In this article, we report results from a nationwide survey on pandemic-related health behavior in Russia. A total of 2,771 respondents aged 18 to 82 were interviewed between January 21 and March 3, 2021. The survey included questions on perceived vulnerability to coronavirus, prevention-related health behavior, readiness for vaccination, and general awareness about COVID-19. Descriptive data showed that 21.2% of respondents reported high vulnerability to the coronavirus, and 25% expressed fear. Moreover, 38.7% of the surveyed individuals reported low trust in vaccination efficacy, and 57.5% were unwilling to take a vaccine, which was much higher than the official data. Based on the evidence obtained, four types of health behavior during the pandemic were constructed. Rational (29.3%) and denying (28.6%) behaviors prevailed in men, while women were found to more likely behave with a vaccine-hesitant demeanor (35.7%). Educational background affected the proportion of respondents with the denying type of health behavior, who were also of younger age. The rational behavioral type was found to be more common among respondents aged above 50 years and prevailed as well among individuals with university degrees. The middle-aged population of Russia was highly compliant with prevention-related health practices; however, vaccine hesitancy was also high among them. Furthermore, health behaviors varied significantly across the Federal Districts of Russia. We are convinced that our results contribute to existing public health practices and may help improve communication campaigns to cause positive health behaviors. The current coronavirus variant of EG.5 was evaluated by the World Health Organization as having low public health risk at the global level  aged 16 to 82 (mean age 25.6 ± 10.8 years), who were residents of the Central Federal District of Russia (40%), Northwestern Federal District (10.4%), Volga Federal District (27.3%), Southern Federal District (10.6%), and Siberian Federal District (6.6%). In all, 5.1% of the respondents preferred not to disclose their place of residence, and 729 respondents (26.3%) reported a history of COVID-19 disease. Most of them reported a mild form of the disease (87.8%), while 12.2% indicated a history of severe COVID-19. Detailed participants’ characteristics are presented in Participants’ degree of compliance with prevention-related behavioral practices was assessed via Question 8. The answer options consisted of a five-point Likert scale, ranging from “Never” to “Always.” “Never” and “Rarely” responses to any of the Q8 subquestions were encoded as low compliance with preventive practices. Attitudes toward vaccination were investigated via Question 9 and Question 10 based on a three-point Likert scale with answer options ranging from “Disagree” to “Agree.” The response “Disagree” to Q9 or Q10 was weighed as low trust in the COVID-19 vaccine. To allocate complex behavioral types during the pandemic, the data of low vs. high compliance with preventive practices and low vs. high trust to vaccination were aggregated and analyzed.χ2) statistic were applied.R software was used to process the data. Measures of frequency and chi-square . All the respondents signed an electronic informed consent form.3.3.1.To investigate perceived vulnerability to coronavirus infection, preventive behavior, COVID-19 vaccination attitude, and coronavirus awareness during the second wave of the pandemic in Russia, data on the survey participants’ responses were analyzed. At first, we found that 25% of all the respondents were afraid of catching COVID-19, having answered “A lot” (5.7%) and “Quite afraid” (19.3%) to the question “Are you afraid of catching COVID-19?.” A total of 21.2% of the respondents perceived themselves as vulnerable to the infection by agreeing with the answers “Extremely” (4.2%) and “Rather highly” (17%) to the question “How vulnerable are you to COVID-19?.” In addition, 29.8% of the survey participants reported a relatively high probability of future infection, having answered “Very high” (11.1%) and “Quite high” (18.7%) to the question “What are your chances of catching COVID-19?” Detailed response data are presented in Regarding preventive practices, particularly COVID-19 prevention, the majority of the respondents always or at least often followed existing recommendations. They reported washing hands with soap after visiting public places , wearing a face mask in public places (89.8%), avoiding touching their eyes, nose, and mouth with unwashed hands (62.3%), using hand sanitizers (58.6%), and physical distancing in public places (55.6%) .Attitudes toward the COVID-19 vaccine and readiness to vaccinate were of particular interest in this January–March 2021 survey. The survey statement “I will agree to take the COVID-19 vaccine” received 57.7% negative, 19.8% positive, and 22.5% “Do not know” responses. In addition, 38.7% of the respondents disagreed that “a vaccine can help control the spread of COVID-19,” 31.6% said they did not know, and 29.7% of the respondents agreed. The statement “COVID-19 vaccination should be mandatory for some groups” was supported by 26.1% of the respondents, while 49.9% disagreed. Detailed response data are presented in Data on coronavirus awareness showed that only 24.6% of the respondents believed coronavirus was of a natural origin. Meanwhile, 9.5% believed that “COVID-19 was invented in a laboratory with a purpose,” and 49.1% of the survey participants considered a laboratory origin of COVID-19 to be possible or found it difficult to answer .Based on the data obtained and considering the dimensions of (1) compliance with prevention-related behavioral practices, low vs. high, (2) trust in a COVID-19 vaccine, including readiness to vaccinate, low vs. high, four behavioral types were constructed for further analysis.Type 1 “Rational”: A total of 753 respondents (27.2%), who reported high compliance with prevention-related behavioral practices and high trust in a COVID-19 vaccine;Type 2 “Vaccine hesitant”: A total of 892 respondents (32.2%), who reported high compliance with prevention-related behavioral practices but low trust in a COVID-19 vaccine;Type 3 “Denying”: A total of 762 respondents (27.5%), who reported low compliance with preventive practices and low trust in a COVID-19 vaccine;Type 4 “Inconsistent”: A total of 364 respondents (13.1%), who reported low compliance with preventive practices but high trust in a COVID-19 vaccine.On the whole, almost one-third (27.2%) of the surveyed individuals behaved in accordance with recommendations on COVID-19 prevention and were ready to take a vaccine, and another third (27.5%) did not comply with recommendations and showed no trust in preventive practices and vaccination. Another third of the respondents (32.2%) expressed vaccine hesitancy, but they maintained preventive behavior, while 13.1% of the surveyed individuals reported trust in a COVID-19 vaccine but not in COVID-19 preventive practices. Next, we analyzed the demographic differentiations of the behavioral types.3.2.χ2  = 41.466, p < 0.001. The data on distribution are provided in The respondents’ behavioral-type distribution by sex showed that the major part of male respondents comprised “Rational” and “Denying” behavioral types , while the female respondents were found to keep mostly “Vaccine-hesitant” behavior (35.7%), n = 921), 20–29 , 30–39 (n = 278), 40–49 (n = 221), 50–59 (n = 86), and 60 years and above (n = 51). The results showed that age proportions significantly differed by type, χ2  = 96.008, p < 0.001. A weighty percentage of respondents under the age of 30 showed “Denying” and “Vaccine-hesitant” behavior . They all reported low trust in a COVID-19 vaccine, and the younger they were, the lower compliance was with prevention-related behavioral practices. The age groups of 30–39 and 40–49 showed congruent results in prevention – the vast majority of these middle-aged individuals were highly compliant with preventive recommendations. However, they either reported trust in vaccines  or had low confidence in vaccination efficacy . At the same time, a significant part of the respondents of older ages (50–59 and 60 years and above) were found to have the “Rational” behavioral type. Most of them  reported compliance with preventive practices and trust in vaccination. To investigate the age-specific distribution of the behavioral types, we grouped the respondents by their age. We considered age groups of under 20 years  = 45.917, p < 0.001, and showed that most of the respondents with incomplete secondary education (40%) comprised the “Denying” behavioral type. At the same time, individuals with secondary and vocational secondary education had “Denying” (30.6 and 33.3%) and “Vaccine-hesitant” (29.4 and 33.9%) health behaviors. The hesitant type was also registered in most of the respondents with incomplete higher education (33.9%), while “Rational” health behavior was found to prevail among individuals with higher education (33%). Detailed distribution data are available in Since education is widely considered as a factor that influences perceptions of ongoing events and corresponding behavior, including social and health behavior , we analχ2  = 47.635, p < 0.001 (see Most respondents who believed in COVID-19 conspiracy theories showed “Denying” behavior (41.5%), whereas individuals with no reported conspiracy beliefs were inclined to “Vaccine-hesitant” (32%) and “Rational” (28.7%) behavioral types, .001 see .3.3.χ2  = 69.26, p < 0.001  showed that only 32% of the surveyed participants expressed a negative attitude toward vaccination, only 4% did not want to take a vaccine, and 37% of respondents declared that they have already been vaccinated or found it difficult to answer the question  in our study disagreed to take a COVID-19 vaccine, and 38.7% also disagreed that a vaccine can help control the spread of the coronavirus. Interestingly, nine months later, the available data from November 2021 reported by the Russian Public Opinion Research Center via a telephone-based survey methodology  showed that more than 31% agreed that coronavirus was intentionally created and spread . This corresponds to the well-described gender differences in behavior  and the The highest rate of COVID-19 conspiracy beliefs (41.5%) was registered among the respondents with “Denying” health behavior, which corresponds, to a certain extent, to the opinions and behaviors interrelation model , 24.Along with that, educational background was found to affect the proportions of respondents with “Rational” and “Denying” behavioral types by doubling the rate of the former from 17.5% among respondents with incomplete secondary education to 33% among individuals with university degrees and by decreasing the rate of the latter from 40 to 22%. “Denying” individuals were also younger (less than 30 years), while “Rational” were older (50 years and above), as older age was and still is a pandemic-related risk factor for heath. The middle-aged population of Russia (30–39 and 40–49 years of age) was highly compliant with prevention-related health practices; however, there were also high rates of vaccine-hesitant behavior among them. As the middle-aged population is most economically active, they should be considered for special targeting when planning a prevention campaign and vaccination promotion.Despite the significant differences in health behaviors that we found across the Federal Districts of Russia, this study was not aimed to comprehensively address regional and cross-regional tendencies. Given the great variability of environmental factors, social capital, cultural health beliefs, and pandemic-related public health policies among the regional units within the Federal Districts, further research is needed to understand the dimensions of health behavior at a regional level.As the national healthcare agenda is focused on pandemic-related somatic burden , existin5.Our findings contribute to the existing knowledge of health behavior and its determinants. Due to vaccine distrust among the Russian population and the country’s slower vaccination campaign compared to most other European countries during the pandemic, the results we have reported may improve disease prevention and advance communication campaigns to cause positive health behaviors.peshkovskaya@gmail.com.The datasets presented in this article are not readily available