significantly  and dendritic cells .Plasma cytokine levels are a relatively insensitive indicator of systemic inflammation. We therefore looked more directly at expression of IL-1β in splenic antigen-presenting cells (monocytes and dendritic cells). Single cell suspensions prepared from spleens of mice euthanized at the end of the experiment in  numbers . Additio numbers . We founficantly . The per2 , but not monocytes, in mice subjected to a high salt diet  but not monocytes, suggesting that Mg2+ depletion is a potent stimulus of this pro-hypertensive inflammatory pathway.Additionally, we examined isolevuglandins (IsoLGs) in splenic antigen-presenting cells. These γ-ketoaldehydes arise through oxidation of fatty acids and their phospholipid esters, in particular prostaglandin H2 . In anim2 . We obsealt diet . Dietary2+, 8.5% ± 2.0% versus 0.08% Mg2+, 0.6% ± 0.5%; p = 0.002), but not dendritic cells or total CD45+ leukocytes, in mice fed a 0.01% Mg2+ diet  as well as the mice fed a high salt diet  when compared with the 0.08% Mg2+ diet group , as was the percentage that was positive for NLRP3 ,  and the high salt diet  increased the percent IsoLG positive kidney dendritic cells compared to the 0.08% Mg2+ diet , . As withg2+ diet . These r2+ diet significantly increased the number of monocytes per aorta  but not dendritic cells nor total CD45+ leukocytes  and dendritic cells , even though the high salt diet only increased NLRP3 percent positivity in dendritic cells , but not monocytes  and dendritic cells  compared to the 0.08% Mg2+ diet  or control (0.90 mM) Mg2+. By 6 h, both IL-1β levels , a caspase-1 inhibitor (5 μg/mL YVAD) or an inhibitor of NLRP3  . Similarly, we observed an increased release of IL-18, another NLRP3 inflammasome activation marker . Both YVAD and MCC950 significantly attenuated this increase in IL-1β  . Eighteeβ n = 5,  concentrr n = 6, , in the 2+ depletion increases blood pressure in a mouse model. This increase in blood pressure is achieved with no significant difference in total body fluid. Dietary Mg2+ depletion also stimulated systemic inflammation, as seen by an increase in circulating IL-1β levels. Mg2+ depletion increased NLRP3 expression in monocytes from spleen and aorta and in dendritic cells from spleen, kidney, and aorta. Mg2+ depletion also induced IsoLG production in monocytes from spleen and aorta and in dendritic cells from spleen, kidney, and aorta (2+-depletion on these markers of inflammation were generally comparable to, or greater than, the effects of a high salt diet. Finally, Mg2+-depletion in dendritic cells in culture induced IL-1β production, confirming a direct stimulatory effect.The present study demonstrates that moderate dietary Mgnd aorta . The eff2+, in contemporary fruits and vegetables has declined by 80% or more compared to Mg2+ in produce grown in the early 20th century (2+ is also reduced by modern processing methods (2+ requirement (2+ depletion (2+ wasting, promoting hypomagnesemia (In humans, systemic magnesium depletion is common. Content of essential minerals, including Mg century . Food Mg methods . As suchuirement . Moreoveepletion . Thiazidgnesemia . Hydrochgnesemia . Thus, M2+-depletion promotes hypertension. In Wistar rats given a Mg2+-deficient diet, blood pressure increased (2+ supplementation attenuated increases in blood pressure in both angiotensin II and high salt diet rat models of hypertension (2+-depletion. An early study in Wistar rats showed decreased blood pressure (Mgncreased . Dietaryrtension . Howeverpressure . Animalspressure . In that2+ protects against increased blood pressure. Lower serum Mg2+ was found to be associated with increased odds of hypertension (2+ in-take was associated with decreased risk of hypertension in a meta-analysis of ten cohort studies (2+ (2+ balance influences blood pressure in humans.Human studies suggest that dietary Mgrtension . Dietary studies . Meta-andies (2+ . This efdies (2+ . These s2+ depletion could increase blood pressure through numerous mechanisms. In our study, although a low Mg2+ diet increased blood pressure, it did not increase total body fluid. The decreased difference in plasma Mg2+ level observed in mice examined for body composition, as compared to mice examined for blood pressure, represents a caveat in the interpretation of these findings. It is possible that a difference in body water may have been observed if experimental and control groups had a bigger difference in plasma Mg2+ or if body water had been measured at more frequent time intervals. Nevertheless, the data do not provide evidence for a difference in body water associated with hypomagnesemia.Systemic Mg2+ levels were associated with decreased eGFR (2+ levels associated with higher risk for end stage renal disease (2+ diet exhibited elevated systolic blood pressure and increased NLRP3 inflammasome activation in kidney dendritic cells, which may contribute to intrarenal inflammation associated with magnesium deficiency-induced hypertension.Hypertension and kidney disease are closely linked whereby hypertension can also cause as well as contribute to kidney disease progression . Incidensed eGFR . Additio disease . In our 2+ diet. Increased systemic Mg2+ has been shown to increase arterial elasticity (2+ mediates vasorelaxation through its action as a calcium channel antagonist, regulating calcium influx (2+ depletion in the present study may be the result of increased inflammatory and oxidative stress, as shown by NLRP3 inflammasome activation and IsoLG-adduct formation, respectively, of antigen presenting cells in the aorta. Future studies examining systemic vascular resistance and cardiac output may shed further insight on the mechanisms of how Mg2+ deficiency induces increases in blood pressure.Differences in arterial elasticity may contribute to the observed increase in blood pressure in mice on a low Mgasticity . Arteriaasticity . Mg2+ mem influx . Hypomagm influx . Given t2+ depletion. Both myeloid-derived NLRP3 inflammasome and IsoLGs contribute to the pathogenesis of hypertension (2+ concentration activates IL-1β production in cell culture demonstrates a direct stimulatory effect of Mg2+-depletion on dendritic cell activity. NF-κB is a transcription factor responsible for upregulation of NLRP3 inflammasome components. A study using neonatal monocytes found that magnesium sulfate supplementation inhibits NF-κB activity (2+ activates these cells will require further study.Magnesium plays a role in multiple cellular functions, and its deficiency has a significant impact on inflammatory processes. Limiting dietary magnesium by just 10% induces leukocyte migration, tissue infiltration, and activation to release inflammatory cytokines and free radicals . During rtension . This stactivity . However2+ depletion activates the NLRP3 inflammasome and stimulates production of IsoLGs, both of which promote hypertension. Given the prevalence of hypertension and of dietary Mg2+ depletion in human populations, the findings suggest that dietary Mg2+ consumption may represent an important, modifiable risk factor for hypertension and cardiovascular disease.This study represents the first study showing that dietary Mg