In several author name disambiguation studies, some ethnic name groups such as East Asian names are reported to be more difficult to disambiguate than others. This implies that disambiguation approaches might be improved if ethnic name groups are distinguished before disambiguation. We explore the potential of ethnic name partitioning by comparing performance of four machine learning algorithms trained and tested on the entire data or specifically on individual name groups. Results show that ethnicity‐based name partitioning can substantially improve disambiguation performance because the individual models are better suited for their respective name group. The improvements occur across all ethnic name groups with different magnitudes. Performance gains in predicting matched name pairs outweigh losses in predicting nonmatched pairs. Feature  similarities of name pairs vary across ethnic name groups. Such differences may enable the development of ethnicity‐specific feature weights to improve prediction for specific ethic name categories. These findings are observed for three labeled data with a natural distribution of problem sizes as well as one in which all ethnic name groups are controlled for the same sizes of ambiguous names. This study is expected to motive scholars to group author names based on ethnicity prior to disambiguation. One study estimates that about two‐thirds of author names in PubMed, the largest biomedicine digital library, are vulnerable to either or both of these two ambiguity types  methods. Some scholars have used heuristics such as string‐based matching , which is the most widely used approach in bibliometrics   are more difficult to disambiguate than others  dominates, this study disambiguates new labeled data in which all ENGs are controlled to have the same numbers of instances, to demonstrate that performance changes induced by ethnic name partitioning may not be solely due to the well‐known relationship between the number of cases and their ambiguity . Third, this study shows that different combinations of features  appear to be related to AND performance for different ENGs suggesting future directions to further improve AND performance with ambiguous ethnic names. The findings of this study can provide practical insights to researchers and practitioners who handle authority control in digital libraries. In the following sections, details on labeled data and setups for machine learning are described.This study aims to empirically evaluate the effect ethnic name partitioning has on AND. In this study, AND is a task to assign either “match” or “nonmatch” label to a pair of author name instances. For this, specifically, name instances are grouped into a block that share the same first forename initial and full surname and pairwisely compared within the block for their similarities over a set of features  to produce similarity scores. Machine learning algorithms combine the scores to learn weights of each feature to decide if a given pair of instances to refer to the same author or not. Although our work is motivated by the studies reviewed above and follows their common data preprocessing, blocking and machine learning steps, this paper differs from them in three important ways. First, this study evaluates AND performance by four different machine learning algorithms applied to four different labeled datasets before and after inclusion of a standard ENG partition. Here, a name instance is assigned to an ENG based on a name ethnicity classification system, 22.1To measure the effect of ethnic name partition on machine learning for AND, this study disambiguates names in four labeled datasets—KISTI, AMiner, GESIS, and UM‐IRIS. The first three datasets have been used in many AND studies to train and test machine learning algorithms  and Kyungsung University in Korea constructed this labeled dataset. It is made up of 41,673 author name instances that belong to 6,921 unique authors  in Germany produced this labeled data. It contains author name instances of 5,408 unique authors  through matching selected name instances in publication records to an authority database, ORCID , paper title, and publication venue—are used as machine learning features because they have been widely used in algorithmic AND studies Schulz,  and are 2.2This study assigns an ENG tag to a name instance in each labeled dataset using the author name ethnicity classification database, Ethnea, developed by Torvik and Agarwal .9 EthneaTable Chinese names represent the majority of ENGs in three labeled data. This is because these labeled data were created from computer science papers where Chinese researchers are particularly large contributors. In addition, as the three datasets were designed to collate challenging names to disambiguate, Chinese names that tend to be more ambiguous than other ENGs were oversampled  frequency for each feature  and test (50%) subsets. Then, instances in each subset are put into blocks in which all member instances share the same full surname and first forename initial . Only instances in the same block are compared for disambiguation. This blocking is typical in AND studies because it reduces computational complexity with only slight performance degradation  for a predefined set of ENGs.We focus on four algorithms—gradient boosting (GB), logistic regression (LR), naïve Bayes (NB), and random forest (RF)—for supervised machine learning that have been widely used as baselines or best performing methods in AND studies  and negative  pairs, respectively. In addition, we calculate the F1 score as a harmonic mean of precision and recall.Specifically, precision for positive pairs (PrecPos) measures how many predicted match pairs are correct ones (true positives [TP]) over the total number of predicted match pairs that may contain correct match pairs (TP) and incorrect match pairs . In contrast, recall for positive pairs (RecPos) measures the ratio of correct match pairs (TP) over the total number of true match pairs that may be predicted correctly as match pairs (TP) or incorrectly as nonmatch pairs ,Likewise, precision for negative pairs (PrecNeg) measures how many predicted nonmatch pairs are correct ones (true negatives [TN]) over the total number of predicted nonmatch pairs that may contain correct nonmatch pairs (TN) and incorrect nonmatch pairs (FN). In contrast, recall for negative pairs (RecNeg) measures the ratio of correct nonmatch pairs (TN) over the total number of true nonmatch pairs that may be predicted correctly as nonmatch pairs (TN) or incorrectly as match pairs (FP),The metrics are calculated on the entire set of test results for each labeled dataset. We separately calculate performance measures for different ENGs rather than averaging them across multiple ethnicity groups.33.1before and after ENG‐aware disambiguation by four algorithms—GB, LR, NB, and RF. Figure Figure Ethnic name partitioning also reduces the number of falsely predicted nonmatch cases (FN), increasing recall in Figure ENG‐aware disambiguation also does a better job of accurately predicting nonmatch (negative pair) cases. The “After” bars are taller than those of “Before” in Figure Algorithmic performances are also enhanced by ENG‐aware disambiguation on AMiner, GESIS, and UM‐IRIS. Figures 3.2ENG‐aware disambiguation produces substantial improvements in both precision and recall for predicting match and nonmatch instance pairs in different labeled datasets. But are those improvements uniform across different ENGs? If not, a more nuanced approach to model evaluation may be necessary. To answer this question, we compare performance changes due to ENG‐aware disambiguation within ENG groups. For this, precision, recall, and F1 scores for positive and negative pairs predicted by four algorithms are calculated separately for instance pairs that belong to the same ENG in each of four labeled data: 4 algorithms × 4 data = 16 evaluations. Presenting all the results at the same time would consume too much space in this paper. So, we present RF predictions on the GESIS dataset as an illustration for the purposes of this discussion. Reports of other algorithms and data are presented in Supporting Information attached to this paper.Figure x‐axis and the ratios of the numbers of scores in the same bin over all scores are plotted on y‐axis. A solid red line represents the distribution of all instance pairs regardless of ENG.These observations can be explained as follows. ENGs have different distributions of similarity scores over the four basic (nonethnicity) features we use. Figure coauthor” similarity scores for both positive and negative pairs  features, they do a better job of predicting nonmatch pairs based on aggregated feature similarity distributions that are invariant across particular ENGs. In other words, feature distributions aggregated across ENGs appear to be more effective for predicting negative case pairs while ENG‐aware disambiguation techniques more accurately capture positive pairs.These observations imply that disambiguation models for positive pair prediction would be improved by ENG‐aware procedures, while nonmatch patterns for negative pair prediction can aggregate across ENGs  in which Chinese names are dominant and the overall performance improvements were heavily affected by those for Chinese name instances. To overcome such an imbalance of instance distribution in labeled data, a new labeled data (UM‐IRIS) were created in a way that six ENGs have the same amount of ambiguous name instances. Disambiguation results from the new labeled data were in line with those from other three labeled data. In addition, all ENGs including Chinese were able to obtain gains in disambiguation performances. But all these findings were obtained from small‐sized labeled data, whether they are biased or controlled for ENG sizes. So, it is still unknown whether such improvements are achievable in AND for large‐scale bibliographic data in which ENG composition may be quite different from those in the labeled data used in this paper.Another issue would be that there can be other features than the four used in this study that can lead ethnic name partition to different AND performances. For example, English authors may appear in publication records that are more complete in affiliation information and use more diverse title terms. Meanwhile, Chinese authors may tend to work with coauthors who have similar names in same institutions. Various features need to be explored to study further the impact of ethnic name partition on AND.precpos↑, recpos↓), while “nonmatch” pairs of other ENG instances as matched ones . This might be because while some Chinese pairs sharing coauthor names, venue names, or title words refer to different authors, other ethnic names sharing the features are more likely to represent the same authors . As the sizes of negative pairs in most non‐Chinese ENGs are smaller than positive pairs  or negative (nonmatch) pairs.As detailed in the discussion above, ethnic name partition requires further research for us to better understand its impact on AND and apply it to disambiguation tasks for digital libraries that are struggling with authority control over fast‐growing ambiguous author names. This study is expected to motivate scholars and practitioners to study toward that direction by demonstrating the potential of ENG‐aware disambiguation in improving disambiguation performances.Appendix Supporting InformationClick here for additional data file.