because of ethical restrictions. Requests to access the datasets should be directed to AP, The studies involving humans were approved by the Ethics Council of Tomsk State University. The studies were conducted in accordance with the local legislation and institutional requirements. The participants provided their written informed consent to participate in this study.AP: Conceptualization, Funding acquisition, Methodology, Validation, Project administration, Resources, Supervision, Writing – original draft, Writing – review & editing. SG: Data curation, Formal analysis, Investigation, Methodology, Writing – original draft.The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This work was supported by Grant no. 075-15-2022-1152 .The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Medical imaging is an important tool for accurate medical diagnosis, while state-of-the-art image reconstruction algorithms raise critical challenges in massive data processing for high-speed and high-quality imaging. Here, we present a memristive image reconstructor (MIR) to greatly accelerate image reconstruction with discrete Fourier transformation (DFT) by computing-in-memory (CIM) with memristor arrays. A high-accuracy quasi-analogue mapping (QAM) method and generic complex matrix transfer (CMT) scheme was proposed to improve the mapping precision and transfer efficiency, respectively. High-fidelity magnetic resonance imaging (MRI) and computed tomography (CT) image reconstructions were demonstrated, achieving software-equivalent qualities and DICE scores after segmentation with nnU-Net algorithm. Remarkably, our MIR exhibited 153× and 79× improvements in energy efficiency and normalized image reconstruction speed, respectively, compared to graphics processing unit (GPU). This work demonstrates MIR as a promising high-fidelity image reconstruction platform for future medical diagnosis, and also largely extends the application of memristor-based CIM beyond artificial neural networks. Image reconstruction algorithms raise critical challenges in massive data processing for medical diagnosis. Here, the authors propose a solution to significantly accelerate medical image reconstruction on memristor arrays, showing 79× faster speed and 153× higher energy efficiency than state-of-the-art graphics processing unit. Magnetic resonance imaging (MRI) and computed tomography (CT) are two representative medical imaging technologies8. Although adopting different imaging principles, their imaging processes can be roughly divided into two fundamental steps: signal acquisition and image reconstruction   filter34 and discrete Fourier transformation (DFT)36, which has the potential to significantly accelerate medical image reconstruction speed and reduce energy consumption. In both applications, the most computationally intensive computations are vector-matrix multiplication (VMM); however, their actual implementations on memristor arrays are quite different in two aspects. Firstly, the entries in the matrix, i.e., synaptic weights, for ANNs are trainable, so that the effect of non-ideal device characteristics of memristors  may be accommodated by training31. In comparison, the entries in the matrix for signal processing algorithms, e.g., DFT coefficients, are pre-calculated without any training, so they are more susceptible to those non-ideal device characteristics, which could lead to large errors in the output. Secondly, while the entries in the matrix for ANNs are usually real numbers, the matrices used in signal processing usually have both real and imaginary parts, and directly mapping them onto different memristor arrays (as proposed in literature36) could result in large overhead in both energy consumption and area. Therefore, a new scheme to efficiently implement signal processing algorithms such as DFT on memristor arrays needs to be developed.Fortunately, computing-in-memory (CIM) technology based on emerging nonvolatile memories, such as memristors, can provide an alternative solution for medical image reconstruction with ultrahigh efficiency to break the von Neumann bottleneck. In this paradigm, computations are carried out at the place where data are stored through physical laws, largely reducing the energy-intensive data movementNs) Fig. , showing38, to segment and extract organs from MIR and software-reconstructed images, achieving similar DICE scores around 0.98. Besides, MIR shows 153× advantage in energy efficiency and 79× advantage in normalized image reconstruction speed for CT reconstruction task, compared to CMOS systems, indicating its great potential in low-power and high-speed portable medical imaging system for future medical scenarios.In this work, we propose and demonstrate a memristive image reconstructor (MIR), whose core is a memristive DFT Fig. . To effi31, which consists of eight 2K-cell memristor arrays. Figure x/HfO2/TiN, where HfO2 is the resistive switching layer and TaOx serves as the thermal enhanced layer to improve the analog switching characteristics  complexity for conventional DFT implementation or O(NlogN) complexity for fast Fourier transformation (FFT).To demonstrate the functionality of the MIR system, we first implement a 64-point one-dimensional (1D) complex DFT on the memristor arrays with 16 K memristors. DFT is a widely used time/spatial-frequency analysis technology and plays a key role in many applications including denoising, imaging and communication. As shown in Fig. N × N would be needed to map the matrix entries onto memristor conductance for the implementation of a N-point DFT36, as shown in Fig. As the DFT coefficients matrix has both real and imaginary parts, conventionally, four independent memristor arrays of the same size 41  strategy to eliminate the quantization errorFigure To evaluate the overall performance of our MIR in data-intensive medical imaging applications, we further implement an end-to-end MRI image reconstruction and segmentation task. Figure 42, which contains 30 three-dimensional (3D) MRI images covering the entire human heart. As the ground truth, the left atrium was segmented by an expert using an automated tool followed by manual corrections. Figure Here we use the heart dataset provided by King’s College London43 along with intermediate results.In the above demonstration of MRI image reconstruction, only one step of 2D DFT is carried out. In more advanced medical imaging algorithms, more than one steps of transformation are needed, where the impact of cumulative errors could degrade the image quality. As shown in Supplementary Fig. 44. In this dataset, the spleen was segmented semi-automatically and then were manually modified by an expert abdominal radiologist, serving as the ground truth. Figure Here, we use the spleen dataset from Memorial Sloan Kettering Cancer Center, including 61 portal venous phase CT scansIn conclusion, we have proposed and experimentally demonstrated a memristive medical image reconstructor, MIR, which shows excellent performance in energy efficiency and computing speed and is also highly robust to the non-ideal device characteristics of memristors. With the developed QAM strategy and CMT method, MIR could efficiently implement high-accuracy 1D/2D DFT and IDFT. In the MRI and CT image reconstruction tasks, compared with Nvidia Tesla V100 GPU, MIR has achieved a software-equivalent image reconstruction quality in terms of PSNR and DICE score, and meanwhile exhibited 112× and 153× improvements in energy efficiency, and 36× and 79× improvements in the normalized image reconstruction speed, respectively. Such computational advantages could be further improved by using larger memristor arrays and further optimizing the memristor analog switching characteristics. This work suggests that our MIR system could provide a high-speed and energy-efficient computing platform for future medical imaging technology, which paves the road for low-power portable and point-of-care diagnostics. It also helps widely extend the application of memristor-based CIM beyond ANNs.x/HfO2/TiN. 30nm-thick TiN was sputtered as the top and bottom electrodes. 8nm-thick HfO2 was deposited by atomic layer deposition (ALD) as the resistive switching layer. 45 nm-thick TaOx was sputtered as the thermal enhanced layer to improve the analog switching characteristics45. As for the programming of 1T1R memristor array, we use the standard write-verify programming scheme45. Here, multiple voltage pulses are applied to the 1T1R memristor cell to increase (decrease) the conductance, until the conductance is larger  or equal to the target values. This process is repeated until the memristor conductance to programmed within the error margin of the target value.The memristor array was fabricated with a standard 0.13μm Si CMOS process. Each 1T1R cell consists of a memristor and a Si transistor. The memristor device has a material stack of TiN/TaOMathematically, DFT converts a complex Here, When mapping mentclass2pt{minimTo perform the computation of Eq.  on MIR, In this manner, 1D DFT is completed. The computing paradigm is shown in Fig. According to our experimental results shown in Supplementary Fig. To comprehensively evaluate the non-ideal device characteristics besides the mapping error, we run two simulations to analyze the impact of read noise and the stuck-at faults, respectively. As for the read noise, ten random scenarios with the read noise of 50-600 nA  are simulated. As shown in Supplementary Fig. 46, that the wire resistance does not have a serious impact on the computation accuracy for the 0.13μm technology node, and it starts to dominate the computation accuracy when the technology node is advanced to 14 nm or 28 nm. In such cases, techniques such as error balancing, bootstrapping and IR-drop adaptive compensation, can be used to mitigate the impact of wire interconnect resistance .The memristor array was fabricated with a standard 0.13μm Si CMOS process and the wire resistance between two adjacent memristors is estimated to be ~0.1Ω, based on the PDK information provided by the foundry. It is concluded in our previous workThe expression of 1D IDFT is given as:Separating the real and imaginary parts, Eq.  can be rWhen mapping on the memristor array, the computation can be performed as follows:Therefore, IDFT can then be implemented on MIR in a similar fashion as DFT, with only a minor difference on the sign of the imaginary part of DFT matrix.In addition, 2D DFT and 2D IDFT for an As shown in Supplementary Fig. The left atrium MRI dataset used in our work is one of the ten datasets in the Medical Segmentation Decathlon. The size of each MRI image is 320 × 320 and each dataset contains dozens of images. We randomly choose No.16 dataset to demonstrate the complete MRI reconstruction and segmentation. 