A 56-year-old man with a previous right nephrectomy for multiple papillary renal cell carcinomas (pRCC) underwent a follow-up CT scan. Using a dual-layer dual-energy CT (dlDECT), we demonstrated the presence of a small amount of fat in a 2.5 cm pRCC that mimicked the diagnosis of angiomyolipoma (AML). Histological examination demonstrated the absence of macroscopic intratumoral adipose tissue, showing a fair amount of enlarged foam macrophages loaded with intracytoplasmic lipids. The presence of fat density in an RCC is an extremely rare occurrence in the literature. To our knowledge, this is the first description using dlDECT of a minimal amount of fat tissue in a small RCC due to the presence of tumor-associated foam macrophages. Radiologists should be aware of this possibility when characterizing a renal mass with DECT. The option of RCCs must be considered, especially in the case of masses with an aggressive character or a positive history of RCC. The detection of fat-density components represents a key diagnostic criterion for the computed tomography (CT) characterization of a renal mass A,B. The We describe the case of a 56-year-old man with a history of right nephrectomy due to papillary renal cell carcinoma who underwent a follow-up CT scan. Using a dual-layer DECT (dlDECT) , we confirmed the presence of a small amount of fat in a 2.5 cm papillary renal cell carcinoma (pRCC) that mimicked the diagnosis of AML. Histological examination revealed a significant number of foam macrophages and the absence of macroscopic intratumoral adipose tissue .Histological examination demonstrated the absence of macroscopic intratumoral adipose tissue, showing a fair amount of foam macrophages. As with other solid tumors, RCCs comprise a heterogeneous microenvironment of both malignant and normal stromal cells that contain a large number of macrophages, originating from blood monocytes attracted by chemokines and cytokines produced by tumor cells. Tumor-associated macrophages were demonstrated to directly stimulate tumor cell proliferation and angiogenesis . They ar


The positional relationship between the coronary artery and the transcatheter heart valve (THV) commissure is crucial for coronary access following transcatheter aortic valve implantation (TAVI). The ALIGN TAVR study demonstrated optimizing valve alignment using a marker on the delivery system or THV commissural post . HoweverThe en-face view is useful for comprehending the short-axis orientation in fluoroscopy . NavitorThis can help recognize short-axis orientation, although it might be challenging to create a true en-face view because the Navitor THV is still connected to the delivery system at 80% deployment. Since Navitor has a recapture function, if misalignment is discovered at 80% deployment in the en-face view, the need for recapture and rotation of the delivery catheter to accomplish commissural alignment can be discussed. In addition, the next-generation Evolut FX system may be also able to do the same assessment because it has fluoroscopic markers that indicate the commissures at the left ventricle edge.Supplementary file1 Aortography with the en-face view in the aligned case. (MP4 312 KB)Supplementary file2 Aortography with the en-face view in the misaligned case. (AVI 2651 KB)Below is the link to the electronic supplementary material.


Incorrect FundingIn the published article, there was an error in the Funding statement. The Funding statement is not full. Original incorrect text: “This work was supported by the Deutsche Forschungsgemeinschaft  cluster of excellence EXC 2177/1. Part of these results is part of the project that received funding from the European Research Council (ERC) under the European Union's Horizon-ERC Program (Grant agreement READIHEAR No. 101044753-PI: WN).” The correct Funding statement appears below.