While fdU is almost exclusively an oxidative stress lesion formed from deoxythymidine (T), the situation for fdC is more complex. Next to formation as an oxidative lesion, it is particularly abundant in stem cells, where it is more frequently formed in an epigenetically important oxidation reaction performed by α‐ketoglutarate dependent TET enzymes from 5‐methyl‐deoxycytidine (mdC). Recently, it was shown that genomic fdC and fdU can react with the ϵ‐aminogroups of nucleosomal lysines to give Schiff base adducts that covalently link nucleosomes to genomic DNA. Here, we show that fdU features a significantly higher reactivity towards lysine side chains compared with fdC. This result shows that depending on the amounts of fdC and fdU, oxidative stress may have a bigger impact on nucleosome binding than epigenetics. The covalent adduct formation of the oxidative lesions and epigenetic bases 5‐formyl‐deoxyuridine (fdU) and 5‐formyl‐deoxycytidine (fdC) was investigated comparatively arguing that oxidative stress can have transcriptional consequences. While g H‐bond,24, 251) containing a central lysine. This peptide 1 mimics a tryptic peptide with a charge distribution that is suitable for identification via MS. The amino group at the N‐terminus was protected with an acetyl group to block its reactivity towards formyl groups. In consequence, the peptide has only one amino group that can react. For the reaction, an excess of fdC or fdU was added to a solution of the peptide 1.In order to examine the reactivity of fdU towards lysine side chains in direct comparison to fdC, we prepared the acetylated heptapeptide AcNH‐IEAKGER‐OH . Although mass spectrometry is a priori not a quantitative method due to different ionization efficiencies of the analytes, the almost identical structures of the peptide adducts 4 and 5 allow to extract some quantitative information, assuming that the ionization properties of adducts 4 and 5 are comparable to each other. This assumption is based on previous studies which show that ionization efficiencies are mostly dependent on the molecular volume and the hydrophobicity of a species, which should both be very similar for the two adducts.[4 and 5 , we saw in both cases  no formation of any adduct .To investigate whether this reactivity difference persists when the fdU and fdC nucleosides are embedded in an oligonucleotide context, we prepared the DNA strands 1 Figure a. In thi9 on the denaturing gel  (lane h).Using this system, we now detected slight adduct formation for fdU (lane f), but clearly none for fdC (lane e) supporting a higher reactivity for fdU. Adduct formation in lane f is visible by the partial shift of fluorescence intensity to higher molecular weight bands for the fdU‐adduct l Figure b. Even bThese studies show that the reactivity of the two formyl‐containing bases is quite low in solution, which is expected because in water the equilibrium between Schiff base and aldehyde is far on the aldehyde side.11, 12) derived from a 145 bp region of the Widom 601 DNA sequence (10),11) or all Ts are exchanged against fdUs . For the PCR reaction of the modified strands, we had to carefully adjust various parameters in order to generate the 145mer containing either 54 fdUs or 69 fdCs. We achieved this by using the optimized polymerase blend KOD XL and lengthening the extension time (see Supporting Information), similar to what we described earlier.SYBR Green I. With this reagent it was impossible to visualize the nucleosome assembly with strand 12. We assume that this lack of detectability is caused by the structure of the heavily fdU‐modified DNA. The structural effects generated by the high density incorporation of fdU may prevent proper intercalation of the fluorophore into the duplex, as also a single fdU modification has been shown to affect DNA structure.Cy3 covalently attached to its 5’‐end in all PCR reactions. This generates the DNA duplexes 10–12 with a Cy3 dye label as the readout signal. Analysis of the nucleosome assembly by native PAGE using this Cy3 detection reveals that the fdU containing 601 sequence gives a slightly better assembly of the nucleosomes as compared to fdC. The amount of free DNA is clearly decreased in the assembly reaction with fdU compared to fdC and dC. We believe that this is already an effect of the higher reactivity of fdU towards the lysine amines.In order to compare the reactivity of fdC and fdU in the context of nucleosomes, we generated two longer dsDNA sequences  show only a band corresponding to free DNA, meaning that without formyl groups, no adducts can form. This changes for the formyl‐containing strands 11 and 12: even without reduction , we detected a few distinct bands at high molecular weight, which likely resemble stable fdC‐ and fdU‐DNA‐nucleosome complexes that do not denature on the gel. This is surprising, given the reversible nature of the non‐reduced Schiff bases. We postulate that these bands are likely the result of multiple adducts forming simultaneously at various positions of the histones, which collectively counteract the reversible nature of imine adduct formation. Indeed, when we heated the non‐reduced complex to 95 °C for 5 min in sample buffer before loading onto the gel , these bands mostly disappeared supporting this interpretation.To investigate the reactivity of fdC and fdU, we allowed the reconstituted nucleosomes to react for 24 h at 37 °C. In order to stabilize the Schiff bases, we next added NaBH3CN  yields similar bands compared to those observed with the non‐reduced samples, although for fdU there are more diffuse bands at higher molecular weight, which could be attributed to reductively stabilized adducts with different stoichiometry than before. Because of reduction, these adducts are now mostly resistant to dissociation by heating . Finally, we treated these complexes with proteinase K to fully digest the histone proteins . In this case, we see that the DNA‐nucleosome adducts disappear, leaving just a diffuse DNA‐only band behind. This confirms the involvement of the histone proteins in complex formation. Judging from the sharper band obtained from the released fdC‐containing DNA 11 (lane g) compared to the fdU‐containing DNA 12 (lane l), one can conclude that the digestion of the fdC‐complex is more complete. This further strengthens the idea that the fdU‐containing complexes are more stable and hence more compact, which leads to reduced accessibility for proteolysis.Informative, however, is a comparison of the products that result from heating between the fdC and fdU sample. For fdC (lane d) mainly the intensity of the band representing free DNA increases, arguing that the nucleosomal complex dissociates completely. In contrast, preheating of the fdU nucleosome (lane i) leads to many additional diffuse bands in addition to an increased intensity of the free DNA band. This suggests some complete but also partial denaturation, as well as aggregation of the nucleosomes in the case of fdU. These combined results argue that the fdC adducts are less stable, which provides a higher reversibility. Addition of NaBH601 context. Despite this, we see that it creates crosslinks more readily, arguing that we may even underestimate the reactivity differences. Since it was already shown that fdC and fdU can change the local structure of DNA,[601 sequence. For this, we recorded CD spectra of the DNA strands 10–12 in the same buffer we used for nucleosome reconstitution . This revealed that while the unmodified strand 10 is present as B‐DNA, the fdC‐containing strand 11 adopts a different conformation. This conformation features similar CD properties to what has been reported for multiple fdC modifications contained in shorter DNA, termed F‐DNA.12 exhibits a CD spectrum that resembles A‐DNA.Our experiments were performed with artificial nucleosomes, in which fdU and fdC occupy different positions within the complex . This could obscure a direct reactivity comparison. It should however be noted that fdU occupies fewer sites on the nucleosome in the e of DNA,34, 351 and fdC and fdU nucleoside or short fdC‐ and fdU‐containing ssDNA. It seems that the simultaneous formation of multiple Schiff bases is beneficial to stabilize the adducts. This would mean that predominantly clustered fdC as suggested to occur by Balasubramanian and co‐workersIn summary, we confirm the previously reported Schiff base adduct formation between fdC‐ and fdU‐containing DNA and histones.16, 17The fact that fdU forms Schiff base adducts so efficiently could establish an additional mechanism of how oxidative stress and clustered oxidative lesions in particular can induce epigenetic processesBased on previous reports, it is furthermore likely that Schiff base formation affects the accessibility of fdU and fdC,19, 20The authors declare no conflict of interest.As a service to our authors and readers, this journal provides supporting information supplied by the authors. Such materials are peer reviewed and may be re‐organized for online delivery, but are not copy‐edited or typeset. Technical support issues arising from supporting information (other than missing files) should be addressed to the authors.Supporting InformationClick here for additional data file.