19 other datasets  are used to further prove the consistent performance of MIR, as shown in Fig. 50 to convert CT images into simulated projections and choose the reconstruction algorithm based on Fourier central slice theorem51. Technically, we first divide every CT image into several patches of 36 × 36. Then we perform the Radon transform from 180 angles in MATLAB, and the length of each projection vector is 64. For the 64 × 180 projection matrix, 64-point DFT is implemented on MIR. The transformed output vector of each projection vector is 64-point long and is then filled into a 64 × 64 2D Fourier space. After the 64-point 2D IDFT implemented on MIR, the 64 × 64 2D Fourier space is converted into a 64 × 64 spatial-domain image. At last, the 64 × 64 image is cropped into a 36 × 36 patch as a part of the reconstructed CT image. In addition, an overlap strategy is also used in CT reconstruction to obtain the best reconstruction quality. Here, we use a famous spleen CT image dataset from Memorial Sloan Kettering Cancer Center. Each dataset contains about 100 slices and the size of each slice is 512 × 512. The pixel value represents the Hounsfield Unit (HU) value. More details of CT scan criteria can be found in ref. 52.To simulate the actual CT image reconstruction process, we perform ideal Radon transformhttps://github.com/MIC-DKFZ/nnUNet) runs on Python 3.8.8 with PyTorch framework 1.7.1. To perform left atrium and spleen segmentations, we take the pretrained models for 3D semantic image segmentation with nnU-Net (can be found on Zenodo https://zenodo.org/record/3734294).In this work, we implement nnU-Net on a GeForce GTX 1080 Ti GPU server with CUDA compilation tools 10.1. The nnU-Net’s source code 53 are:I represents the maximum pixel value of image I. The mathematical expression of SNR is:We use two widely accepted metrices, PSNR and SNR, to evaluate the fidelity of reconstructed images. The definition of PSNR is related to mean squared error (MSE). Given an ideal 54:X and Y represent two different datasets (segmented results).DICE score, ranging from 0 to 1, is widely used to measure whether two sets of data match well. Here, for the segmentation tasks, 1 corresponds to a perfect match between the segmented results and the ground truth, while 0 corresponds to no match. The expression of DICE score is given as follows55 using 65 nm technology). The transpose operation is reasonably neglected here because of its low computational cost. Thus, the overall number of required operations for one MRI image reconstruction is 640 × 320 × 320 × 2 × 4 = 524,288,000, and the energy efficiency of MIR in MRI reconstruction task is 10.9 TOPS · W−1 . In comparison, the energy efficiency of representative Intel 12th-Gen i9-12900 CPU and Nvidia Tesla V100 GPU is 9.5 GOPS · W−1  and 100 GOPS · W−1  and our MIR shows 1190In the MRI image reconstruction task, an MRI raw data matrix with a size of 320 × 320 is used as the standard input data for estimating the energy efficiency. In total, 640 320-point IDFTs are carried out on MIR, consuming 47.9 μJ , showing 1610 × and 153 × times advantages over Intel 12th-Gen i9-12900 CPU  and Nvidia Tesla V100 GPU , respectively.In the CT image reconstruction task, a 768 × 180 projection matrix is used as the standard input raw data. First, 180 768-point DFTs are carried out on MIR. This step consumes a total of 57.0 μJ . As for Nvidia Tesla V100 GPU, which could deliver a computing power of 31.4 TOPS, with a large die area of 815 mm2, the computational density is estimated as 39 GOPS · mm−2 . Therefore, in terms of the normalized image reconstruction speed, our MIR for MRI shows about 471Then we still choose Intel 12th-Gen i9-12900 CPU and Nvidia Tesla V100 GPU for comparison. As for Intel 12th-Gen i9-12900 CPU, it could provide a computing power of 0.6 TOPS and it takes an area of 200 mmSupplementary InformationPeer Review File


The benefits of access to blue spaces (exposure to aquatic environments) have been well reported. One common activity conducted in these spaces is recreational angling. Studies have shown that several correlates are associated with recreational angling, including a lower incidence of anxiety disorders compared to non-anglers. What is currently unknown is how measures of social support relate to feelings of loneliness in this population. The aim of this study, therefore, is to examine experiences of loneliness and social support in male UK anglers. In total, 1752 participants completed an online survey. The results of this study showed that the higher the number of friends or family that anglers hear from and feel close to, the less likely they are to report a lack of companionship, the less likely they are to report feeling left out, and the less likely they are to report feelings of isolation. Furthermore, more than half of the sample reported hardly ever or never having feelings of loneliness, suggesting that recreational angling does not affect feelings of loneliness. The benefits of access to blue spaces  are well reported. For example, one systematic review including 35 studies reported positive associations between exposure to blue spaces and mental health, overall wellbeing, and levels of physical activity . The rev−‘Considering the people you are related through birth, marriage, adoption and so on, how many relatives do you see or hear from at least once a month?’−‘Considering the people you are related through birth, marriage, adoption and so on, how many relatives do you feel at ease with that you can talk about private matters?’−‘Considering the people you are related through birth, marriage, adoption and so on, how many relatives do you feel close to such that you could call on them for help?’−‘How many of your friends do you see or hear from at least once a month?’−‘How many friends do you feel at ease with that you can talk about private matters?’−‘How many friends do you feel close to such that you could call on them for help?’The full methods of this study have been fully published elsewhere . In brie−‘How often do you feel that you lack companionship?’−‘How often do you feel left out?’−‘How often do you feel isolated from others?’Questions about loneliness included questions regarding perceived companionship, feeling left out, and feeling isolated. Specifically, these three nominal questions were asked: Participants read a participant information sheet and subsequently provided informed consent prior to the completion of the survey. Ethical approval for the present study was granted by the Anglia Ruskin University Sport and Exercise Science Ethics Panel. All statistical analyses were conducted using SPSS Version 28. 0.0.0 (190) . Data from the social correlate variables were analyzed using a descriptive analysis. To determine the associations between social correlates , a Mantel–Haenszel test of trend was performed. n = 40). It was therefore deemed appropriate to only include the male sample. In total, 1752 male participants completed the online questionnaire. The majority of participants were either aged between 55 and 64 (25.8%), 45 and 54 (20.8%), or 65 and 74 (20.4%). Originally, a total of 1792 anglers completed the online survey; however, of this sample there was a very small number of female respondents , or 5 to 8 (27.8%) relatives. When the same question was asked about how many friends they heard from at least once a month, the majority of participants had also heard from 3 to 4 (29.6%), or 5 to 8 (18.9%) friends. Regarding how many relatives participants felt at ease with that they could talk about private matters, the majority of participants stated 3–4 (27.5%), followed by 1 (22.1%). When the same question was asked regarding friends, the results were similar, with 27.6% of participants saying that they had 3–4 friends they could talk to about private matters. Regarding how many relatives they felt close to such that they could call on them for help, the majority of participants indicated 3–4 (31.9%), followed by 2 (20.7%). Results were similar when asked the same question about friends that they could call on for help. See Regarding companionship, most participants (57.5%) stated that they hardly ever or never felt a lack of companionship. Regarding feeling left out, 55.2% stated that they hardly ever or never felt left out. Lastly, regarding isolation, the majority of participants (57.2%) stated that they hardly ever or never felt isolated from others. 2(1) = 85.80, p < 0.001, and r = −0.22). A similar trend was also found regarding the association between how many friends recreational anglers saw or heard from once a month and how often they felt a lack of companionship (χ2(1) = 92.96, p < 0.001, and r = −0.23). A significant trend was also found between how many relatives recreational anglers felt they could talk about private matters with and how often they felt a lack of companionship (χ2(1) = 117.40, p < 0.001, and r = −0.32), with a similar trend found for friends (χ2(1) = 117.40, p < 0.001, and r = −0.29). Lastly, a significant trend was also found between how many relatives recreational anglers felt that they could call for help and how often they felt a lack of companionship (χ2(1) = 207.52, p < 0.001, and r = −0.35), with a similar trend being found for friends (χ2(1) = 208.91, p < 0.001, and r = −0.35). The Mantel–Haenszel test of trend between how many relatives recreational anglers saw or heard from once a month and how often they felt a lack of companionship showed a statistically significant linear association (χ2(1) = 94.79, p < 0.001, and r = −0.23). A similar trend was also found regarding the association between how many friends recreational anglers saw or heard from once a month, and how often they felt left out (χ2(1) = 122.73, p < 0.001, and r = −0.27). A significant trend was also found between how many relatives recreational anglers felt they could talk about private matters with and how often they felt left out (χ2(1) = 197.71, p < 0.001, and r = −0.34), with a similar trend found for friends (χ2(1) = 184.42, p < 0.001, and r = −0.33). Lastly, a significant trend was also found between how many relatives recreational anglers felt that they could call for help and how often they felt left out (χ2(1) = 203.51, p < 0.001, and r = −0.34), with a similar trend being found for friends (χ2(1) = 212.61, p < 0.001, and r = −0.35). A Mantel–Haenszel test of trend between how many relatives recreational anglers saw or heard from once a month, and how often they felt left out showed a statistically significant linear association (χ2(1) = 129.48, p < 0.001, and r = −0.27). A similar trend was also found regarding the association between how many friends recreational anglers saw or heard from once a month, and how often they felt isolated (χ2(1) = 156.73, p < 0.001, and r = −0.30). A significant trend was also found between how many relatives recreational anglers felt they could talk about private matters with and how often they felt isolated (χ2(1) = 245.46, p < 0.001, and r = −0.35), with a similar trend found for friends (χ2(1) = 191.83, p < 0.001, and r = −0.33). Lastly, a significant trend was also found between how many relatives recreational anglers felt that they could call for help and how often they felt isolated (χ2(1) = 241.42, p < 0.001, and r = −0.37), with a similar trend being found for friends (χ2(1) = 230.39, p < 0.001, and r = −0.36).A Mantel–Haenszel test of trend between how many relatives recreational anglers saw or heard from once a month, and how often they felt isolated showed a statistically significant linear association (χThis study examined trends between social support correlates and feelings of isolation in a cohort of male recreational anglers. The results of this study showed that the higher the number of friends or family that anglers hear from and feel close to, the less likely they are to report a lack of companionship, the less likely they are to report feeling left out, and the less likely they are to report feelings of isolation. These findings are in agreement with previous studies that have found similar associations between social support and loneliness in general populations ; howeverFindings from this study should be interpreted in light of its limitations. Firstly, because of the cross-sectional nature of the study, and the impossibility of determining dependent and independent variables in the Mantel–Haenszel test, causal inferences could not be made. Future longitudinal cohort studies are now required. Secondly, the methods of recruiting the sample of recreational anglers could be open to potential selection bias. For example, the use of an online survey could exclude groups who are less likely to have internet access or access to social media. Therefore, populations with lower incomes, with lower levels of computer literacy, and older adults may be under-represented in this study. Thirdly, the study variables were self-reported and thus potentially introduced social desirability and recall bias into the study findings. Finally, very few females responded to the online survey and thus were removed from the present analysis. Future research in the female angling population is now required. In conclusion, in the present study including a sample of 1752 male recreational anglers from the UK, it was found that those who reported having more friends and family that they hear from, can call upon for support, and trust enough to talk to about private matters are less likely to report feelings of a lack of companionship, feelings of being left out, and feelings of isolation. Furthermore, a significant proportion of recreational anglers reported having very few feelings of lack of companionship, feelings of being left out, and feelings of isolation, despite having a sometimes-solitary hobby. Future longitudinal cohort studies are now required before any concrete recommendations can be provided. Moreover, it would be prudent to repeat the present study in a representative sample of female anglers to determine whether the findings also apply to females.