Tuberculosis remains a leading infectious cause of death in resource-limited settings. Effective treatment is the cornerstone of tuberculosis control, reducing mortality, recurrence and transmission. Supporting treatment adherence through facility-based observations of medication taking can be costly to providers and patients. Digital adherence technologies (DATs) may facilitate treatment monitoring and differentiated care. The ASCENT-Ethiopia study is a three-arm cluster randomised trial assessing two DATs with differentiated care for supporting tuberculosis treatment adherence in Ethiopia. This study is part of the ASCENT consortium, assessing DATs in South Africa, the Philippines, Ukraine, Tanzania and Ethiopia. The aim of this study is to determine the costs, cost-effectiveness and equity impact of implementing DATs in Ethiopia.n = 150). We will conduct a societal cost-effectiveness analysis using Bayesian hierarchical models that account for the individual-level correlation between costs and outcomes as well as intra-cluster correlation. An equity impact analysis will be conducted to summarise equity efficiency trade-offs.A total of 78 health facilities have been randomised (1:1:1) into one of two intervention arms or a standard-of-care arm. Approximately 50 participants from each health facility will be enrolled on the trial. Participants in facilities randomised to the intervention arms are offered a DAT linked to the ASCENT adherence platform for daily adherence monitoring and differentiated response for those who have missed doses. Participants at standard-of-care facilities receive routine care. Treatment outcomes and resource utilisation will be measured for each participant. The primary effectiveness outcome is a composite index of unfavourable end-of-treatment outcomes  or treatment recurrence within 6 months of end-of-treatment. For the cost-effectiveness analysis, end-of-treatment outcomes will be used to estimate disability-adjusted life years (DALYs) averted. Provider and patient cost data will be collected from a subsample of 5 health facilities per study arm, 10 participants per facility  PACTR202008776694999. Registered on 11 August 2020, The online version contains supplementary material available at 10.1186/s13063-023-07289-x. Tuberculosis remains a leading infectious cause of death, disproportionately affecting people living in low-resource settings . IneffecHistorically, tuberculosis programmes emphasised directly observed treatment short-course strategy (DOTS) in high-burden countries . The corIn Uganda, a step-wedged trial of medication sleeves, where patients call a toll-free number revealed when a medication dose is removed from the blister pack (99DOTS), found no evidence of improved treatment success during the intervention period . This stThe objective of this study is to estimate the cost-effectiveness of the interventions compared to the standard of care. The secondary objective of the study is to estimate the distribution of the costs and outcomes of the intervention by household socio-economic position. The objectives of the trial are to evaluate whether the implementation of (i) digital pillboxes or (ii) medication labels with daily monitoring using a web-based platform, followed by a tailored response by healthcare workers to adherence data, decreases the proportion of adult pulmonary drug-susceptible tuberculosis patients with unfavourable outcomes compared to the standard-of-care. The primary trial outcome will be assessed as a composite outcome of poor end-of-treatment outcomes  or recurrence of tuberculosis disease within six months of the end of treatment. Secondary objectives include to (a) describe longitudinal technology engagement, (b) describe the fidelity to the adherence tools, (c) project the epidemiological impact of intervention scale-up and (d) estimate the cost-effectiveness of the intervention compared to the standard-of-care.We are conducting a three-arm pragmatic cluster randomised trial where the unit of randomisation is health facilities . A totalWe plan to enrol 3900 patients (≥ 18 years old) with pulmonary DS-TB in the trial (50 per facility) over a 12-month period and follow them up 12 months after treatment initiation. Participants with bacteriologically confirmed tuberculosis at enrolment, a successful end of treatment outcome and able to produce sputum, will provide a sputum sample for culture approximately 6 months after the end of treatment to measure disease recurrence among trial participants. Primary study outcome is a composite unfavourable outcome of (i) treatment lost-to-follow-up, death, and treatment failure and (ii) disease recurrence within 6 months of end of treatment.The intervention is DATs, the digital pillbox called EvriMed or medication labels linked to a data platform and differentiated care based on adherence data. Each dose taken by patients is logged on the platform either automatically when the digital pillbox is opened, or when the patient texts a code on the dose label to a dedicated toll-free number. Healthcare workers have access to the ASCENT data platform on a web-based platform or mobile phone app, which allows treatment adherence to be evaluated and support updated. A flow diagram indicating decision points and actions is shown in Fig. In the standard-of-care arm, doses are retrospectively recorded in the facility tuberculosis record during treatment visits. Prior to the COVID-19 pandemic, patients in the intensive phase of treatment received DOT. Currently, patients are self-administering medication for 1 to 2 weeks unless