Complex data processing and curation for artificial intelligence applications rely on high‐quality data sets for training and analysis. Manually reviewing images and their associated annotations is a very laborious task and existing quality control tools for data review are generally limited to raw images only. The purpose of this work was to develop an imaging informatics dashboard for the easy and fast review of processed magnetic resonance (MR) imaging data sets; we demonstrated its ability in a large‐scale data review.We developed a custom R Shiny dashboard that displays key static snapshots of each imaging study and its annotations. A graphical interface allows the structured entry of review data and download of tabulated review results. We evaluated the dashboard using two large data sets: 1380 processed MR imaging studies from our institution and 285 studies from the 2018 MICCAI Brain Tumor Segmentation Challenge (BraTS).3) than did segmentations with minor errors (20.5 cm3) or failed segmentations (27.4 cm3). In the BraTS data, 242 of 285 (85%) studies were acceptable quality after processing. Among the 43 cases that failed review, 14 had unacceptable raw image quality.Studies were reviewed at an average rate of 100/h using the dashboard, 10 times faster than using existing data viewers. For data from our institution, 1181 of the 1380 (86%) studies were of acceptable quality. The most commonly identified failure modes were tumor segmentation (9.6% of cases) and image registration (4.6% of cases). Tumor segmentation without visible errors on the dashboard had much better agreement with reference tumor volume measurements (root‐mean‐square error 12.2 cmhttps://github.com/EGates1/MRDQED.Our dashboard provides a fast, effective tool for reviewing complex processed MR imaging data sets. It is freely available for download at  But, they focus almost exclusively on the quality and statistics of the raw images themselves. They do not help with the review of other facets of the data, such as registrations or segmentations, which are critical to quantitative measurements. Thus, there is a need for a single tool for both systematically reviewing image quality and checking the accuracy of the derived images and measurements. The goal of this project was to develop a review interface for MR data sets that had several key characteristics:Our solution was to develop a dashboard to quickly and efficiently visualize all of the necessary image data for a single case and record the quality of the various images, masks, etc. that affect the final image measurements. We rendered a handful of key slices as static portable network graphics (PNG) image files as part of the data processing pipeline: essentially, we front‐loaded the cost to load the data at the expense of the ability to freely scroll through image slices. Reviewing just a few orthogonal slices through segmentation or image was generally sufficient to screen for unacceptable data quality.We used the dashboard to efficiently review 1380 brain tumor imaging studies that were processed as part of a large‐scale research study. In addition to calculating the various failure rates in our data‐processing pipeline, we identified which studies had acceptable data quality, which contained minor errors, and which should be excluded from further analysis. Using this classification, we compared the segmented tumor volume with the reference measurements to show how higher quality segmentations have smaller average errors. We also tested our dashboard using 2018 MICCAI Brain Tumor Segmentation Challenge (BraTS) data (285 studies).2https://github.com/EGates1/MRDQED. The dashboard code is flexible enough to use for most projects involving 3D image datasets with annotations saved in NIfTI format.We implemented a dashboard interface that systematically displays representative slices of each image and segmentation using the R Shiny package.The app loads a data file with case IDs, image file paths, and any other desired tabular data. Before the app runs, the images and segmentation are used to generate key PNG snapshots. We rendered PNGs of axial, sagittal, and coronal slices for each image, with each segmentation overlaid as part of our existing data processing pipeline. The slice displayed was the one with the maximum area in the segmentation. As part of this process, the various segmentations  were loaded together and assigned separate distinct colors. We also rendered scaled density plots of the combined segmentation, as well as three‐plane crosshair views centered on the maximum intensity voxel within the tumor segmentation for each image. By rendering snapshots as part of the initial data processing (and not during data review), the computational time is effectively front‐loaded so that all the dashboard does is render the existing PNGs.Study artifact: All images in the study have large artifacts, are corrupted or are otherwise not fit for further data processing.Image unusable: One or more images are not fit for further data processing.Bias field: Either a strong visible bias field is visible on one or more images or bias correction was applied that has corrupted the intensities.Normalization failure: The intensity range on the image histograms is unreasonably outside of the expected range.No PNGs: Some PNGs are missing or not displayed in the app.Needs review: Closer inspection of the study is necessary outside of the dashboard.A screenshot of the app is shown in Figure Below the checkboxes is a free text comment box to add additional review notes. After reviewing a case, the “save + next” button loads the next case to review in 3–5 s. There is also a download button that exports a comma‐separated values file of the reviews entered via the current web browser. R Shiny supports multiple concurrent user sessions but does not save data between sessions. Thus, each time a reviewer connects, they can upload the latest review log to re‐populate the fields and continue.The top left of the data display panel Figure  displaysWe evaluated the dashboard using two large data sets. Each data set was consecutively reviewed by a single reviewer over several reading sessions. We compiled the results of the review to discover the most common failure modes and what proportion of the processed data was acceptable.First, we used a historic data set of 1380 glioma patients from a single institution. Preoperative brain MR imaging (MRI) studies were downloaded from the picture archiving and communication system and processed, including co‐registration,3The review of all 1665 studies was completed by a single reader (5 years of experience) at an average pace of 100 studies/h. In the authors’ experience, reviewing the same data manually using standard open‐source data viewers (Insight toolkit ITK‐SNAPn = 29) or raw image quality (n = 14). These 14 cases primarily had excessively cropped fields of view that overlapped with ground truth tumor segmentations. Some examples of this are shown in Figure The resulting review logs were used to categorize each study as acceptable, acceptable with minor errors, or unacceptable because of poor data quality  or data processing failures. The numbers of cases of each quality are listed in Table 3 which was 55% smaller than the average error for failed segmentations (27.4 cm3) and 40% smaller than the average error in segmentations with minor inaccuracies (20.5 cm3). Failed segmentations had 276% higher average error in enhancing volume than segmentations with acceptable quality or minor errors (25.7 cm3 vs. 9.3 cm3). Figure As an example of the benefit of data review, we calculated the error in segmented tumor volume for each level of data quality  on the data from our institution using reference tumor volumes from previous clinical research studies. For this, we only used cases that had acceptable image quality, registrations, and brain masks so that the tumor segmentation was the deciding factor in the data quality. As expected, better quality segmentations had a smaller average error in tumor volume. This was the case for both total tumor volume (T2‐FLAIR hyperintensity) and T1‐enhancing volume (Table 4We implemented a lightweight, efficient review dashboard for MRI data sets. The dashboard enabled a complete, structured, human review of data quality of 1380 studies from our institution and 285 publicly available studies. The data review procedure provided useful information on the failure rates in various parts of our data processing pipeline and uncovered possibly problematic images in the BraTS challenge data.There are published tools for graphically displaying imaging data with the goal of assessing quality.Dashboards are not the only way to identify unwanted outliers in imaging data sets. Deep‐learning‐based segmentation models can provide their own uncertainty measurements, which can help identify low‐quality data.In addition to its many benefits, the dashboard approach has a few limitations, primarily related to the trade‐off between review speed and completeness. We chose fixed, pre‐rendered two‐dimensional PNG images that displayed key informative slices for each image. This reduced amount of data takes less time for a human to review while still enabling quality assessment. The limitation is that artifacts or segmentation errors may not be visible on those key slices. However, we felt that our desired level of data review—acceptable, minor errors, or failure—did not require a substantial increase in review time and complexity. The other limitation is that the imaging data cannot be edited inside the dashboard and any small errors must be handled in separate software. However, we found that existing tools for editing segmentations by hand were far superior.5We developed a lightweight, efficient, easy‐to‐use data review tool to evaluate imaging data sets. It allows the confident evaluation of the quality of a very large amount of data in a reasonable amount of time. Through our review, we characterized the failure rates in our data processing pipeline and found that higher data quality overall was correlated with smaller volumetric errors. A simplified version of our dashboard that can be used for other MR research data sets is available online.Evan Gates, David Fuentes, and Dawid Schellingerhout conceptualized the project. Evan Gates and Adrian Celaya tested and evaluated the dashboard. Evan Gates collected the final data and analyzed it. Dawid Schellingerhout and Dima Suki provided clinical expertise and supervision for the project. Evan Gates, Adrian Celaya, and David Fuentes drafted the manuscript. All authors reviewed approve of the final manuscript.


Suaahara (2011–2023)—aims to improve nutrition behaviours. Suaahara II (2016–2023) transitioned from a mother/child dyad focus to explicitly targeting all family members. Evidence is scant, however, regarding how exposure by men to social and behaviour change interventions relates to nutrition outcomes. This study uses a 2019 cross‐sectional monitoring dataset to test associations between maternal and male household head exposure to Suaahara II interventions  and adoption of three infant and young child feeding practices: minimum dietary diversity, minimum acceptable diet and sick child feeding, in households with a child under 2 years (n = 1827). Maternal exposure to Suaahara II had a positive association with minimum dietary diversity , minimum acceptable diet  and increased feeding to a sick child . Male household head exposure was only associated with increased feeding to a sick child . Among households with an exposed mother, having an exposed male household head nearly tripled the odds of appropriate sick child feeding  but was not significantly associated with the other two outcomes. These findings suggest that the relationships between exposure to nutrition programmes and outcomes are complex and further research is needed to understand variation by family member, behavioural outcome and context.In Nepal, an at‐scale, multisectoral programme— Approximately three‐fourths of mothers and more than one‐third of male household heads were exposed to a multisectoral nutrition programme 2 years after the programme started.Suaahara II and minimum dietary diversity, minimum acceptable diet and sick child feeding were found.Positive associations between maternal exposure to Suaahara II interventions was associated with an increased odds of 2.21 for appropriate sick child feeding and nearly tripled the odds of this ideal practice in households with an exposed mother, but no association was found between male household head exposure and either minimum dietary diversity or minimum acceptable diet.Male household head exposure to These findings suggest that the family approach may be effective at increasing adoption