Aim: To systematically assess and rank the efficacy of opioid medications for traumatic pain in the emergency department in terms of pain relief, adverse events and rescue analgesia.Methods: Four databases were systematically searched until 26 September 2022: PubMed, Embase, Cochrane Library, and Web of Science. Outcomes were pain relief, adverse events , and rescue analgesia. For each outcome, network plots were drawn to exhibit direct and indirect comparisons, and rank probabilities were utilized to rank the efficacy of different opioids.Results: Twenty studies of 3,040 patients were eligible for this network meta-analysis. According to the rank probabilities, the top three analgesic medications for pain relief may be sufentanil (78.29% probability of ranking first), buprenorphine (48.54% probability of ranking second) and fentanyl (53.25% probability of ranking third); buprenorphine (31.20%), fentanyl (20.14%) and sufentanil (21.55%) were least likely to cause dizziness; the top three analgesic medications which were least likely to cause hypotension were buprenorphine (81.64%), morphine (45.02%) and sufentanil (17.27%); butorphanol (40.56%), morphine (41.11%) and fentanyl (14.63%) were least likely to cause pruritus; the top three medications which were least likely to cause sedation were hydrocodone + acetaminophen (97.92%), morphine (61.85%) and butorphanol (55.24%); patients who received oxycodone (83.64%), butorphanol (38.31%) and fentanyl (25.91%) were least likely to need rescue analgesia in sequence.Conclusion: Sufentanil, buprenorphine and fentanyl may be superior to other opioid medications in terms of pain relief and the incidence of dizziness, hypotension and pruritus, which might be selected as opioid analgesics for traumatic pain in the emergency setting. Pain is a common complaint among trauma patients in the emergency department . Up to 7versus those using tramadol only  independently: PubMed, Embase, Cochrane Library, and Web of Science. The last search was conducted on 26 September 2022. English search terms were as follows: “Opioid” OR “Opioids” OR “Opiate” OR “Opiates” OR “Opium” OR “Papaveretum” OR “Omnopon” OR “Pantopon” OR “Morphine” OR “Morphia” OR “MS Contin” OR “Oramorph SR” OR “Duramorph” OR “Hydromorphone” OR “Dihydromorphinone” OR “Hydromorphon” OR “Dilaudid” OR “Hydrocodone “OR “Hydrocodon” OR “Dihydrocodeinone” OR “Dicodid” OR “Hycodan” OR “Oxymorphone” OR “Numorphan” OR “Oxycodone” OR “Dihydrone” OR “Eucodal” OR “Oxycontin” OR “Fentanyl” OR “Fentanil” OR “Phentanyl” OR “Fentanest” OR “Sublimaze” OR “Fentora” OR “Sufenta” OR “Heroin” OR “Meperidine” OR “Pethidine” OR “Dolosal” OR “Dolcontral” OR “Dolantin” OR “Demerol” OR “Lidol” OR “Lydol” OR “Operidine EPJ I” OR “Levorphanol” OR “Levorphan” OR “Codeine” OR “Tramadol” OR “Tramundin” OR “Zydol” OR “Adolonta” OR “Contramal” OR “Methadone” OR “Dolophine” OR “Metadol” OR “Symoron” OR “Methadose” OR “Phenadone” OR “Physeptone” OR “Amidone” OR “Methaddict” OR “Buprenorphine” OR “Buprenex” OR “Subutex” OR “Pentazocine” OR “Talwin” OR “Fortral” OR “Butorphanol” OR “Moradol” OR “Stadol” OR “Torbugesic” OR “Nalbuphine” OR “Nubain” OR “Dezocine” AND “acute trauma*” OR “trauma pain*” OR “traumatic pain*” OR “traumatic injur*” OR “acute pain*” OR “acute injur*” OR “acute wound*“. The references of relevant meta-analyses were manually retrieved to avoid missing qualified studies. Titles and abstracts of the retrieved studies were screened primarily, and full texts were used for subsequent screening. This network meta-analysis was performed following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) extension statement for network meta-analyses .Inclusion criteria included: 1) studies on patients with traumatic pain in the emergency department (population); 2) studies on opioids such as morphine, fentanyl, tramadol, oxycodone, sufentanil, and codeine (intervention and comparator); 3) studies on the following outcomes: pain relief, adverse events , and rescue analgesia (outcome); 4) RCTs, cohort studies, and non-randomized controlled trials (study design). Pain relief was defined as the difference in pain scores between before treatment and after treatment, indicating the degrees of pain reduction.Exclusion criteria included: 1) studies on a mixed population, such as a population of trauma patients and patients with other acute pain; 2) studies with a control group using non-opioid drugs alone, such as acetaminophen, ketamine, lidocaine, and ibuprofen; 3) studies on animal experiments; 4) studies with incomplete or un-extractable data; 5) case reports, meeting abstracts, letters, reviews, meta-analyses; 6) non-English publications.Two independent authors (YW Fu and Q Liu) extracted the following data from the included studies: first author, year of publication, country, study design, study population (indication), drug administration and type, dose, number of participants, sex , age (years), pain score, trauma type, follow-up time (FU), and outcome. Disagreements were resolved through discussion with another author (H Nie).For the quality evaluation of RCTs, the Cochrane Collaboration’s tool for risk of bias assessment was used . Six dom2 statistic was the main indicator of statistical heterogeneity, with values < 25%, 25%–50% and >50% signifying low, moderate and high heterogeneity, respectively. Consistency referred to the statistical consistency between direct and indirect effect sizes for the same comparison. The deviation information criterions (DICs) of the consistency model and the non-consistency model were compared, and a small value suggested a better fit. The absolute value of the difference in the DICs within 5 denoted consistency between indirect and direct evidence.The network meta-analysis allows assessment of multiple interventions in a single analysis via combining direct and indirect evidence. Direct evidence means evidence from studies directly comparing interventions A and B (head-to-head comparison) and is an estimate of relative effects between A and B. Indirect evidence refers to the evidence obtained from paths with one or more common comparators. For instance, when there are no studies that directly compare A and B (direct evidence), an indirect comparison between A and B can be made if both are compared with C in studies (indirect evidence) . This neNetwork plots were drawn to exhibit direct and indirect comparisons for each outcome. A larger node indicated a larger sample size for the medication represented by the node, while a thicker line indicates a larger number of studies for comparison of the medications at both ends of the line. Forest plots and league tables were depicted to illustrate the effects of opioids on the outcomes. For pain relief, weighted mean differences (WMDs) and 95% credibility intervals (CrIs) were reported; for adverse events and rescue analgesia, relative risks (RRs) and 95%CrIs were displayed. Rank probabilities were utilized to rank the efficacy of different opioids for each outcome. STATA 15.1 , RevMan 5.4  and R 4.1.3  software was adopted for statistical analysis.A total of 19,720 were retrieved from PubMed , Embase , Cochrane Library , and Web of Science . After deleting duplicates , and excluding ineligible studies based on titles and abstracts  and on full texts (n = 111), 20 studies  of 3,040For 17 RCTs, the overall risk of bias was low. The effect of opioid medications on pain relief was investigated in 13 studies of 2,225 patients, involving 11 medications: buprenorphine, odeine + acetaminophen, fentanyl, fentanyl + tramadol, hydrocodone + acetaminophen, morphine, oxycodone, oxycodone + acetaminophen, sufentanil, tramadol, and tramadol + acetaminophen. Morphine and hydrocodone + acetaminophen were used in more patients than other medications. There were more studies for direct comparisons between morphine and fentanyl, and between oxycodone + acetaminophen and hydrocodone + acetaminophen .According to the forest plot, sufentanil relieved significantly more pain than morphine  . The leaThe incidence of dizziness after the following 10 opioid medications was explored by 11 studies with 1,265 patients: buprenorphine, codeine + acetaminophen, fentanyl, fentanyl + tramadol, hydrocodone + acetaminophen, morphine, oxycodone, oxycodone + acetaminophen, sufentanil, and tramadol. The direct comparison of morphine and fentanyl was conducted in more studies. Morphine and hydrocodone + acetaminophen were used in more patients .No significant difference was found between the investigated opioid medications, according to the forest plot  and leagThree studies with 293 patients provided data on the influence of four analgesics on hypotension: morphine, sufentanil, oxycodone, and buprenorphine. Morphine was the most commonly studied analgesics. Morphine was directly compared with other three drugs .versus buprenorphine administration . Oxycodone was associated with a significantly increased incidence of hypotension compared with buprenorphine  . As shown in the league table, the incidence of hypotension was significantly higher after morphine administration , 99.93) . The ran, 99.93) .The impact of opioid medications on pruritus was evaluated in 6 studies of 1,048 patients, and 6 medications were involved: buprenorphine, codeine + acetaminophen, fentanyl, hydrocodone + acetaminophen, morphine, and