the providers’ assessment identifies adherence as a concern. If clinic appointments are not attended, patients will be phoned to provide adherence counselling. If further clinic appointments are missed, a community health worker will conduct a home visit.In the intervention arms of the study, one missed clinic appointment will be followed up telephonically. Following two consecutive missed clinic visits, a community health worker will visit the home. After the first 2 weeks, for each missed dose, patients receive a same-day automated reminder SMS. If two or more consecutive doses are missed, the patient will receive a telephone call. More than five missed doses result in a home visit and if doses are persistently missed (> 14 consecutive doses or multiple times missing 7 doses), discontinuation from the DAT intervention and switch to daily DOT under healthcare workers’ observation will be considered.Possible harms due to study participation include the risk of inadvertent disclosure of patients’ tuberculosis status because of the use of digital pillboxes or receiving SMS notifications. These harms are monitored by keeping a health facility social harm register. Inadvertent disclosures are recorded at the 12-month follow-up and reviewed by the ASCENT Trial Advisory Group .Adults (≥ 18 years old) receiving pulmonary DS-TB treatment at the trial facilities are screened for eligibility by facility healthcare workers, and informed consent is requested from eligible patients. Participant exclusion criteria include patients who are expected to migrate out of the health facilities’ catchment area in the next 12 months and patients receiving in-patient or palliative care. Participants who withdraw from the study or stop using a DAT will continue to receive routine care and asked if their adherence and treatment outcome data may be collected.The primary analysis will be an intention-to-treat (ITT) analysis, where the costs and effects of all participants enrolled in the trial are included, excluding participants whose diagnosis are changed to ‘no TB’, are started on MDR treatment, or are transferred out ≤ 28 days of treatment start. As a sensitivity analysis, we will conduct a per-protocol (PP) analysis that excludes participants in the intervention arm who opted out of using DAT during the treatment period, started DAT ≥ 28 days after treatment initiation, was withdrawn from using DAT due to poor network, lost phone or poor adherence.Sample size estimation for the primary trial outcomes considered the clustered study design and pair-wise comparisons of the standard-of-care—to the intervention arms . ProgramData collection for the study is expected to be completed by June 2023, and the results of the trial analyses available in October 2023. There are no interim analyses planned.Treatment outcomes and events will be collected for 3900 trial participants. Estimates of provider and patient costs will be collected from a sub-sample of 15 health facilities, 5 facilities per study arm and 10 participants per facility, preserving the empirical correlation between effects and costs and therefore providing a more accurate estimate of the uncertainty interval of the average cost-effectiveness estimate , 18. Hean = 150), in the intensive or continuation phase of treatment. Provider unit costs will be collected from subsample facilities using two facility surveys (facility surveys A and B) completed by the facility TB focal person over a period of 6 weeks. Surveys will be used to record time spent on tuberculosis treatment-related tasks with ten consecutive participants and the amount of time spent weekly on adherence support (including home visits). A facility cost data collection tool will be used to collect provider cost data. The cost of implementing the intervention will be estimated using KNCV records on the cost of purchasing and distributing DATs. The cost of training and supporting healthcare workers in implementing DATs will be estimated using KNCV training logs and staff interactions with health facilities logged on the ASCENT platform.Baseline participant demographics and household socio-economic position (SEP) will be collected at study entry for all trial participants. Enrolled participants’ treatment outcomes  will be abstracted from the facility tuberculosis treatment register by health facility staff. Twelve months after treatment initiation, trial participants with a successful end-of-treatment outcome will be contacted to confirm their outcomes. Health facility visits logs completed by healthcare workers at the subsample health facilities will be used to record all visits made by trial participants. Direct- and indirect patient unit costs will be collected from ten participants per facility  costs and effectiveness endpoints will be correlated, 2) data will be hierarchical because of the randomisation of facility clusters in the trial whereby each cluster is distinct from the other clusters and data within clusters are broadly similar, (3) cost data tend to be right skewed and therefore model assumptions of normally distributed data will not hold, (4) structural zero costs where some data points are expected to be zero, and (5) partially observed data because costs are collected from a subsample of facilities , 20. We  data wil costs anBaseline participant characteristics that will be summarised and compared between study arms are shown in Table We will use an ingredient costing approach to calculate the provider cost of human resources, training, and implementing the intervention in 15 of the 78 trial facilities . OverheaM) complete datasets [Patient costs include the direct costs, the indirect costs, and the value of assets sold due to hardship experienced due to illness or accessing treatment. Patients’ time will be valued using self-reported monthly income and country-specific minimum wage in the sensitivity analysis. The total patient cost will be estimated as the sum of direct costs, the value of time lost due to accessing treatment and the value of any assets sold. We will report the number of patients experiencing catastrophic costs as the number of patients for whom total patient costs exceed 10% of their self-reported income . The cosdatasets .M) complete datasets, costs and effectiveness will be modelled jointly using Bayesian hierarchical models to account for the clustered nature of the data. Random effects will be specified to allow for individual-level variances that differ across clusters. The cost and effectiveness variables will be modelled using zero-inflated Poisson regression models, therefore not assuming that zero and non-zero values come from the same data-generating processes [M submodels [Statistical models will be used to estimate the differences in costs and outcomes between study arms and to analyse associations between patient-level factors and costs/outcomes. For each of the 10  by a unit cost  . The valThe mean differences in costs and DALYs with be estimated through pair-wise comparison of each study arm to the standard-of-care study arm, and cost per DALY averted ranked from the lowest to highest number. An intervention will be considered cost-effective if (i) less costly and more effective than the standard-of-care or (ii) more costly and more effective but the incremental cost-effectiveness ratio (ICER) is below the cost-effectiveness threshold (CET).CETs are country- and payer-specific, representing the opportunity costs of investing in alternative health interventions –36. We wAn asset index representing household SEP will be created using principal components analysis (PCA) of 23 variables measuring various dimensions of poverty. The questions are based on previous studies conducted in Ethiopia and include (i) education, (ii) frequency of household income, (iii) marital status, (iv) access to sanitation, (v) cooking fuel, (vi) household crowding and (vii) ownership of consumer goods, land and livestock –45. The Traditional cost-effectiveness analyses implicitly assume that costs and outcomes are distributed equally within the cohort irrespective of household socio-economic position (SEP). Here, we extend the decision framework to include measures of the distribution of the costs and intervention effectiveness by household SEP, by estimating the cost and illness concentration indices , 47. TheSubgroup analyses will be conducted to investigate how the intervention effect on outcomes and costs changes in subgroups defined by SEP, gender, time on treatment, health service utilisation and geographic region. Planned sensitivity analyses are summarised in Table The MCMC output from the Bayesian model will be used for post-estimation analysis of the parameter uncertainty in the decision analysis. We will estimate whether the intervention(s) would be cost-effective against the standard-of-care at a range of CETs. The results of this analysis will be presented on a cost-effectiveness acceptability frontier, showing the probability that a simulation will be cost-effective for each intervention against a range of CETs .The analysis described here is a within-trial cost-effectiveness—and equity impact analysis evaluating the distributional cost-effectiveness of two digital adherence technologies (DATs) for support of tuberculosis treatment adherence. Both DATs collect dose-by-dose adherence data which can then inform healthcare workers to intervene and support tuberculosis treatment adherence without the need to wait for clinic visits to address challenged and encourage adherence. To date, few studies have produced estimates of effectiveness, the patient and patient costs incurred for dose-by-dose adherence support interventions using randomised studies . FurtherThis trial was implemented at the time of the COVID pandemic, which resulted in changes in the ways that people access healthcare services in Ethiopia and globally. There is some evidence that routine visits to healthcare facilities have reduced due to the pandemic . What isThere are a range of approaches currently used to assess the distributional (equity) impact of national decision-making , 47, 58.Recruitment to the run-in phase of the trial started in December 2020, followed by the main phase in June 2021 and recruitment will continue until June 2022. Follow-up is expected to continue until June 2023. This analysis plan was first drafted at the end of the run-in phase and completed prior to data analysis.Additional file 1: S1 Text.