of some ideal complementary feeding practices but that more research is needed to understand how engaging male family members relates to adoption of ideal child nutrition practices in different contexts.11.1It is widely agreed that the thousand‐day period encompassing the 9 months of pregnancy plus the first 2 years of an infant's life is a window of opportunity for interventions, precisely because nutritional well‐being at this critical time will impact the child for the rest of his or her life  outcomes in low‐ and middle‐income countries is a high priority of governments and development partners alike  USAID‐funded multisectoral nutrition programme that supports Nepal's MSNP, builds off of the first 5‐year phase of the programme (2011–2016) and operates in 42 districts of Nepal to reach over 1 million households in the 1000‐day period ; homestead food production and agricultural marketing; and nutrition governance  and monitoring, evaluation and research (MER) for learning. Suaahara II works at national and subnational levels to improve nutrition policies and stakeholder coordination and supports a cadre of Government of Nepal health, agriculture and WASH frontline workers to improve quality and inclusive services. Suaahara II household and community programming primarily operates via social and behaviour change communication platforms including interpersonal communication, community mobilization, mass media and more recently SMS messages and social media. Suaahara II hired and trained field staff to conduct home visits and facilitate community events such as food and handwashing demonstrations, usually done during health mothers' group meetings, in collaboration with Nepal's Female Community Health Volunteers. The mass media edutainment radio programme, titled Bhanchhin Aama (Mother Knows Best), covers the wide range of practices that Suaahara II seeks to influence. These interventions target mothers and family members in 1000‐day households to reinforce key messages and promote optimal nutrition‐related practices  and developed a toolkit to provide interpersonal communication to all family members instead of only mothers and to encourage the whole family to participate in community events. Bhanchhin Aama (Mother Knows Best) was also revised in July 2017 to include new characters, namely, an adolescent girl and grandparents, to expand the programme's appeal to the whole family.In the fall of 2016, Suaahara II by both mothers and male heads of household and adoption of ideal infant and young child feeding (IYCF) practices in households with a child under 2 years after roll‐out of the family approach. This study further explores the association between male household head exposure and these key outcomes limited to households where the mother is exposed to test the theory that two exposed adults, versus one, improves the odds of adopting the ideal behaviours.To date, however, the benefits and risks of the transition to this family approach vis‐à‐vis MCHN behaviours that the programme aims to improve are unknown. There is scant evidence on family approaches to nutrition programming, especially those engaging multiple adult family members on multiple MCHN outcomes. This manuscript starts to fill these gaps by testing associations between exposure to 22.1Suaahara II. The general objective of this cross‐sectional monitoring survey is to monitor the progress of key maternal and child health and nutrition indicators over time. The data were collected from June to September in 2019 across all three agro‐ecological zones—mountains, hills and terai (plains)—and are representative of the 42 of Nepal's 77 districts in which Suaahara II interventions are implemented. Multistage cluster sampling was used to randomly select 16 districts; 32 municipalities ; 96 wards ; 192 subwards (two per ward); and 3648 households with a child under 5 years (19 per cluster). Probability proportional to size (PPS) sampling was employed to select study areas, followed by a listing of all households with a child under the age of 5 with the child's mother in residence and 19 of these eligible households per cluster randomly drawn from a bowl. In each selected household, the youngest child under 5 years was selected as the child for the study.This paper used a cross‐sectional monitoring dataset for an at‐scale, integrated programme in Nepal, known as Suaahara II exposure.The primary survey respondents were the mother of the selected child and the head of household, male if available. The mothers' questionnaire asked questions on practices related to MCHN topics such as antenatal care, delivery and postnatal care, childcare, and IYCF, as well as general health seeking, agriculture and homestead food production, and WASH. The household heads' questionnaire asked questions on household demographics and economics, household food security, land use and agricultural practices, and included observations on the household structure. Both surveys included a 24‐h dietary recall and asked about various domains of household‐level empowerment, integrated nutrition knowledge and exposure, self‐efficacy, gender‐based violence and psychosocial well‐being, and 2.2Suaahara II interventions: (1) met with a Suaahara II FLW in the past 6 months (yes/no); (2) ever participated in a Suaahara II community event such as a food demonstration (yes/no); and (3) ever listened to radio programme Bhanchhin Aama (yes/no). Given low levels of exposure to each type of intervention, a final summary binary variable was created to measure exposure to any of these three intervention types  developed an indicator for child minimum dietary diversity to measure the proportion of children between 6 and 23 months of age who consume foods from 4 or more of 7 food groups  over a 24‐hour dietary recall period , meaning that the child had at least the minimum dietary diversity and the minimum meal frequency during the previous day ; the agro‐ecological zone of residency ; whether the household was a member of a socially excluded caste or ethnic group defined as Dalit, Muslim or disadvantaged Janajati (yes/no); whether the household reported food insecurity (yes/no); and the socioeconomic status of the household . Only about half of these households had a male household head (n = 942) respond to the survey. Therefore, the sample sizes vary for each model, depending on the exposure variable (which household member) as well as the outcome variable. We conducted reverse power calculations to estimate the minimum detectable effect size for each of these models , to estimate associations between Suaahara II exposure by the male household head and all outcomes.Logistic regression models were first run to estimate associations between exposure by mothers and male household heads and outcomes of interest separately. We then used logistic regression models, limited to households where the mother was exposed to at least one of the three All analyses were adjusted for the individual and household level potential confounding factors mentioned above. All models were also adjusted for clustering. All statistical analyses were performed using Stata version 16.1 (2020).2.4Ethics approval was obtained from the Nepal Health Research Council (NHRC) in 2019. Respondents gave written informed consent to participate in the survey and reaffirmed their consent to continue their survey after completing each module in the questionnaires before continuing to the next module.33.1Household characteristics and key outcomes among sample households overall, as well as disaggregated between those with and without a male household head, are presented in Table P < 0.001), were from higher equity quintiles on average , and more resided in extended families . Mothers in the households with a male household member in residence were slightly more educated . Children were slightly younger in households with a male household head (11 vs. 12 months), but neither child wellness nor IYCF outcomes differed overall in these two types of households (P < 0.001).Households with a male household head in residence, compared to households without a male household head, were found to have a higher prevalence of radio ownership  reported an interaction with a frontline worker in the past 6 months and only 4% had ever participated in a community event. The bulk of exposure was from ever having listened to Bhanchhin Aama, with nearly one‐third (31%) of male household heads reporting having listened.On the other hand, over one‐third of male household heads (37%) had been exposed to at least one of the three types of 3.3Suaahara II and three key IYCF outcomes.Table Suaahara II was positively associated with all three of the child nutrition outcomes: minimum dietary diversity , minimum acceptable diet , and increased feeding to the child when s/he is sick . Male household head exposure to Suaahara II, however, was only associated with one of the three child nutrition outcomes: increased feeding to the child when s/he is sick .In the adjusted regression models, we found maternal exposure to Suaahara II and the three IYCF outcomes, only in households where the mother was exposed to enable testing to compare whether exposure by more than one adult family member (versus one) increases the odds of adoption of the promoted practices. Among households where the mother was exposed to Suaahara II, in final adjusted models, we found a positive, significant association between male household head exposure and a nearly three‐fold increase in feeding the child more food when s/he is sick . We found no significant associations between male household exposure, in households where mothers have been exposed, and the other two IYCF outcomes.Table 4Suaahara II frontline worker in the past 6 months, participation in a community event such as a health mothers' group meeting or a food demonstration, or listening to the Suaahara II radio programme, known as Bhanchhin Aama. Positive associations were found between maternal exposure to Suaahara II and all three outcomes: minimum dietary diversity, minimum acceptable diet, and increased child feeding when the child is sick. Male head of household exposure to Suaahara II was associated with increased feeding to a sick child, but not the other two outcomes. Furthermore, in models limited to households where the mother was exposed to Suaahara II, the addition of male household exposure was positively associated with increased feeding to the child during illness.This study estimates associations between different adult family members' exposure to multi‐sectoral nutrition interventions, controlling for socio‐economic and demographic characteristics, and household adoption of promoted maternal and child nutrition behaviours, among households in the 1000‐day period in Nepal. Two years after the start of the intervention, after roll‐out of the family approach, three‐fourths of mothers and over one‐third of male household heads were exposed to at least one of three types of interventions: interactions with a Suaahara II was much greater than that of male household heads, which is consistent with the fact that programmatic interventions during the first phase of Suaahara and beginnings of Suaahara II were focused on mothers while the family approach was being designed and formative research conducted. The initial interventions may not have been perceived by fathers and other male adults in the household as relevant and worth the effort to engage. Similarly,  frontline workers may not have approached males as often either for interpersonal communication or to encourage community event participation; despite training and the explicit transition to a family approach, this seems to have remained the case. Specific programme activities, such as the men only community groups shown effective in Bangladesh recently, may be necessary to generate engagement among male household heads  Cooperative Agreement AID‐367‐A‐16‐00006, provided financial support for this survey and analysis. This study is made possible by the generous support of the American people through USAID. The contents are the responsibility of the authors and do not necessarily reflect the views of USAID or the United States Government.KC and DN designed the study. RA, KC, PG and DN participated in data cleaning and statistical analyses. KC and DN prepared the first draft of the manuscript. RA, KC, PG, DN and SS provided revisions and additional text to manuscript drafts. KC and DN prepared the final manuscript, and all authors read and approved the final version for submission.The authors declare that they have no competing interests.Table S1: Infant and young child feeding outcomes among sampled households and minimum detectable effect size, 2019Click here for additional data file.