oxycodone + acetaminophen. Morphine was directly compared with fentanyl in more studies, and more patients were administrated morphine and oxycodone + acetaminophen .The forest plot exhibited that patients using oxycodone + acetaminophen had a significantly greater incidence of pruritus than those using morphine  . In the Four studies of 368 patients assessed the effect of butorphanol, fentanyl, hydrocodone + acetaminophen, and morphine on sedation. Morphine was the most commonly studied opioid, and it was directly compared with the other three analgesics .Patients using hydrocodone + acetaminophen had a significantly lower incidence of sedation compared with those using morphine, as displayed by the direct comparison result of the forest plot . The league table found that morphine was associated with a significantly elevated incidence of sedation than hydrocodone + acetaminophen . AccordiData on rescue analgesia were provided by 11 studies with 1,755 patients, involving 8 opioid medications: butorphanol, codeine + acetaminophen, fentanyl, hydrocodone + acetaminophen, morphine, oxycodone, oxycodone + acetaminophen, and sufentanil. More studies investigated the direct comparison between morphine and fentanyl, and more patients used these two opioids .The forest plot demonstrated no significant difference between the presented medications . AccordiIn the current network meta-analysis, the efficacy of opioid medications for traumatic pain in the emergency department was first systematically evaluated as regards pain relief, adverse events and rescue analgesia. We found that sufentanil, buprenorphine and fentanyl may be more effective in relieving pain, and may relate to a lower incidence of dizziness, hypotension and pruritus, and need for rescue analgesia among opioid medications for trauma patients, suggesting that sufentanil, buprenorphine and fentanyl may have advantages over other opioid medications in treating traumatic pain, which might be chosen as opioid analgesics in the emergency department.versus fentanyl and morphine may lead to accidental overdose of sufentanil (A network meta-analysis was recently performed by Yin et al.  to assesfentanil . Sufentafentanil . Murray fentanil  showed tfentanil . Recentlfentanil . Fentanyfentanil . It has fentanil . This neConcerning adverse events, the incidence of dizziness after sufentanil, buprenorphine and fentanyl may be lower than that after the other opioid medications based on the result of the rank probability, although no significant difference were shown. For hypotension, only four opioids were evaluated, and buprenorphine may be associated with the lowest incidence of hypotension, while fentanyl was not investigated in the qualified studies. For pruritus and sedation, sufentanil and buprenorphine were not assessed by the included studies, and patients using fentanyl may have the third lowest incidence of pruritus but an unfavorable incidence of sedation. More studies are needed in the future to verify the role of different opioids in consideration of adverse events. Of note, the proportion of patients who required rescue analgesia may be relatively low after receiving fentanyl, and relatively more patients may require rescue analgesia after sufentanil treatment, which necessitated large-scale studies for corroboration.Based on our findings from this comprehensive assessment with 20 studies of 3,040 patients, sufentanil, buprenorphine and fentanyl may provide favorable effects on traumatic pain relief, and may be associated with a lower incidence of dizziness, hypotension and pruritus, and need for rescue analgesia, which might serve as a reference in choosing opioid medications for traumatic pain management in the emergency department. Some limitations should be noted in interpreting the findings. Firstly, the dosage of same medications varied across different studies, which precipitated increased study heterogeneity. Secondly, equipotent doses were not studied given the clinical limitations of care of the trauma research subjects enrolled in many of the studies, therefore higher and lower doses were titrated to clinical effect. If the doses compared were not equipotent, this may affect safety and efficacy in traumatic pain. For example, higher doses might provide more pain relief. Besides, the small sample size of the included studies may affect the stability of our results. Thirdly, different administration routes of the same medication may affect the analgesic effect, and subgroup analysis based on administration routes could not be achieved due to limited eligible studies. As shown in For trauma patients with musculoskeletal or trauma-related pain in the emergency department, sufentanil, buprenorphine and fentanyl may provide favorable effects on pain relief, and may be associated with a lower incidence of dizziness, hypotension and pruritus, and need for rescue analgesia among opioid medications. Confirmation of these finding requires further studies.


Previous in vitro characterisation of the complex has led to the hypothesis of a complex involved in iron metabolism and control of c-di-GMP levels. We demonstrate that the stressosome is probably involved in reshaping the glucose metabolism in Fe- and nutrient-limited conditions and mutations of the locus affect the activation of the glyoxylate shunt. Moreover, we show that the stressosome is needed for the transcription of fleQ and to promote motility, consistent with the hypothesis that the stressosome is involved in regulating c-di-GMP. This report highlights the potential role of the stressosome in a Gram-negative bacterium, with implications for the metabolism and motility of this pathogen.Stressosomes are signal-sensing and integration hubs identified in many bacteria. At present, the role of the stressosome has only been investigated in Gram-positive bacteria. This work represents the first   Transcrer (ASW) . More rebacteria . Despite sensing .V. vulnificus. The two mutants produce similar phenotypes indicating that there is a functional link between the two modules. We demonstrate that the stressosome is expressed and is active in a nutrient-limited environment. We also show that the stressosome is most likely involved in remodelling metabolism in V. vulnificus in this environment, through activation of the glyoxylate shunt, an alternative to the TCA cycle that allows acclimation to low iron/nutrient conditions in several microorganisms  medium with additional 0.4 M NaCl (LBN) or Chemically Defined Medium (CDM) -Glucose, 0.75 μM FeCl3) at 37 °C. The FeCl3 stock was prepared in 1 M HCl. E. coli β2163 was grown in LB medium supplemented with 0.3 mM 2,6-Diaminopimelic acid (DAP) at 37 °C. Overnight broth cultures were grown in 2 ml or 20 mL medium, in 15 mL bacterial culture tubes and 250 mL flasks, respectively, at 37 °C with agitation at 150 rpm. Specific growth conditions, different from the one indicated above, are described in the appropriate sections. All chemicals and reagents were supplied by Sigma-Aldrich, unless indicated otherwise.The clinical isolate us CMCP6  was usedjugation . Vibrio 2.2VvrsbR, VvrsbS, VvrsbT and VvrsbX genes (V. vulnificus ΔRSTX) and one lacking the first gene of the downstream module VvD1 (V. vulnificus ΔD1), encoding for the sensor kinase of a two-component regulatory system, predicted to be involved in cyclic-di-GMP regulation. Construction of the upstream ΔRSTX mutant has been previously described  were grown overnight at 37 °C in LBN and LB + 25 μg/mL Chloramphenicol + 0.3 mM DAP, respectively. Cultures were washed by centrifugation in LB broth, the two strains were mixed at a 1:1 ratio (v/v), spotted onto LB + 0.3 mM DAP agar plates and incubated for 7 h at 37 °C. V. vulnificus transconjugants carrying the knock-out plasmid integrated into the chromosome by homologous recombination were selected on LBN + 5 μg/mL Chloramphenicol. In order to promote the excision of the plasmid from the chromosome of V. vulnificus, the second event of homologous recombination was favoured by culturing the first recombinant cells in LBN broth without selective antibiotic. Second recombinant cells were selected on LBN + 5% sucrose (w/v) because the sacB gene present in the suicide vector pDS132 does not allow growth in the presence of sucrose and growth on this media is a sign of a successful excision of the plasmid from the bacterial genome. This event can equally generate wild-type cells or deletion mutants. For this reason, mutants were selected by PCR screening of colonies grown on LBN + 5% sucrose using Taq polymerase (Bioline) and the primers D1_For (5′-GATGGGGCTGTCATTCACGA-3′) and D1_Rev (5′-TCATGCAAAGGGGCAGACTT-3′). PCR products of different sizes for the mutant and the wild-type were visualised through agarose gel electrophoresis. One mutant was selected and Whole Genome Sequencing (WGS) analysis was performed. Genomic DNA was extracted using the Wizard Genomic DNA Purification Kit (Promega), according to the manufacturer's protocol. Genomic DNA was sequenced by MicrobesNG  using Illumina technology. The average read length was 190 nucleotides for each sample and average fold coverage was 100. BreSeq version 0.32.0 was used to call base substitution mutations Read Alignment evidence using Consensus mode, with a mutation E-value cut-off of 10 and frequency cut-off