Purpose: This study aimed to investigate the lower limb inter-joint coordination and variability during Tai Chi movements compared with normal walking in older adults.Methods: A total of 30 female Tai Chi practitioners (70.9 ± 5.2 years) were recruited in this study. Herein, each participant performed three trials of the normal walking and Tai Chi movements. The lower limb kinematics data were collected with Vicon 3D motion capture system. The continuous relative phase (CRP) includes both spatial and temporal information of two adjacent joints, which was calculated to assess the inter-joint coordination of lower limbs. Coordination amplitude and coordination variability were assessed with mean absolute relative phase (MARP) and deviation phase (DP). MANOVOA was used to analyze inter-joint coordination parameters between different movements.Results: The CRP values of hip-knee and knee-ankle segments in the sagittal plane of the Tai Chi movements changed frequently. The MARP values of the hip-knee (p < 0.001) and knee-ankle segments (p = 0.032) as well as the DP values of the hip-knee segment (p < 0.001) were significantly lower in Tai Chi than in normal walking.Conclusion: More consistent and stable inter-joint coordination patterns of Tai Chi movements found in this study may be one of the critical factors that Tai Chi could be a suitable coordinated exercise for older adults. Falls are a major health problem for older adults worldwide. The incidence and number of falls in elderly women were about twice as high as those in males in the United States . Falls cThus, Tai Chi is a promising method. As a low-speed, low-impact exercise, Tai Chi is a traditional Chinese exercise that is popular among older adults. In addition, Tai Chi is beneficial for retaining or regaining proper balance and coordination for older people . A six-wContinuous relative phase (CRP) is a commonly used technique to investigate movement coordination. CRP is derived from the angular velocity–angle phase diagram of two joints or segments changing with time, and it is associated with information about afferent joint receptors . The magHowever, the multi-joint coordination of lower limbs is lacking during typical Tai Chi movements, thus more scientific research is needed to understand the basic mechanisms and principles of how Tai Chi works. Therefore, this investigation aimed to describe and quantify the lower limb inter-joint coordination and coordination variability during Tai Chi movements in older adults and explain the beneficial effect of Tai Chi on coordination and proprioception compared with normal walking. For comparison, the normal walking gait is chosen because it is the most common gait pattern in daily activities in elderly populations. In addition, the brush knee and twist step (BKTS) is a typical basic Yang-style Tai Chi movement and continuous forward gait, with gait phases similar to normal walking . It was A total of 30 healthy female Tai Chi practitioners  were recruited in this study. Inclusion criteria are as follows : (1) ageWith a sample size of 30, the power of all statistical comparisons in this study was no less than 0.80. It was reported that 15–20 participants were sufficient in estimating the reliability of a quantitative variable .A walkway was constructed for data collection . Two forThe participants’ anthropometric data were collected before the tests. Herein, an experienced research lab technician and participants were blinded to conduct data collection. Then, each participant was asked to wear a tight black uniform and shoes provided by the laboratory. A total of 41 infrared reflective markers with a diameter of 14 mm were attached to the skin or clothes of each participant with double-adhesive tapes. Afterward, before data collection, the participants were instructed to perform 5 min warm-up to familiarize themselves with the experimental set.The foot movements during the BKTS are described as follows: forward movement, the right foot makes contact with the ground first, the left foot then steps forward, and the right foot leaves the ground at the end . Then, tAs determined through a kicking ball test, the right leg was the dominant limb among all participants. In addition, the left and right cycles have quasi-identity during most Tai Chi movements . Therefox-axis) and velocities (ω) (y-axis). Angular positions and velocities were normalized using the following equations:i and ωi are the angular position and velocity for each data point during a gait cycle, respectively. Such normalization defined the angular positions and velocities between 1 and -1 along both dimensions of the phase plane and minimized individual differences in amplitude and frequency . Phase angles were calculated using the following equation at each time point:The phase portrait for each joint throughout a gait cycle was generated by plotting the normalized angular positions (θ)  . A positce versa . If the Mean absolute relative phase (MARP) was calculated by averaging the absolute values of the ensemble curve points for the tasks :MARP=∑i subject .The deviation phase (DP) was calculated by averaging the standard deviations of the ensemble RPA curve points for the tasks :DP=∑i=1Functionally, a low DP value indicates a stable relationship between the action of two joints or segments for a task and a given subject .The average values of three successful trials were used for statistical analysis. The multivariate analysis of variance (MANOVA) was used to compare the inter-joint coordination parameters between Tai Chi movements or normal walking. The significance level was set at 0.05. All the statistical analyses were performed with SPSS software 20.0 .The flexion and extension angles for the joints changed more frequently in Tai Chi movements as compared with normal walking . As showp < 0.001). During Tai Chi movement, the MARP values of the knee-ankle CRP were significantly smaller than those of normal walking (p = 0.032).This study was the first to examine the inter-joint coordination of older adults while performing Tai Chi movements. The present study results exhibited that the inter-joint coordination between the hip and knee and between the knee and ankle moved nearly in phase or synchronized during Tai Chi movement. Furthermore, low variability of hip-knee