Adolescent girls and young women (AGYW) are a priority population for pre‐exposure prophylaxis (PrEP), a highly effective HIV prevention method. However, effective PrEP use among AGYW has been low. Interventions to support PrEP effective use may improve pill‐taking. Affordability of PrEP programs depends on their cost. We, therefore, evaluated the cost of community‐based PrEP with effective use counselling.Cost data from a randomized controlled trial were used to evaluate the cost of PrEP provision with effective use counselling offered to AGYW through community‐based HIV testing platforms between November 2018 and November 2019. AGYW were randomized to receive (1) group‐based community health club effective use counselling, (2) individualized effective use counselling or (3) community‐based PrEP dispensary. Task shifting of effective use counselling from nurses to trained lay counsellors was implemented in groups 1 and 2. Personnel costs were estimated from time‐and‐motion observations and staff interviews. Expenditure and ingredients‐based approaches were used to estimate costs for medical and non‐medical supplies.In total, 603 AGYW initiated PrEP and accrued a total of 1280 months on PrEP. Average cost per person‐month on PrEP with group‐based community health club, individualized effective use counselling and community‐based PrEP dispensary under the Department of Health scenario were similar and high  due to low PrEP client volume observed in the clinical trial. Increasing client volume  reduced cost per‐person month estimates to USD $15.48, $26.40 and $13.99, respectively.As designed, individualized effective use counselling increased the cost of standard‐of‐care PrEP delivery by 89%, group‐based community health effective use counselling increased the cost of standard‐of‐care PrEP delivery by 11%. These estimates can inform cost‐effectiveness and budget impact analysis for PrEP provision with effective use counselling services. Research studies for differentiated PrEP service delivery models outside sub‐Saharan Africa have shown that delivery of PrEP in community pharmacy settings  and teleWe, therefore, evaluated the incremental cost of community‐based PrEP provision and effective use counselling for AGYW using data collected from a randomized, controlled clinical trial.22.1Community‐based HIV counselling and testing (CBCT) offers HIV prevention, counselling and testing services at locations outside clinic settings. CBCT centres have the benefit of being located near where people live, taking HIV testing services outside of the traditional clinical setting  and offering youth‐friendly services. These benefits increase the yield of linking both HIV‐negative and HIV‐positive individuals to prevention and treatment services . The ComWe will refer to the three arms as standard care, clubs and individual, respectively. Medication dispensary was done by the nurse for all participants. Trained social workers provided counselling for arms 2 and 3. PrEP initiation procedures were the same for all participants, regardless of randomization. All PrEP service delivery in the study was provided within an existing CBCT platform by CBCT HIV care staff and social workers who received training specifically for this study. Enrolled AGYW were asked to return for an orientation visit 2 weeks post‐enrolment during which test results at enrolment were reviewed and more information about PrEP uptake was shared. Thereafter, monthly follow‐up visits were scheduled for 12 months, with all follow‐up visits occurring at the same site where participants enrolled. HIV testing was done every 3 months for the duration of the study period.https://clinicaltrials.gov/ct2/show/NCT03977181.Prior to enrolment, all participants provided written informed consent, which included counselling about randomization, procedures in each study group and their rights as research participants. The University of Cape Town Human Research Ethics Committee approved a waiver for guardian proxy consent for minors who opted not to involve parents via the Ministerial Consent for Non‐Therapeutic Research with Minors. The study protocol  was revi2.2Cost data for PrEP delivery were collected for programmatic activities, which included (1) start‐up activities: capital inputs , demand creation (posters and flyers) and initial PrEP training; and (2) ongoing activities: quarterly refresher trainings, supervision and administration , overhead costs  and direct service delivery activities .Study budgets and expense reports were utilized to estimate staff salaries for all personnel. Total and unit costs of recurrent components, which include clinical supplies, utilities, rentals, laboratory tests, medical and non‐medical supplies, were estimated using invoices, receipts, claim forms and contract agreement forms in case of services offered by contractors. Effective use counselling service costs and personnel time  costs were estimated using data collected through time‐and‐motion observations and clinical staff interviews. Cost data for demand generation activities – posters, flyers and household visits were used to estimate the cost of getting one AGYW to be screened for PrEP provision at CBCT platforms. Costs were collected in South African Rand (ZAR) and converted to United States dollars (1 USD = 14.31 ZAR) using an average monthly market exchange rate calculated for the data collection period from 1 November 2018 to 1 November 2019 .2.3We estimated the average cost per person‐month of PrEP medications dispensed for each service delivery model from a provider perspective following the Global Health Cost Consortium Reference Case (GHCC)  and PrEPCosts are either fixed or variable Table . Fixed cWe computed personnel time at PrEP initiation visits, follow‐up visits and effective use counselling sessions for each service delivery model by observing the total time needed and removing both waiting time for participants and time dedicated to research activities. Visits were classified as monthly refill visits  and quarterly refill visits . with all follow‐up visits occurring at the same site where participants enrolled. We averaged observed times to estimate personnel time spent on PrEP delivery activities. Clinical personnel cost was estimated by multiplying the observed average time spent on each PrEP delivery activity by the personnel cost per minute . Overhead costs for PrEP provision through CBCT platforms were proportionally allocated according to the fraction of total activities  that were dedicated to PrEP provision. PrEP medication costs per month for the as‐implemented scenario include the commodity cost as well as importation and central storage and distribution fees. DoH PrEP medication cost per month was obtained from recently published data  . We assu3A total of 603 AGYW initiated PrEP, accruing 1280 total months  of PrEP between 1 November 2018 and 1 November 2019 . The maximum number of clients who can be served per day is, therefore, constrained by the health worker with the total maximum time for the service  offered. For example, in standard care service delivery model, we use nurses’ time  to determine the maximum number of clients who can be served per day.The number of clients who can be served in a day for each service delivery model is shown in Table 3.1PrEP initiation visits required 51 minutes  of staff time, with clinical consultation accounting for approximately 63% of the total time, while the remaining time was spent with social workers. Scheduling calls and sending a text message for effective use counselling and medication pick‐up reminders to participants required 2 minutes per participant per visit  of social workers’ time.A median of 6 minutes  was required for a monthly standard care refill visit and 17 minutes  for a quarterly standard care visit. In both standard care visits, less than one tenth of the time was spent with social workers. The time required for club monthly visits had a median of 43 minutes , and 57 minutes  were required for quarterly visits. There was considerable variation in the duration of monthly refill visits because the duration of effective use counselling sessions depended on the extent to which young women spoke and interacted during the session. Monthly individual refill visits were a median of 30 minutes , with most services  provided by a social worker. Individual quarterly visits required 46 minutes , with 56% of the services provided by social workers.3.2The total annual PrEP delivery cost in the as‐implemented scenario was similar across the three service delivery models  , given the lack of staffing time for effective use counselling in the standard care service delivery model and having group effective use counselling sessions for the club service delivery model. The main driver of cost in the standard care service delivery model is PrEP medication (34%), followed by fixed personnel costs (29%), with a similar pattern for the club service delivery model . A flip in the main drivers of cost was observed in the individual service delivery model, with fixed personnel costs (31%) accounting for the largest proportion of costs, followed by PrEP medication costs at 18%.4In this micro‐costing study, we estimated that the cost to DoH of at‐scale PrEP provision integrated with group‐based community health clubs effective use counselling for AGYW resulted in a minimal increase of 11% compared to the cost of standard‐of care PrEP provision, while providing individualized effective use counselling resulted in an increase of 89%. PrEP medication accounted for the largest proportion of the cost per person‐month of PrEP, at 31%, consistent with other PrEP costing studies . Under tThe low volume of participants also resulted in similar estimates of the cost per‐person month of PrEP in the as‐implemented scenario and DoH scenarios. Conversely, in the scaled DoH scenario, we assume that PrEP uptake will increase as community knowledge of PrEP expands, and the concomitant increase in PrEP client volume substantially reduces the cost per person‐month estimates. Critically, evidence on the effectiveness of these strategies is needed to assess cost‐effectiveness and to guide the choice between these approaches. The costly service delivery models can be designed differently to cost less while at the same time maintaining their effectiveness if they are found not to be cost‐effective.The cost per person‐month estimates presented here were obtained from a study carried out in existing CBCT centres with trained and dedicated staff to perform effective use counselling and collect required samples for clinical tests. While CBCT centres do not exist in all parts of South Africa , these rAmong the DoH‐prescribed tests, HIV serology ($4.44) and HBsAg ($10.12) tests were the most expensive. These costs highlight the need for negotiated reduced costs. For example, HIV self‐test kits are now relatively low cost ($2) as a result of a buydown agreement negotiated by the Gates Foundation . The newWe excluded etiologic sexually transmitted infection (STI) testing from our cost estimates since it is not part of the standard‐of‐care of PrEP provision in South Africa. However, results from PrEP demonstration projects have reported high prevalence of STIs among young African women , 28, theThe lack of data describing the cost of real‐world PrEP implementation programmes has contributed to wide variation in the results of PrEP budget impact and cost‐effectiveness analyses , 13. Mey5Individualized effective use counselling increased the cost of standard‐of‐care PrEP delivery by 89%, while group‐based community health effective use counselling clubs increased the cost of standard‐of‐care PrEP delivery by 11%. These estimates may be useful to policy makers in planning PrEP implementation and can inform cost‐effectiveness and budget impact analysis for PrEP provision with effective use counselling services in CBCT platforms.This publication was made possible through a grant from the National Institute for Mental Health (NIMH), U.S. National Institutes of Health (NIH) under award number R01MH114648 to Professors AMM and LGB. The funder had no role in the design of the study and the collection and analysis of the data and in writing the manuscript.RVB has received conference abstract support from Regeneron Pharmaceuticals outside the submitted work. For the remaining authors, none were declared.EM led conception of the manuscript, design as well as drafting the manuscript, data analysis and interpretation of results with contributions from KP and RVB. JS and SM contributed to the acquisition of the data, analysis, and interpretation of the data and critical revision of the manuscript. LGB, AMM and CB contributed in the conception and design and critical revision of the manuscript. All authors reviewed and approved the final version of the manuscript.Data S1: Data – cost calculations spreadsheetClick here for additional data file.