of 0.8 (80%).Two knock-out mutants were used in this work, one lacking the upstream module of the stressosome locus consisting of the escribed . Construe vector , betweenli β2163  allows c species . V. vuln2.3600 of 0.05 in 250 mL flasks. The flasks were incubated at 37⁰C with 150 rpm agitation. OD600 was measured every 60 min for the first 12 h and at different time points  between 24 and 72 h.Growth was assessed for all the strains at 37 °C in CDM and LBN. The strains were grown overnight in 20 mL LBN at 37 °C with at least three biological replicates. Bacterial cultures were then washed and diluted in 30 mL of appropriate media to a final OD2.4V. vulnificus WT and stressosome mutants in CDM at 37 °C, 1 mL culture was taken from the culture flask at different time points and filtered with a 0.22 µm filter for downstream HPLC analysis. Samples were stored in a tightly closed tube at −20 °C. The HPLC analysis was performed in a liquid chromatograph (LC)  with a refractive index detector (RID) and a Hi-Plex H column (300 × 7.7 mm) at 60 °C. 5 mM H2SO4 was used as mobile phase and the flow rate was set at 0.4 mL/min. For each time point three replicates were analysed.In order to study the glucose depletion and organic acid accumulation in the media during growth of 2.5600 of cells, was mixed with 2 vol of RNAprotect (Qiagen) and processed according to the manufacturer's instructions. Treated pellets were stored for a maximum of 24 h at −20 °C. Before RNA extraction, each pellet was resuspended in 200 µL TE buffer  + 15 mg/mL lysozyme + 10 µL ready-to-use QIAGEN Proteinase K (20 mg/mL) and incubated 10 min at room temperature to lyse the bacterial cells. Lysates were used for RNA extraction with RNeasy Mini Kit (Qiagen) following the manufacturer's protocol. RNA was eluted in 50 µL RNase-free water and a rigorous DNase treatment was performed using the TURBO DNA-free kit (Life Technologies), according to the manufacturers’ instruction, to remove traces of DNA. RNA concentration and quality were assessed with a NanoDrop spectrophotometer and only samples with a 260/280 ratio ≥ 1.7 were used for the qRT-PCR protocol, and RNA integrity was verified on a 1.5% (w/v) agarose gel. cDNA was prepared from 1 µg total RNA using the Transcriptor First Strand cDNA Synthesis Kit (Roche) and Random Hexamer Primer contained in the kit. The RT reaction was carried out using a 10 min incubation at 25 °C followed by 60 min at 50 °C. The primers for the qPCR reactions (tuf gene (elongation factor Tu) as endogenous control, similarly to previously-published transcriptional studies in V. vulnificus  and used to determine the number of CFU/mL, through ten-fold serial dilutions and colony counting on LBN agar. Three biological replicates were tested for each strain, each one plated in duplicate.To test survival ability in CDM at pH 5, the three strains were grown in 2 mL LBN broth for a minimum of 17 h. Each culture was then washed and diluted to OD2.7In order to determine swimming on CDM, the three strains were grown overnight in CDM broth. Plate counting was performed on overnight cultures as previously described to verify that all the strains were viable after overnight growth in CDM. A sterilized metal wire was dipped in the overnight culture and used to pierce the centre of the CDM motility plates (agar concentration 3.35 g/L). Plates were incubated at the appropriate temperature and the motility zone was measured after 16 h. Three biological replicates were tested for each strain.2.8V. vulnificus CMCP6 and the two stressosome mutants, strains were grown in biological triplicates overnight in CDM broth. Plate counting was performed on overnight cultures to verify that all the strains were viable after growth in CDM. The overnight culture was then diluted in fresh CDM broth at a final OD600 of 0.1 and 200 µL cell suspension, assessed in three technical replicates, added to wells of a microtiter plate. The plates were incubated statically at 30 °C or 37°⁰C for 24 h and biofilm quantified through crystal violet staining . Bacterial cultures were first diluted to an OD600 of 0.01 in PBS and then added to the HeLa cells to achieve an MOI of 10. The plate was incubated at 37 °C and HeLa cells lysis was determined at different time points, through Lactate Dehydrogenase (LDH) quantification, using CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega), according to the manufacturer's protocol. HeLa cells treated with 1X Lysis buffer provided by the manufacturer were used as the positive control (100% lysis). HeLa cells in the absence of bacteria were used as the negative control (0% lysis). For each strain at least two biological replicates, each one assessed with technical triplicates, were analysed in at least two independent experiments.To study cytotoxicity of olyticus . HeLa ce2.10To test the production of protease, all strains were grown overnight in LBN broth and inoculated onto LBN plates containing 1% (w/v) skim milk clearing zones were measured after 24 h incubation at 37 °C.33.1V. vulnificus has so far not been characterised in vivo. To determine the conditions in which the stressosome may be physiologically active, transcriptional analysis of the upstream  and downstream  modules of the stressosome locus , which represents periods of stress. Effects on V. vulnificus wild-type growth of different iron concentrations were investigated and the concentration of 0.75 µM was identified as growth-limiting and used in this work . For the transcriptional analysis, bacteria were cultured for 16 h in LBN and, following the shift from nutrient-rich media (time point 0 h) to CDM, transcription was quantified at 6, 12 and 24 h  as reference sequences and by comparing the mutants with the wild-type. The WGS confirmed the knockout mutations and excluded the presence of secondary mutations shared between the two mutants or other mutations that could explain the phenotypes presented in this work (data available upon request). Growth characterisation of both mutants in LBN showed no differences in growth rate and overall biomass accumulation between these and the wild-type strain . Moreover, an extensive phenotypical characterisation of the ΔRSTX mutant in LBN showed no differences caused by the mutation. The mutant was tested for growth, survival and tolerance to several stresses, cross-protection and motility in rich media and showed physiological traits identical to the wild-type strain .As shown above, the stressosome is upregulated during growth in CDM, when glucose is depleted and acetic acid produced. For this reason, phenotypic characterisation of the ΔRSTX and ΔD1 strains was performed in CDM in this study. The effects of each mutation on the transcription of the genes in the other module were tested through qRT-PCR.Deletion of the upstream module seemed to reduce transcription of the downstream V. vulnificus in CDM. The growth measurements revealed a partial growth advantage of ΔD1 in the late-log and early-stationary phase  and accumulates acetate, to a pathway, generally characterised by a lower growth rate, that facilitates the utilisation of that acetate that was previously accumulated. The acetate switch has been widely studied and can be considered as a survival pathway that occurs when the principal carbon source has been depleted , whether metabolic pathway alternatives to the glycolysis/TCA cycle were differentially regulated in the mutants when compared to the wild-type during growth in CDM. Several intermediates of the glucose central metabolic pathway can be utilised for alternative metabolic pathways, such as acetate metabolism, glyoxylate shunt and gluconeogenesis, based on various environmental factors including oxygen levels, iron availability and nature and quantity of the carbon source . As prevdepleted . In thisV. vulnificus is that it functions as an iron-sensing complex (2 in the TCA cycle), through the production of oxaloacetate and subsequent gluconeogenesis (aceA (encoding isocitrate lyase) and aceB , was tested together with the glcB gene encoding a malate synthase-related protein (glcB gene) following the shift from LBN to CDM, similar to the one observed in the ΔD1 strain. Each of the analysed genes was upregulated at subsequent time points although with lower transcription levels than the wild-type. Thus during the stationary growth phase, gene transcription in the ΔRSTX strain followed a pattern similar to that of the wild-type.One of the previously hypothesised roles of the stressosome in  complex . Moreove complex . This meogenesis . The tra protein B and 2C.3.4V. vulnificus accumulates acetic acid and lactic acid during growth in CDM in the presence of glucose. The accumulation of organic acids causes a significant pH drop in our experimental conditions and this has been previously reported to cause loss of viability in several bacterial species through the so-called “suicide phenomenon” , so the extended growth profile was not due to alterations in the bacterial response to stress. However, after 72 h growth, the viability of the wild-type strain decreased more than the mutants  state. The results indicated that they were indeed dead as indicated by loss of membrane integrity in more than 95% of CMCP6 cells after 72 h growth, as determined by cellular influx of membrane-impermeable fluorescent