and knee-ankle coordination was observed during Tai Chi movements. These findings indicate that the Tai Chi movement has better and more stable inter-joint coordination patterns than normal walking. Therefore, the results of this study supported our hypothesis. The result also agrees with previous studies showing that older adults that regularly practice Tai Chi exhibit better postural stability than those who do not .In addition, the lower-limb joint-angle changes were more frequent during Tai Chi movement as compared with normal walking. During Tai Chi movement, the present study also found that the phase plot of the three joints of the lower extremities appeared in extra closed circles. Based on previous studies, the phase plots in The frequent changes in joint angle indicate that the elderly need to adjust repeatedly, and accurately control the body’s COP and limb position while performing BKTS. This may help regulate mechanoreceptors’ input-output relationship and induce plasticity changes in the central nervous system (such as the increase of synaptic connection strength) . This wiThe patterns of the joint phase plot suggest stable inter-segment coordination for each movement. Therefore, the data on inter-joint coordination must be closely investigated for insights into the overall control of the lower extremity during Tai Chi movements. The dominant joint shifts more frequently during Tai Chi movements, thereby indicating changes in the proximal and distal joint-guided motion patterns. At the beginning of the movement cycle, the proximal joint flexes and acts as an active joint to guide motion. Subsequently, hip-knee and knee-ankle coordination gradually changed from out-phase to in-phase to maintain a COP stability. At the end of the movement cycle, the left leg kicks off, and the lower limb flexes to complete an action similar to crossing an obstacle, thereby making joint coordination and posture more difficult to control.Proximal joints play an important role in postural control, and their dynamic adjustment may increase instability in the body . HoweverHowever, this study encountered three limitations. First, we disregarded the effects on the trunk and upper limbs. Given that Tai Chi is performed with whole-body segment motions, thus these segments might have been affected. Second, the pattern and variability of inter-joint coordination were only examined in the sagittal plane. Given that the sagittal plane is the primary plane of motion during gait, the inter-joint coordination is expected to be robustly controlled and illustrated. Third, all speeds were self-selected. We intended to examine variations in the inter-joint coordination within a range of self-selected paces and minimize interferences in the participants’ performance.Tai Chi movement prompted better adaptability of movement controls and more tightly and stable inter-joint coordination patterns of lower limbs in older adults. This response may reflect a cautious gait pattern, or a functional strategy to help mitigate the risk of falls in older adults. Therefore, Tai Chi exercise may provide a unique way of coordination training, particularly for the coordination between the segments, which may contribute to postural control and thus play a role in preventing falls.


Silicon and heavy metal stress in plants: current knowledge and future prospects” introduces the latest findings in HMs toxicity and the Si-mediated stress tolerance mechanisms in plants so as to promote Si fertilization as a sustainable strategy for mitigating HMs stress and increasing crop productivity. This Research Topic includes four research papers and two review articles.The indiscriminate discharge and consequent accumulation of heavy metals (HMs) from several anthropogenic sources into the environment is a global concern for crop productivity, food security, and human health. During the last few decades, silicon (Si) has been recognized as one of the beneficial elements involved in ameliorating heavy metal toxicities in plants. The large-scale use of Si fertilizers in field crops is also attracting increasing interest. However, our knowledge about Si and HMs stress in plants is still limited. Therefore, the Research Topic “Imran et al., the authors performed the analysis of alternative splicing (AS) under two zinc (Zn) treatment in two different fragrant rice cultivars (Xiangyaxiangzhan and Meixiangzhan-2). RNA-seq analysis revealed dose- and genotype-related AS events in the two rice cultivars. Gene Ontology (GO) analysis showed that these genes were greatly enhanced in a variety of physiological and cellular routes. Moreover, the authors reported that Zn mediates differently the expression of biosynthetic genes of 2-acetyl-1-pyrroline (2-AP). Interestingly, authors also found that genes related to Si, Fe, and other metal-related transport are regulated both at transcriptional and post-transcriptional stages in reaction to Zn. This study highlighted that the biosynthesis pathway of 2-AP and HM transporters can be regulated by epigenetic changes.In the research article by Rabiya et al., a study was conducted to mediate total chromium (Cr) and lead (Pb) stress in soil and Solanum melongena L. This study was done via three bacterial species , CP-silica gel (0.2% chitosan polymerized silica gel), and ZnBc (1.5% zinc-enriched biochar). A 24-h biosorption trial revealed that B3 displayed maximum removal efficiency for Cr and Pb. The results of the pot experiment demonstrated that irrigation with Cr- and Pb-contaminated wastewater significantly reduced plant-growth-related