Volatile organic compounds are important mediators of mutualistic interactions between plants and their physical and biological surroundings. Volatiles rapidly indicate competition or potential threat before these can take place, and they regulate and coordinate adaptation responses in neighbouring plants, fine‐tuning them to match the exact stress encountered. Ecological specificity and context‐dependency of plant–plant communication mediated by volatiles represent important factors that determine plant performance in specific environments. In this review, we synthesise the recent progress made in understanding the role of plant volatiles as mediators of plant interactions at the individual and community levels, highlighting the complexity of the plant receiver response to diverse volatile cues and signals and addressing how specific responses shape plant growth and survival. Finally, we outline the knowledge gaps and provide directions for future research. The complex dialogue between the emitter and receiver based on either volatile cues or signals determines the outcome of information exchange, which shapes the communication pattern between individuals at the community level and determines their ecological implications at other trophic levels. Released volatiles provide information about the identity and physiological status of neighbouring plants. The unique volatile profiles and release of specific cues and signals are guiding plants to response in a timely manner to cope with upcoming challenging threat, stress and to adapt to neighbours. Consequently, plants have adapted a variety of strategies to rapidly exchange volatile messages and mitigate the impact of complex biotic and abiotic factors from their surroundings  and pathways in the receiver plants to prepare a response that will match the upcoming stress  cultivars exposed to volatiles from another plant allocated more biomass to the roots  can distinguish volatiles from tomato (Lycopersicon esculentum) (host) and wheat (Triticum aestivum) (non‐host) plants and preferentially grow towards tomato plants because of specific compounds released such as β‐phellandrene, β‐myrcene and α‐pinene . Further studies conducted under field conditions are required to understand how plants discriminate between constitutive cues and periodical signals and how they use the available information during plant–plant interactions.Rapid adaption of plants to changes in the volatile emission of their neighbours demonstrates the important ecological role of touch‐induced VOCs as effective signals for both growth and defence synchronisation at the community level  induced response in neighbouring Arabidopsis thaliana, reducing root growth  promoted the growth and germination of their sympatric neighbours  exposed to onion (Allium cepa) contained four times higher concentrations of (E)‐nerolidol and ‐4,8,12‐trimethyl‐1,3,7,11‐tridecatetraene, which had a deterrent effect on green peach aphids (Myzus persicae)  were observed in barley when interacts with specific weeds. Of the 20 most prevalent weed species in cereals, only exposure to volatiles from lamb's quarters , black nightshades (Solanum nigrum) and thistles (Cirsium arvense and Cirsium vulgare) reduced aphid acceptance of exposed barley plants  or charlock (Sinapsis arvensis) induced the same effect in barley . In the same genotype mixtures tested in the field, the ladybird occurrence was significantly higher than that in the pure stands even before the aphid arrival .Detecting plants in the vicinity that are under herbivory attack can give a receiver plant an advantage  responded by the upregulation of defence genes as well as a change in volatile emissions  exposed to volatiles from damaged neighbours of the same chemotype triggered a much stronger antiherbivore defence than when exposed to volatiles from different chemotypes , male plants of the woody shrubs  released volatile signals that induced resistance in both male and female receivers, whereas VOC signals from the female plants only induced resistance in female receiver plants  plants emitted significantly higher amounts of constitutive and herbivore‐induced volatiles when growing in communities with the orchard grass (Dactylis glomerata) than in plants growing together with forbs (Geranium pratense)  had the capacity to modulate growth and defence strategies in response to the presence of conspecific or heterospecific Idaho fescue (Festuca idahoensis) neighbours. When grown with heterospecific neighbours, C. maculosa allocated more resources towards growth, but when grown with conspecific neighbours, C. maculosa invested more resources to defence  could deceive host plants to induce salicylic acid (SA)‐related defences while suppressing JA‐related defences and, through the release of volatile signals, could modify the quality of neighbouring plants for their offspring  to significantly increase the amount of total terpenoids in response to herbivory, revealing a strong synergy between higher temperatures and herbivory  covaried with the mean monthly temperature and the expected attack of a major root herbivore (Melolontha melolontha)  highlighted this possible mechanism during plant–plant interactions. Here, the emission rate of terpenoids was linked to trichome density in birch with an understorey of shrub (Rhododendron tomentosum)  have the ability to increase membrane depolarisation  provide a quick response to plant damage. When released from damaged plant cells, DAMPs activates the innate immune response of plants. In contrast to mammalian DAMPs, few binding receptors and pathways have been identified in plants. Herbivore‐induced plant volatiles can be classified as one type of DAMP that can be perceived by neighbouring plants , NtODC (a JA‐related gene) and NtACIII (an SA‐related gene) and tested 16 VOCs for their induction or inhibition properties. They found that each gene had a specific VOC with inducing capabilities. For a more specific understanding of induction, NtOsmotin was chosen for further analyses. β‐caryophyllene derivative‐linked beads were used to identify TPLs proteins, and two mechanisms of action have been proposed. The first is that β‐caryophyllene binds TPL, releasing an unknown co‐factor, which itself inhibits transcription factors when bound to TPL. The secondary mechanism by which β‐caryophyllene could induce expression of NtOsmotin is by binding TPL competitively, preventing it from binding to the unknown co‐factor   altered its growth pattern and emission of β‐cyclocitral to a degree six times higher than that of the negative control. This offered new insights into the evolution of plant–plant interactions via VOCs. Based on this study, it can be determined that VOCs are generally perceived as an essential tool in plant–plant communication and have been so for millions of years.From an evolutionary perspective, one of the oldest plant species is the bryophytes. The first evidence of plant–plant interactions via VOCs in non‐vascular plants was observed in a study showing that bryophytes can recognise their neighbours (Vicherová, Glinwood, Hájek, Šmilauer, & Ninkovic, A. thaliana has been shown (Xu et al., A long‐term strategy for the development of a plant's life is to provide the next generation with an advantage in terms of their specific surroundings. These exact evolutionary mechanisms are still under debate as this system is prone to false signalling, giving another surrounding plant a disadvantage. Attempts have been made to unravel VOC related signalling pathways in order to understand the complex networks of VOC emissions (Vivaldo et al., 5In nature, plants are simultaneously or in sequence exposed to a broad array of individual VOC cues and signals or a specific combination of these. Owing to the complex ecological effects they induce, plants must respond only to crucial ones to avoid compromising their growth and defence. This makes regulatory mechanisms extremely challenging, taking into consideration the trade‐off between cost and benefits of investment into adaptation. In addition, one or more important signals can be cancelled by the other one, leading plants to make wrong and costly decisions. In this respect, long‐distance signals are much easier to hinder, cancel or mask, so the real messages they carry are hidden or diluted to the level of non‐important cues. It is unclear whether and how plants can cope with a diverse array of VOCs in their environment, and which specific cues/signals may have higher priority to respond to. Deciphering the language of within‐ and between‐species plant communication requires studies under realistic ecological settings in which plant adaption responses determine plant fitness and survival at the individual and community levels.Recent studies on plant–plant communication by VOCs have revealed some metabolic pathways that plants activate to cope with biotic and abiotic stress. The molecular mechanisms of plant VOC perception remain largely unknown currently. Some topics have been addressed in recent years, showing that during plant–plant interactions, the regulatory mechanisms of perception are vastly complicated and highly specific, depending on the plant species, habitat and local plant community. It is clear that, from an evolutionary standpoint, VOC perception is a crucial tool for a plant to adjust its growth strategy and improve its fitness. Understanding the complex regulation of genetic and metabolic pathways could provide an important tool for commercial and sustainable agriculture. For this, further research must be conducted to understand the many factors that influence plant–plant interactions. Taking advantage of the natural mechanisms of how plants cope with biotic and abiotic stress would give us an edge in the rising demand for food production, especially since the genetic capacity for yield increase has been nearing its full potential. Studies so far have looked at the complex genetic regulation of VOC perception, mechanical structures and the area of proteomics to understand the metabolic pathways. Many open questions remain to be answered, and a general universal understanding of how different plants perceive and react to VOCs during plant–plant interactions remains to be found.The authors declare no conflict of interest.VN conceived the idea and revised the manuscript. DM, MR and VN contributed equally to manuscript writing and drafting. DM and MR prepared the figures.


Mycoplasma pneumoniae pneumonia (MPP) was made. The patient was successfully treated with a course of azithromycin and amoxicillin‐clavulanic acid. We suggest an awareness of diffuse pleuritis and a disseminated miliary nodular pattern as atypical manifestations of MPP, potentially attributable to immune modulation in the context of immunotherapy.A growing body of evidence suggests that check‐point inhibitors not only increase the overall risk of infections, but, due to an altered immune response, may also result in atypical manifestations. We report a case of a 38‐year‐old man with pleuritic chest pain, dyspnoea, fevers and a dry cough receiving combination ipilimumab and nivolumab immunotherapy for metastatic melanoma. Radiological findings demonstrated a diffuse increased fluorodeoxyglucose avidity of the thoracic pleura in addition to a disseminated miliary pattern of pulmonary nodularities. A subsequent bronchoscopy was macroscopically normal with unremarkable washings. In the context of a significantly elevated Mycoplasma serology, a diagnosis of  A novel case of disseminated miliary Mycoplasma pneumonia with invasive diffuse pleural involvement is reported. This is an atypical manifestation of a relatively common presentation likely due to altered immunity from concurrent check‐point inhibitor immunotherapy for metastatic melanoma. Mycoplasma pneumoniae (MP) is a common cause for respiratory infections, it is an uncommon cause of severe pneumonia in immunocompetent adults. Only 0.5%–2% of all MP pneumonia (MPP) presents with fulminant disease.Whilst A 38‐year‐old male was admitted to hospital with a 3‐day history of fevers, sore throat, dry cough and occipital headaches. He had recently received his second cycle of ipilimumab and nivolumab for metastatic BRAF wild‐type melanoma, with brain and lung metastases. The initial differential was that of an upper respiratory tract infection but was also treated with stat oral steroids to cover for immunotherapy pneumonitis. A subsequent computed tomography (CT) brain, chest x‐ray, blood examination and nasopharyngeal respiratory polymerase chain reaction (PCR) were all unremarkable. He was treated with intravenous fluids, ceftriaxone and doxycycline. The following day he was discharged home by the medical oncology team with a diagnosis of immunotherapy‐related fevers. He represented 1 week later with new‐onset bilateral pleuritic chest pain and lethargy, as well as worsening of his original symptoms. He was now mildly hypoxic with an oxygen saturation of 92% and had fine crackles on auscultation in bilateral lower lobes. A CT pulmonary angiogram revealed extensive widespread centrilobar and perilymphatic nodularities with consolidative and ground‐glass opacities around known bilateral lower lobe metastases  and immune check‐point inhibitor (ICI)‐induced pneumonia are two alternative diagnoses that have been shown to present with diffuse micro‐nodularity in the context of immunotherapy. Both these presentations differ from this reported case in that neither OP nor ICI‐induced pneumonia would have had any significant clinical response to antibiotic therapy alone, instead requiring prolonged courses of steroids to achieve clinical remission. Also, a significantly elevated initial Mycoplasma titre of >1:320 is considered diagnostic of an acute Mycoplasma infection, and the expect fall in the paired serology is also reflective of recent acute infection with an expected peak between 10 and 30 days.Extra‐pulmonary manifestations do occur but are rare and are believed to be immune mediated, rather than due to direct bacterial invasion.A growing body of evidence suggests that check‐point inhibitors, regardless of steroid use, increases the risk of infection through its immune modulation effect and altered host's response.In summary, we report a case of disseminated miliary pneumonia and clinically significant pleuritis with negative bronchial washings and markedly elevated Mycoplasma serology. This is in the context of concurrent check‐point inhibitor therapy for metastatic melanoma. We suggest that this is an atypical presentation of MP infection as a consequence of altered immunity.None declared.Appropriate written informed consent was obtained for publication of this case report and accompanying images.Both Daniel Bird and Maninder Singh had substantial contributions to the conception, design, analysis and interpretation of data for the case report. Daniel Bird was responsible for drafting the work, whilst Maninder Singh revised it critically for important intellectual content. Both gave final approval for the version to be published. Both agree to be accountable for all aspects of the work in ensuring that questions related to the accuracy or integrity of any part of the work are appropriately investigated and resolved.