dye. .Although a degree of variability was observed, the viability of the wild-type and mutants was similar after 48 h growth  was first identified in Pseudomonas aeruginosa as the master gene at the top of the hierarchy , and part of the downstream module formed by two genes (VvD1 and VvD2) encoding a two-component regulatory system most likely involved in the degradation of c-di-GMP and hypothesised to be the regulatory output of the stressosome in V. vulnificus  show a partial growth advantage in the late exponential phase and early stationary phase, at the time in which the transcription of the stressosome genes reaches its peak in the wild-type strain. Despite the higher growth, no differences in the rate of consumption of glucose or accumulation of organic acids were observed. To investigate the possible causes of this growth difference, the transcription of the genes involved in metabolic pathways alternative to the TCA cycle was tested. In particular, we targeted the acetate metabolism pathway  was down-regulated in ΔRSTX compared to the wildtype. Both approaches also demonstrated that acs (VV1_1237), pta (VV1_2220) and rpoS (VV1_1588) were expressed at similar levels in WT and ΔRSTX. While toxR (VV1_0190) and fleQ (VV1_1931) transcription was down-regulated in ΔRSTX, there was no change in protein levels compared to the WT, possibly due to continued protein stability despite reduced de novo synthesis. qRT-PCR studies demonstrated an intermediate effect of deleting RSTX on aceA (VV1_0449), aceB (VV1_0450) and glcB (VV2_1647) transcription compared to ∆D1 and WT. Likewise there was minimal influence of RSTX on AceA or AceB protein levels, though GlcB levels did decrease.Global expression analysis comparing proteomes of ublished . We compin vivo characterisation of the stressosome in V. vulnificus and in a Gram-negative bacterium. This work provides the first insight into the effects that this locus can have on the physiology of this human pathogen. In line with the observation of an iron-dependant stressosome previously published , under grant number GOIPG/2020/1420. Additional support was provided by the Irish HEA (Higher Education Authority) as part of a COVID-19 costed extension programme.This research was jointly supported by a Marie Skłodowska-Curie ITN Fellowship  within the seventh European Community Framework Program and by the Laura Cutugno: Conceptualization, Methodology, Investigation, Data curation, Validation, Formal analysis, Visualization, Writing – original draft, Funding acquisition. Borja Khatabi Soliman Tamayo: Methodology, Investigation, Formal analysis, Writing – review & editing. Piet N.L. Lens: Resources, Validation, Supervision, Writing – review & editing. Conor O'Byrne: Conceptualization, Methodology, Validation, Writing – review & editing, Funding acquisition. Jan Pané-Farré: Conceptualization, Methodology, Validation, Writing – review & editing, Supervision, Funding acquisition. Aoife Boyd: Conceptualization, Methodology, Validation, Resources, Data curation, Writing – review & editing, Supervision, Project administration, Funding acquisition.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


This study provides scientific insights for the application of canopy remote sensing diagnostics of strawberry photosynthetic physiological chilling injury in practical agricultural production.Chilling injury can adversely affect strawberry bud differentiation, pollen vitality, fruit yield, and quality. Photosynthesis is a fundamental process that sustains plant life. However, different strawberry varieties exhibit varying levels of cold adaptability. Quantitatively evaluating the physiological activity of the photosynthetic system under low-temperature chilling injury remains a challenge. In this study, we investigated the effects of different levels of chilling stress on twenty photosynthetic fluorescence parameters in strawberry plants, using short-day strawberry variety “Toyonoka” and day-neutral variety “Selva” as representatives. Three dynamic chilling treatment levels  and three durations  were applied to each variety. WUE, LCP, Y(II), qN,  Fragaria × ananassa Duch.) is known for its exquisite and delectable fruits, rich in beneficial substances such as vitamins, anthocyanins, flavonoids, minerals, and phenolic compounds, making it highly popular among consumers worldwide. It is considered one of the most important horticultural crops . Th. Th49]. Between 9:00 and 12:00, we evaluated strawberry leaf photosynthetic parameters using a portable photosynthesis system . Our focus was directed towards the central leaflet of the second or third robust functional leaf, commencing from the uppermost point of the strawberry plants . The instion (1) . A lightcy (AQE) ,53. The Using a portable modulated chlorophyll fluorometer , we conducted measurements of chlorophyll fluorescence induction kinetics on strawberry leaves from 9:00 to 11:00 . Three hIn the clear open-air conditions, we utilized a fiber optic hyperspectral spectrometer  to collect leaf reflectance radiance spectra and obtained the “whiteboard” reflectance radiance as the incident solar irradiance spectrum. The observation was conducted from 11:00 to 12:00. The instrument’s operating spectral range was 645–800 nm, with a spectral resolution of 1.4 nm. During the observations, the leaves and the “whiteboard” were placed horizontally parallel to the ground, and a fixed setup maintained the leaves perpendicular to the fiber optic probe. Each measurement was repeated twice. In this study, the solar-induced chlorophyll fluorescence (SIF) was extracted using the radiance-based 3FLD algorithm . This meIn the equation, We employed a portable hyperspectral imaging system  to capture the reflectance spectra of strawberry leaves. The observations were conducted indoors from 12:00 to 14:00, using two 75 W tungsten halogen lamps  as artificial light sources. The instrument’s spectral range was 375.88–1039.19 nm, and we selected the 256-band mode with a spectral resolution of 2.6 nm for this study. The instrument was positioned vertically over the observation target, and after performing reference panel reflectance calibration on the acquired data, 2 or 3 regions of interest (ROI) were selected within the observation area of each target leaf. The average spectra of ROIs were used as the reflectance spectra of the target leaves.Due to the narrow band range of hyperspectral data, the signal-to-noise ratio is lower and data stability is weaker compared to spectral data with broader band ranges. This results in apparent sawtooth patterns and severe random fluctuations near the starting and maximum wavelengths on the spectral curve. To address these challenges, we selected a relatively stable range of 410.49–990.07 nm from the original spectral data for our study. Furthermore, a non-linear mean filter was applied to reduce noise interference . After cp = 0.05) and principal component analysis (PCA), were conducted. All observed parameters were reported as “Mean ± SD” (mean ± standard deviation).For statistical analyses in this investigation, we employed IBM SPSS Statistics version 24 . One-way analysis of variance (ANOVA), accompanied by Duncan’s multiple comparison test  ; th indicator of Ak; k; k; th indicator ; Ak; Ak; In the equation, 2), root mean square error  and (13) [Wavelet transformation is an important signal processing method used to analyze digital data . It decoand (13) .(12)ψa,In the equation, We established hyperspectral inversion models for the comprehensive score of the strawberry photosynthetic system under chilling stress using four different machine learning algorithms: Backpropagation Neural Network (BPNN) , Random In experiments conducted at three dynamic chilling conditions of 20/10 °C, 15/5 °C, and 10/0 °C, with stress durations of 3, 6, and 9 days, the cold tolerance of short-day strawberry cultivar “Toyonoka” surpassed that of the long-day representative “Selva”. Under continuous treatment at 20/10 °C, after 6 days, different photoperiod strawberry types exhibited initial signs of cold-induced physiological stress and escalating stress levels evident by the 9th day, with “Selva” experiencing aggravated stress while “Toyonoka” rebounded to an unstressed state. At 15/5 °C and 10/0 °C conditions, the stress imposed on “Selva” surpassed that on “Toyonoka” with equivalent treatment durations. The short-day “Toyonoka” encountered stress of intensity greater than or equal to three levels after 9 days at 15/5 °C and 6 days at 10/0 °C, while the long-day “Selva” experienced stress of no less than three levels after 6 days at 15/5 °C and 3 days at 10/0 °C. The critical temperature for chilling injury in the photosynthetic system of short-day strawberry was lower than that of the long-day strawberry.In future endeavors, we envision expanding our investigations to encompass strawberry cultivars belonging to diverse photoperiod types. This expansion aims to unravel the physiological significance and inherent interconnections among distinctive spectral signatures. We also intend to delve into the ramifications of low-temperature stress during various developmental stages on strawberry yield and fruit quality, and continually enhance our understanding of the physiological and production-related impacts of cold stress on strawberries.