parameters. Nevertheless, the application of ZnBc and CP-silica gel treatments in combination with B3 reduced leaf metal accumulation and ultimately improved phyto-tolerance against oxidative stress due to enhanced antioxidant enzyme activities and leaf total phenolic and protein contents. The study concluded that the use of ZnBc and CP-silica gel in combination with B3 could be used as a sustainable approach to minimize the damage caused by total Pb and Cr coming from wastewater.In another research article by Alsamadany et al., the effect of Si-nanoparticle-doped biochar (SBC) was compared to plain biochar (BC) on improving the salinity and arsenic (As) in quinoa (Chenopodium quinoa). Results showed that growth, yield, and physiological attributes of quinoa were significantly decreased in the mixed stress of salinity and As, which were improved to variable extents both by BC and SBC application. Overall, the SBC showed to be more effective than BC by reducing As uptake and accumulation, thus reducing the oxidative stress load of plants through the overexpression of antioxidant enzymes. Thus, the authors have proposed the amendment of soil with SBC as an efficacious technique to reduce As contamination and enhance food security.In a study by Khan et al. achieved a genome-wide analysis of two transcription factor (TF) families (WRKY and bHLH) in tomato (Solanum lycopersicum) and defined their role against cadmium (Cd) stress. Transcriptomics analysis showed that both genes (bHLH and WRKY) were highly expressed under Cd stress. Results showed that the most bHLHs and WRKYs are DNA-binding proteins, which mediate gene expression negatively and positively. The interaction networks analyses showed that these two families are associated with pathways of stress signaling. In Arabidopsis genome, the ortholog analysis led to classifying bHLH and WRKY orthologs into 11 and 9 clusters, respectively, based on their conserved motif compositions, gene structure, and networks of protein–protein interaction. The authors concluded that identifying HM stress-responsive genes (bHLH and WRKY) in tomato would make available important visions to develop new HM stress-tolerant varieties of tomatoes.In another research article, Li et al., the authors provided a detailed description of the spectroscopy  coupled with advanced machine learning tools for detecting HMs in plants. Recently, spectroscopy techniques application in HMs detection has gained momentum because of its real-time, in situ, and simple action than the traditional chemical analyses techniques. The review summarized the progress of AI-coupled optical imaging spectroscopy techniques application results of plant HM detection over the last decade (2012–2022) and proposed that the challenges of unsatisfactory sensitivity of optimal imaging spectroscopy could be addressed by coupling such analytical and detection methods with advanced AI tools.In the review article by Nikolić et al., the authors summarized the mechanistic understandings of Si interference under variable conditions of iron (Fe) scarcity and copper (Cu) excess. Fe and Cu are known to share similar transport pathways. Consequently, the inadequate concentration of either of the elements may cause disruptions in the availability and uptake of the other, while Si presence in the growth media affects the distribution of the element supplied above or below optimal levels and the distribution of other microelements and their molar ratios. The effectiveness of Si in the stimulation of metal chelators responsible for the mobilization of deficient elements and excess HMs detoxification has been demonstrated. The authors proposed that an investigation into the mechanisms of silicon’s ameliorative effect is valued for decreasing plant mineral stress and enhancing the nutrient composition of crops.In another review article by All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.


The interactions between tumor intrinsic processes and immune checkpoints can mediate immune evasion by cancer cells and responses to immunotherapy. It is, however, challenging to identify functional interactions due to the prohibitively complex molecular landscape of the tumor-immune interfaces. We address this challenge with a statistical analysis framework, immuno-oncology gene interaction maps (ImogiMap). ImogiMap quantifies and statistically validates tumor-immune checkpoint interactions based on their co-associations with immune-associated phenotypes. The outcome is a catalog of tumor-immune checkpoint interaction maps for diverse immune-associated phenotypes. Applications of ImogiMap recapitulate the interaction of SERPINB9 and immune checkpoints with interferon gamma (IFNγ) expression. Our analyses suggest that CD86-CD70 and CD274-CD70 immunoregulatory interactions are significantly associated with IFNγ expression in uterine corpus endometrial carcinoma and basal-like breast cancer, respectively. The open-source ImogiMap software and user-friendly web application will enable future applications of ImogiMap. Such applications may guide the discovery of previously unknown tumor-immune interactions and immunotherapy targets. A statistical analysis framework, termed immuno-oncology gene interaction maps (ImogiMap) provides insights into the tumor immune environment. Combination therapies that target oncogenic processes and immune evasion may induce more durable responses or even curative effects in select cancer types2. It is, however, highly challenging to implement precision therapies involving immunotherapies due to the complexity of the molecular landscape