Each miRNA is post‐transcriptionally processed into a duplex comprising two strands. One of the two miRNA strands is selectively loaded into an Argonaute protein to form the miRNA‐Induced Silencing Complex (miRISC) in a process referred to as miRNA strand selection. The other strand is ejected from the complex and is subject to degradation. The target gene specificity of miRISC is determined by sequence complementarity between the Argonaute‐loaded miRNA strand and target mRNA. Each strand of the miRNA duplex has the capacity to be loaded into miRISC and possesses a unique seed sequence. Therefore, miRNA strand selection plays a defining role in dictating the specificity of miRISC toward its targets and provides a mechanism to alter gene expression in a switch‐like fashion. Aberrant strand selection can lead to altered gene regulation by miRISC and is observed in several human diseases including cancer. Previous and emerging data shape the rules governing miRNA strand selection and shed light on how these rules can be circumvented in various physiological and pathological contexts.RNA Processing > Processing of Small RNAsRegulatory RNAs/RNAi/Riboswitches > Regulatory RNAsRegulatory RNAs/RNAi/Riboswitches > Biogenesis of Effector Small RNAsThis article is categorized under: Summary of miRNA strand selection and regulatory mechanisms. Three major points of regulation for miRNA strand selection (1) altered miRNA processing by the Microprocessor complex (not shown) or Dicer, (2) remodeling of the miRNA duplex by nontemplated RNA modifications such as uridylation or A‐to‐I editing, and (3) changes in the strand preference of Argonaute. Once presented with a miRNA duplex, Argonaute makes a binary choice to load one miRNA strand and discard the other strand from miRISC. Alternate Argonaute programming leads to shifts in the target profile of miRISC based on the seed sequence of the loaded strand and is sometimes associated with human diseases. ArgonauEach strand of the miRNA duplex is destined for different fates. One of the duplexed miRNA strands is selected and anchored into the Argonaute protein, while the other strand is unwound from the complex and is subject to degradation Figure . The 5′ The Argonaute‐loaded miRNA programs the gene target specificity of the miRISC, as nucleotides 2‐7, referred to as the miRNA “seed” sequence, are mainly responsible for determining the target mRNA identity through base pairing  of each strand and the relative thermodynamic stability (TS) of the two ends of the duplex Figure . StructuC. elegans, single amino acid substitutions within the MID (G553R) and PIWI  domains of the miRNA Argonaute ALG‐1 (Argonaute‐like Gene) reverse the strand selection of certain miRNAs  enzymes  enzymes exhibit reversed strand selection for some of the miRNAs . A‐to‐I editing has been shown to influence miRNA strand selection in mice and human cells  was proposed to sense the relative TS of perfectly‐paired siRNA duplexes with R2D2 binding to the more stable end thereby passing the less stable end to Ago2, suggesting a direct role of Dcr‐2/R2D2 in strand selection  progression, alters the sequence of miR‐146a* (3p) strand, and likely leads to changes in target repression, contributing to PTC  expression is down‐regulated in nonsmall cell lung cancer (NSCLC)  and miR‐21* (3p) are up‐regulated and each is required for cell survival  altered processing of the miRNAs by RNAse III endonucleases generating templated isomiRs, (b) nontemplated miRNA isomiRs generated through deamination (A‐I editing) or uridylation, which remodel the miRNA duplex, and (c) altered strand preference of Argonaute Figure . NotableIn vitro experiments have provided and will continue to provide important advances in understanding small RNA loading. However, the rules governing miRNA strand selection were determined using a limited number of purified components; whether these rules are sufficient to drive strand selection in vivo has not been extensively explored. At first glance, a significant number of miRNAs do not follow either the 5′ nucleotide or TS rules (see Figure The authors have declared no conflicts of interest for this article.Jeffrey Medley: Formal analysis; visualization; writing‐original draft; writing‐review and editing. Ganesh Panzade: Data curation; writing‐review and editing. Anna Zinovyeva: Conceptualization; funding acquisition; resources; supervision; visualization; writing‐original draft; writing‐review and editing.Anatomy of RISC: how do small RNAs and chaperones activate Argonaute proteins?The organization and regulation of mRNA‐protein complexes


Dear Editoret al. in regards to our recently published paper: Pokhrel et al. Clinical validation of ring‐mounted halcyon Linac for lung SBRT: comparison to SBRT‐dedicated C‐arm Linac treatments. J Appl Clin Med Phys 2021; 22(1); 261‐270.We read with interest the letter to the editor drafted by Sarkar et al. do not specifically pertain to our paperThe author’s study immediately significantly deviates from our paper as they used a 6 MV flattened beam (much higher mean energy and corresponding PDD) with significantly different beam characteristics. Alternatively, we used the closest possible beam profile and characteristics via the 6MV‐FFF beam on C‐arm TrueBeam Linac for the validation of single‐energy 6MV‐FFF Halcyon for lung SBRT treatments. This would greatly affect the feasibility of meeting RTOG‐0618 requirements.In terms of the dosing prescription scheme, the author states their planning study was for 60 Gy in five fractions using RTOG‐0618 criteria. We believe that the most common prescription of the RTOG‐0618 trial is a 54 Gy in three fractions scheme.In our study and at our institution, both lung SBRT plans were normalized to ensure 95% of the PTV is covered by 100% of the prescribed dose with a 120–130% hot spot in the GTV. This is accomplished by prescribing the dose to the tumor periphery with a sharp penumbra using the 70–80% isodose line. This prescription effectively constraints the hotspot to the center of the GTV and provides a sharp dose gradient for heterogenous SBRT dose distribution. Ultimately, this reduces the dose to the normal lung as intended in the RTOG protocols. In contrast, the author presents an example of a more homogenous dose distribution based on their reported normalization conditions. By creating a more homogenous dose distribution, normal lung dose will increase because of the inherent increase in intermediate dose spill that our institution avoids. To compound this normalization condition, the tumor size was not reported by the author for their two specific cases where a larger tumor size could explain the violation of RTOG‐0618 criteria observed.To characterize the Novalis non‐coplanar lung SBRT capabilities with respect to the Halcyon, more information is required in the letter to the editor to not confuse any potential readers. The author mentioned that 40% NTX plans were non‐coplanar beams; however, they did not mention the technique deployed  nor the couch angles chosen for these static/arc geometry beams. We do acknowledge that due to collision issues there are limited angles available for non‐coplanar geometry on a C‐arm Linac for lung SBRT. However, its unwise to infer the angles used for their respective NTX plans despite the comment of a “similar” set‐up (line 11 of the author’s letter) to our study. It is understood that using an exotic non‐coplanar beams/arcs arrangement will yield high‐quality and RTOG compliant treatment plans. Unfortunately, this does not guarantee for the plan deliverability as it may render collision issues  or impractical longer treatment delivery times for select distressed pulmonary cancer patients. It is for these reasons more information about the non‐coplanar NTX plans must be included in the author’s letter as it cannot be certain uncommon beam geometry was deployed for these compliant plans. This is one of the reasons the author’s claim that the Halcyon Linac is uncapable of meeting RTOG‐0618 criteria for these 2 patients is clouded.It is acknowledged by the author that the NTX flattened beam significantly differs from the Halcyon beam characteristics. This includes differences in mean energy, PDD, beam profile, and multi‐leaf collimators (MLC) characteristics. The Halcyon Linac is equipped with a next‐generation SX2 MLC with improved and differing values for MLC size , less leakage, and transmission dose of <0.5%. Therefore, we used the closest possible 6MV‐FFF beam energy of TrueBeam Linac (with standard 120 Millennium MLC) to compare our Halcyon validation plans for lung SBRT. We believe that this will better characterize the mechanical improvements or limitations of the Halcyon V2.0 for lung SBRT treatments. Without a detailed dosimetric study, we believe it is not appropriate to make any comparison with our paper as using an FFF‐beam could potentially increase the dose coverage at the tumor periphery and reduce the out‐of‐target normal tissue dose.Halcyon gantry rotation speed is “4 RPM” not “2 RPM.”We acknowledge and agree with the author that maximal dose rate differences between modalities may play a role in final beam‐shaping capabilities in the VMAT planning of these lung SBRT treatments. In our Halcyon lung SBRT validation study with the RTOG‐0813 protocol, we use the TrueBeam 6MV‐FFF beam with a maximal possible dose rate setting of 1400MU/min vs 6MV‐FFF on Halcyon with 800MU/min. Unfortunately, we cannot be certain the author deployed an inverse planning technique for their NTX plans as no specific mention of their planning technique was included in the letter to the editor.In their example case, for the same prescription, the author reported almost 80% higher total MU with NTX compared to the Halcyon plan. We find this increase extremely concerning if the same technique is used between the modalities. This may indicate a fundamental issue with the study described in the letter to the editor. For the given prescription and similar planning technique, we believe that non‐coplanar geometry should give similar or even less total MU, in general, due to relatively less chance of beams or arcs overlapped in the target. Our experience for lung SBRT is that for the same prescription and identical target coverage, we get similar MU, similar intermediate dose‐spillage, but slightly higher low dose volume with 6MV‐FFF Halcyon compared to 6MV‐FFF TrueBeam. We reported this in our paper via slightly higher volumetric doses to the OAR such as dose to 10 cc of skin.We agree with the author that, unlike intracranial SRS, there is a limited scope of non‐coplanar geometry in the extracranial region due to the gantry collision issues. Clinics that use a advanced treatment planning system with VMAT or IMRT optimization may deploy additional approaches such as optimal values of normal tissue objectives, OAR avoidance feature or sectors avoidance (in VMAT) to further compensate for lack of full angular capabilities in these treatments and further spare the OAR. Author needs to clarify the details of their treatment planning approach.In our paper, in addition to achieving the clinically acceptable and RTOG‐0813 compliant plan quality for select lung SBRT patients, we present a discussion about Halcyon delivery efficiency. Our concern with the “4π algorithm”We strongly disagree with the author’s unproven claim that the Halcyon planning technique is limited. Based on our clinical experience and the currently available literature,As the author suggested, a small yaw rotation to the couch and a small gantry tilt is similar to some advanced CT simulator, however, to incorporate this feature on Halcyon, we think that would need a major structural re‐design of the current Halcyon Linac by the Varian Medical Systems . Therefore, the author’s conclusion suggesting: “it will result in a better dosimetric outcome” is premature and yet to be demonstrated (or designed).We appreciate the interest in our recently published paper by the author, however, after carefully reviewing this draft, we found several flaws in the letter to the editor. We strongly feel the assertions presented by Sakar Therefore, we believe that the generation of a high‐quality clinical lung SBRT plan does not only depend on the coplanarity (or lack‐thereof) beam geometry on a case‐by‐case basis, but also depends upon the fine details of every aspect of the treatment planning approach and the patient selection that we described above. To conclude, based on the deficiencies as noted above, the author’s letter to the editor is not relevant or consistent with our published paper on the Halcyon Linac’s lung SBRT capabilities. This includes fundamental differences between our detailed study and the author’s very limited study in terms of concept, methodology, and results that we believe provide clinical value to the fellow Halcyon users. Additionally, due to the incomplete and inconsistent description of the author’s limited study, it may create confusion and ambiguity concerning the Halcyon’s ability  to commission and implement the SBRT protocols in the clinic for the prospective patient’s treatment.