The objectives of this study were to: (i) review and provide a narrative synthesis of three-dimensional (3D) foot surface scanning methodological and statistical analysis protocols, and (ii) develop a set of recommendations for standardising the reporting of 3D foot scanning approaches.A systematic search of the SCOPUS, ProQuest, and Web of Science databases were conducted to identify papers reporting 3D foot scanning protocols and analysis techniques. To be included, studies were required to be published in English, have more than ten participants, and involve the use of static 3D surface scans of the foot. Papers were excluded if they reported two-dimensional footprints only, 3D scans that did not include the medial arch, dynamic scans, or derived foot data from a full body scan.The search yielded 78 relevant studies from 17 different countries. The available evidence showed a large variation in scanning protocols. The subcategories displaying the most variation included scanner specifications , scanning conditions , foot measurements and definitions used, and statistical analysis approaches. A 16-item checklist was developed to improve the consistency of reporting of future 3D scanning studies.3D foot scanning methodological and statistical analysis protocol consistency and reporting has been lacking in the literature to date. Improved reporting of the included subcategories could assist in data pooling and facilitate collaboration between researchers. As a result, larger sample sizes and diversification of population groups could be obtained to vastly improve the quantification of foot shape and inform the development of orthotic and footwear interventions and products. Human foot morphology is highly variable and is influenced by a broad array of factors, including age , sex 2]2], ethniThe impact of variation in foot shape and on footwear fit are significant barriers that consumers, clinicians, and industry face. With the use of three-dimensional (3D) scanning technology, detailed information about the foot can be obtained and analysed to quantify foot shape . This deAlthough 3D surface scanning has been used to measure foot shape since the mid-1990s , only reCapturing a 3D model of the foot has been achieved through a variety of methods since its inception. The type of 3D scanning systems have been varied, ranging from the use of stereophotogrammetry (which uses multiple photographs taken from different angles) to structured light (patterns projected on the object) and laser scanning systems (laser/s are repeatedly projected onto a surface while the camera/s and computer system acquires the 3D data) . Early sTo the best of our knowledge, with the exception of a broad overview of 3D foot scanning published in 2010 , and recThis scoping review was conducted and reported in accordance with the Joanna Briggs Institute methodology for scoping reviews  and the th, 2022, were included. Titles and abstracts were screened by two independent reviewers (JJA and HBM) for assessment against the inclusion/exclusion criteria for the review. Any disagreements on eligibility were resolved at a consensus meeting by a third independent reviewer (SEM).Three electronic databases were searched. An initial limited search of SCOPUS, ProQuest, and Web of Science were undertaken by two independent reviewers  to identify key studies for the topic. A reviewer (JJA) and research librarian (NP) identified key text words contained in the titles and abstracts of key studies and these were used to develop a full search strategy for SCOPUS, ProQuest, and Web of Science. Key search terms were grouped into three main concepts and adapted to each database: (i) 3D foot scanning, (ii) shape modelling/analysis/morphology, and (iii) foot/feet. Concept synonyms were searched under ‘topic’ which included title, abstract, and key words. Results from within each concept were combined with ‘OR’ and between concepts were combined with ‘AND’. The search strategy is provided in Supplementary file 1. Manual citation tracking and reference checking of included studies were performed. Grey literature such as conference proceedings were screened for additional studies. Only studies published in the English language were included. Studies published from inception to March 28To be included, papers needed to report studies that employed static 3D surface scanning of the foot. Studies with participants of any age, sex, geographical location, musculoskeletal pathology, or health setting were included. Papers were excluded if the studies used two-dimensional footprints only, 3D scans that did not include the medial arch, dynamic/sequential scans to infer foot movement, derived foot data from a full body scan, or diagnostic imaging techniques that may be used to create 3D models of the foot . Single case studies, studies involving 10 or fewer participants, non-English full text or conference proceedings/abstracts with less than four text pages were excluded due to a lack of detail provided, as were papers which described a scanning technique but provided no data on foot shape, and thesis dissertations. Foot dimensions measured were included if used in two or more studies.Study selection and data extraction were performed independently by two reviewers (JJA and HBM). Any disagreements on study eligibility were discussed between reviewers (JJA and HBM) and resolved at a consensus meeting by a third independent reviewer (SEM). A custom generated data extraction template was created in Covidence . The following individual study data were extracted from included studies: general study information , 3D scanner characteristics , study methods , 3D scanner methodology , scanner reliability (intra- and inter-rater), participant demographics , processing techniques , broad study design, statistical analysis approach, inclusion/exclusion criteria, and main outcomes. All 3D foot measures were cross checked using the IEEE and ISO definitions.A flowchart of included studies is shown in Fig. n = 41) [n = 6) [n = 4) [n = 3) [n = 3) [n = 3) [n = 3) [n = 3) [n = 2) [n = 2) [n = 2) [n = 2) [n = 2) [n = 2) [n = 1) [n = 1) [n = 1) [n = 1) [n = 1) [The included studies originated from 17 different countries: China n = 1 [4, 11,n = 41) –68, Japa [n = 6) , 69–72,  [n = 4) , 73–75,  [n = 3) , 77, Sou [n = 3) –80, Spai [n = 3) , 81, 82, [n = 3) –85, Ital [n = 3) , 21, 86, [n = 2) , Malaysi [n = 2) , New Zea [n = 2) , 62, Aus [n = 2) , 90, Bel [n = 2) , 91, Uni [n = 2) , 93, Can [n = 1) , Iran (n [n = 1) , Sloveni [n = 1) , and Swe [n = 1) , 93–95,  [n = 1) , 90, 14  [n = 1) , 90, 91, [n = 1) , 78, 81, [n = 1) , one com [n = 1) , one reg [n = 1) , and 11  [n = 1) , 84, 90, [n = 1) , and one [n = 1) . Thirty- [n = 1) , 94 and  [n = 1) –92, 94.n = 18) [n = 11) [n = 7) (such as Down syndrome [n = 8)  players [Sample size ranged from 11 to 1,200,847, with 57 studies including healthy young adults , 93, 94.n = 18) , 93, 94,[n = 11) , 89, 92,syndrome , 87, art players , America players , road cy players , collegi players , 71, rec players , 62, and players ), non-ha players , pregnan players , 47. Add players , 79, 86.2), and 23 reported standard deviation [All but six studies , 73, 79 eviation , 93, 94.eviation , 80, 88.Sixty-seven of the included studies performed 3D surface scanning of participants in bipedal stance  , 88–95, Seventy-two of the included studies reported the 3D scanner model. Twenty-five studies used the INFOOT USB scanning system  –89, 91, Twenty-seven studies reported 3D scanner intra-rater (test–retest) reliability , 90–93. A wide range of foot measures to quantify foot shape were reported within the included studies see Fig. . Sixty-fThirteen studies cited the ISO standards , 82, 88.Nineteen studies reported the processing techniques used. Sixteen studies used meshing , 94, 95,t-tests or ANOVAs) [r, Spearman’s rho, or intraclass correlation coefficients) [Forty-three studies reported analysing mean differences  , 91, 93,icients) , 87, 91,icients) , 87, 90.The purpose of this scoping review was to provide a description of the methodological and statistical analysis protocols used in 3D surface scanning of the foot. Overall, the included literature highlights large variability of methods used in 3D foot scanning. We identified four key areas that showed variability and paucity of reporting between 3D scanning methods, these included the wide range of scanners used , scanner condition protocols , statistical analysis techniques, and foot measures used. Additionally, the literature included a broad range of participants ; however, studies were heavily skewed to include healthy young adults of Chinese heritage. The predominance of young adults from Chinese heritage may be due to most studies being conducted in China and using a convenience sample. Furthermore, the included studies lacked data on pathological foot shapes . This underlines the need for a consistent approach in reporting methods for future 3D surface scanning research.The available evidence incorporated several 3D scanning models and associated scanning types to collect 3D foot shape data. The most common type of scanning system was laser scanning, with the INFOOT USB scanning system  being the most frequently used see Fig. . HoweverScanner reliability was scarcely reported, with only 21 of the 78 (26%) studies performing test–retest reliability and 13 (17%) reporting scanner calibration methods. This may be problematic as potential technical error of measurement of the observer conducting the scan could occur, and may not be identified without reliability and scanner calibration being performed prior to data collection . SimilarVariability in weightbearing condition definitions were evident among the included studies. The description for semi/partial weightbearing and non-weightbearing were inconsistent. The IEEE define non-weightbearing as “the foot is in the air when measured not supporting any bodyweight” . SeveralThe types of marker placement  were inconsistent between studies. Both methods have their limitations. The manual method is time consuming and relies on an experienced investigator to accurately palpate anatomical landmarks. Automatic marker placement is faster, although there may be errors during software estimation of landmark positions , 98. TheWhile all studies included in this review used 3D scanning technology and several authors applied PCA to determine foot shape, most studies performed PCA by extracting simple 1D or 2D measures  from the 3D foot scan , 42. ThiTwenty-two scanning measures were identified that satisfied our inclusion criteria of being reported in at least two studies. There was high variability in foot measure descriptor terminology and definitions across the included studies. This resulted in multiple descriptors being used for the same foot measurement. For example, ball width (a term used by the IEEE) has been described within the included studies as ‘ball breadth’, ‘forefoot width/breadth’, ‘linear width’, ‘ball section width/breadth’, ‘metatarsophalangeal joint width’, ‘stick width’, and ‘diagonal width’, and several descriptors have been used to identify foot length from the posterior aspect of the heel to the first metatarsal head, including ‘instep length’, ‘arch length’, ‘heel-to-ball-length’, ‘ball of foot length’, ‘medial ball reach’, and ‘medial arch length’. We also found that definitions of the same foot measure varied depending on the scanner model and definition of the foot axis used. The ISO has created standards for 3D body scanning; however, foot length and width are the only included foot measures . AdditioThis review has several strengths. Firstly, we conducted a rigorous search strategy with the assistance of a research librarian (NP) to ensure optimum coverage of the available literature. Secondly, two independent reviewers were involved in the title and abstract screening and data extraction, with any conflicts being resolved by a third reviewer. Thirdly, we followed the Joanna Briggs Institute methodology for scoping reviews and used a standardised template for the extraction, analysis, and presentation of results . HoweverConsistent ReportIng Three-dimensIonal sCanning) for the identified areas above to help improve protocol transparency and reporting consistency between studies , scanning conditions , and definitions and descriptions of scanner measurements. Additionally, the statistical analysis approaches used throughout the literature are highly variable with many studies utilising techniques that may not optimise the shape analysis capabilities of 3D scanning technology. Although international standards (ISO and IEEE) exist, they were not widely adopted in the included studies. Based on the findings of this review, we have developed a consistent reporting method (CRITIC 16 item checklist - The paper provides an overview of 3D foot scanning methodological and statistical analysis protocols. It can be concluded that inadequate reporting and lack of consistency in 3D foot scanning methodological and statistical analysis protocols exists. Despite the availability of international standards for 3D foot scanning, few studies utilised this resource and consequently there is a lack of homogeneity between scanning protocols. Additionally, the need for utilisation of optimal statistical analysis techniques of a diverse population group to quantify foot shape is needed. The improved consistency of 3D foot scanning methodologies and utilisation of more sophisticated statistical analysis techniques could enhance data pooling and collaboration between researchers. As a result, larger sample sizes, diversification of population groups, and simultaneous data collection could be obtained to enhance the quantification of foot shape and facilitate the development of improved orthotic and footwear products.