However, despite the tremendous progress that has been made in this field, many misconceptions remain among much of the broader scientific community about the manner in which miRNAs function. In this review, we focus on miR‐33, one of the most extensively studied miRNAs, as an example, to highlight many of the advances that have been made in the miRNA field and the hurdles that must be cleared to promote the development of miRNA‐based therapies. We discuss how the generation of novel animal models and newly developed experimental techniques helped to elucidate the specialized roles of miR‐33 within different tissues and begin to define the specific mechanisms by which miR‐33 contributes to cardiometabolic diseases including obesity and atherosclerosis. This review will summarize what is known about miR‐33 and highlight common obstacles in the miRNA field and then describe recent advances and approaches that have allowed researchers to provide a more complete picture of the specific functions of this miRNA. miRNAs have emerged as critical regulators of biologic processes and important therapeutic targets for numerous diseases. This review focusses on miR‐33 as an example, and describes the obstacles, recent advances and new approaches in the miRNA field. AtherosclerosisVascular disease characterized by the progressive accumulation of lipids and inflammatory cells in the subendothelial space of large arteries.Binding site mutant (BSM)Targeted modifications of miRNA binding sites to selectively disrupt specific miRNA/target interactions.Cholesterol EffluxProcess by which peripheral tissues transfer intracellular cholesterol onto extracellular acceptors like high‐density lipoprotein cholesterol (HDL).High‐density lipoproteins (HDL)Lipoprotein particles that promote the cellular cholesterol efflux from cells and deliver cholesterol to the liver and steroidogenic tissues via selective lipid uptake.microRNA (miRNA)Small non‐coding RNA that negatively regulates protein expression by promoting mRNA degradation or impairing protein translation.pH low insertion peptides (pHLIP)Constructs designed to deliver small amounts of nuclear material only to acidic microenvironments.Reverse Cholesterol Transport (RCT)Process by which cholesterol is removed from peripheral tissues and transported to the liver for removal or redistribution.RNA‐induced Silencing Complex (RISC)Multiprotein complex that facilitates targeted inhibitory activity of small regulatory RNAs like miRNAs and siRNAs.et al,et al,et al,et al,et al,et al,Since the structure of DNA was uncovered in 1953, the central dogma of biology—that cellular functions are carried out by transcription of DNA into RNA, which is then translated into the proteins that carry out physiologic functions—has remained largely unchanged. Under this theory, regions of the DNA that do not code for proteins are largely non‐functional. However, with the discovery of miRNAs in 1993  that can be incorporated into the RNA‐induced silencing complex (RISC). Once incorporated into the RISC, miRNAs facilitate binding of a distinct subset of mRNA targets by targeting specific sequences, generally within the 3’ untranslated regions (3’UTR), leading to either mRNA degradation or impaired translation Ambros, . One of The combination of these factors has made determining the specific mechanisms by which miRNAs exert their effects very challenging. For many years, researchers have relied on a limited toolset with which to identify and validate miRNA targets. A number of different target prediction algorithms have been developed that can be used identify which mRNAs are likely targets for a specific miRNA, and this targeting can then be validated using 3’UTR luciferase assays. However, assessing the likelihood that a miRNA target is involved in mediating a specific function has largely relied on associations between mRNA/protein levels and alterations in miRNA expression. In recent years, a great deal of progress has been made in the development and implementation of new tools and techniques that allow researchers to directly assess the binding of mRNAs to the RISC and specific miRNAs, determine the relative impact of individual miRNA/target interactions, and evaluate the specific role of miRNAs in different tissues and/or cell types. These new strategies will help address many of the main obstacles facing the development of miRNA‐based therapies by improving our ability to determine the specific mechanisms by which miRNAs exert their effects and assess potential detrimental outcomes of miRNA‐based therapies.miR‐33 is one of the most well‐studied miRNAs and is an excellent example of the potential of miRNA‐based therapeutics to treat a variety of different human diseases, the possible hazards that may be involved in miRNA‐based therapeutic approaches, and the techniques that are being developed to try to better understand and minimize these risks. In this review, we will summarize the work that was done to characterize the role of miR‐33 in metabolism, atherosclerosis, and other conditions. We will then describe in detail the recent advances that have been made in understanding the specific functions of miR‐33 with a focus upon the utilization of new tools for the study of miRNA biology, including tissue‐specific mouse models, new approaches to disrupt specific miRNA/target interactions, and targeted miRNA therapies.Proper control of lipid homeostasis is crucial for the maintenance of human physiology and health , whereas insulin‐responsive SREBP1c is more selective for genes involved in fatty acid metabolism and lipogenesis  , cholesterol conversion to bile acids (CYP7A1), and cholesterol secretion into bile (ABCG5/G8)  are nuclear hormone receptors that form active heterodimers with retinoid X receptors (RXRs) and are activated by a variety of sterols, including oxysterol intermediates that form during cholesterol biosynthesis  that are encoded within introns 16 and 17 of the human SREBF2 and SREBF1 genes, respectively   risk in human epidemiological studies has led to a great deal of work exploring the potential of inhibiting miR‐33 in prophylactic and therapeutic murine models of atherosclerosis. Inhibition or deletion of miR‐33 in Western diet (WD)‐fed et al,in vitro and in vivo. Initially, it was shown that miR‐33 can reduce cholesterol efflux to ApoA1 and HDL by modulating the expression of ABCA1 and ABCG1 (rodents only) in macrophages. Importantly, inhibition of endogenous miR‐33 results in increased expression of the ABCA1 protein and a concomitant increase in cholesterol efflux, indicating a physiologic role for this miRNA in regulating ABCA1 expression , the process by which mammalian cells orchestrate the removal of excess cholesterol from peripheral tissues to the liver , as well as their transcriptional regulators (Foxo3 and Tfeb) , vascular smooth muscle cells (VSMCs), and/or macrophages. As discussed, miR‐33 is an important regulator of fatty acid metabolism and cellular bioenergetics and can also play an important role in regulation of insulin signaling and glucose metabolism  studies have raised questions about the possible deleterious metabolic consequences of antagonizing miR‐33 , Horie et al showed that both BM‐specific and whole‐body loss of miR‐33 resulted in reduced lipid accumulation in atherosclerotic plaques, but the effects on plaque size were more pronounced in animals lacking miR‐33 globally. These findings suggested that both liver and macrophages play an important role in mediating the effects of miR‐33 on atherosclerosis  transplants from miR‐33‐deficient animals into ApoE‐deficient mice , providing further evidence that loss of miR‐33 in the liver can protect against the development of hepatic fibrosis ‐ and macrophage (LysM‐Cre)‐specific miR‐33 knockout models. Adipocyte‐specific deletion of miR‐33 did not result in any differences in body weight, regulation of glucose homeostasis, or response to a lipid challenge in HFD‐fed animals  mouse model, we were able to demonstrate that selective disruption of the ability of miR‐33 to target ABCA1 is sufficient to enhance macrophage reverse cholesterol transport and reduce atherosclerotic plaque burden in vivo .https://diabetes.org: An international professional society that supports research on processes fundamental to diabetes (American Diabetes Association).https://omim.org/entry/612156: A resource page on miR‐33 from the Online Mendelian Inheritance in Man online catalog of human genes and genetic disorders.http://www.mirbase.org: A searchable database of published miRNA sequences and annotation.http://www.targetscan.org: A searchable database of predicted miRNA target genes.