of response predictors.Genomically-targeted therapies and immunotherapies have led to improved patient survival in diverse cancer typesIFNG gene) in the tumor microenvironment4. High expression of SERPINB9 confers resistance to CTLA4 checkpoint inhibition and therefore justifies the therapeutic benefit of co-targeting SERPINB9 and CTLA4. In another example of tumor-immune interactions, DNA repair deficiencies can mediate increased vulnerability to immune checkpoint inhibition through the accumulation of mutation loads leading to neoantigens or activation of the STING-pathway6. Despite such findings, the interactions at the tumor-immune interface, which may inform effective therapies and immune states are relatively unexplored.Identification of biologically relevant tumor-immune interactions that mediate immune evasion by cancer cells may facilitate the discovery of therapeutic targets. For example, the expression of SERPINB9, a potential drug target and a member of the T-cell dysfunction signature genes, is upregulated in tumor cells by interferon gamma , presence of immune checkpoints, and sensitivity to immune effectors 11. Similarly, emerging technologies for spatially resolved omics profiling are highly promising yet suffer from limited sample volume as well as proteomic and transcriptomic coverage13. Therefore, computational methods that can infer tumor-immune interactions from bulk RNA expression data is highly desired and could be useful to identify precision immunotherapies tailored to molecular profiles of tumors of cancer patients. Existing tools such as TIMER and CIBERSORT have been very effective in deciphering the immune signatures in tumors as well as exploring interactions between immune features and the expression of individual genes15. There are, however, no available bioinformatics tools with rigorous statistical validation to investigate the combinatorial tumor-immune interactions that drive immune phenotypes.Single cell sequencing technologies are useful in decoding the heterogeneity of tumor-immune interactions, yet usually limited in sample volumes . Due to the small sample volumes, building single cell data-driven models with high statistical and predictive power remains a hard problem16. The R-package (https://github.com/korkutlab/imogimap) and the web interface (https://bioinformatics.mdanderson.org/apps/imogimap/) may enable rapid adoption of the technology by translational and basic researchers. The method may guide the identification of critical immune checkpoints and inform therapeutic targets within tumors that manifest the tumor-immune interactions.Here, we introduce Immuno-oncology gene interaction Maps (ImogiMap), a bioinformatics method, tool, and web application to automate combinatorial searches for interactions between tumor-associated and immune checkpoint processes. The method generates statistically validated interactions between oncogenic events and immune checkpoints that are co-associated with likely predictors of immunotherapy responses. We have developed the ImogiMap based on the rationale that immune checkpoints that associate with both tumor-related processes and immune phenotypes are more likely to be drivers of immune evasion and therapeutic targets compared to the alternatives without such co-associations. Applications in uterine corpus endometrial carcinoma and basal-like breast cancers (TNBC) nominated tumor-immune interactions, that co-associate with interferon gamma (IFNγ), the key cytokine and effector of antitumor immunity17. TAP genes include but are not necessarily limited to previously defined oncogenes and tumor suppressor genes. ICPs are therapeutically actionable immune checkpoints, for which targeting strategies are in clinical use or trials18. The ICP list is provided within ImogiMap and can be updated by the users  status21, vascularization22, T-cell inflammation signature23, and IFNγ expression16  genes, tumor-associated process (TAP) genes, and immune-associated phenotypes (IAPs) Fig. . TAP gen24. To calculate the combined action score, the patient cohort is stratified into four sub-cohorts  based on TAP and ICP gene co-expression levels. Combined action scores are then quantified based on the IAP levels within the sub-cohorts. Alternatively, the mutation status can be entered to define stratifications . A high combined action score, which is a non-additive deviation of the IAP levels within any one of the sub-cohorts with respect to other sub-cohorts, indicates an interaction between the TAP and the ICP  that quantify IAPs from matched samples. ImogiMap quantifies the tumor-immune interactions with a combined action score between TAP and ICP genes based on their co-association with the IAP of interest. The combined action scores are reminiscent of the synergy scores that are commonly used to define drug-drug interactions based on phenotypic impact of drugs25  that are randomly sampled from the complete cohort. The interaction robustness is quantified as the normalized root mean square deviation of the scores computed from the complete patient cohort vs. partial cohorts. The statistical significance is determined with a Wilcoxon signed-rank test that compares the IAP measurements across the patient sub-cohorts. The null hypothesis for significance is that the median IAP in a “target” sub-cohort and at least one of the remaining three sub-cohorts are sampled from an identical population  and randomly sampled genes (N = 1000) for a particular IAP. When all statistical validation steps are utilized, the resulting p values for significance and specificity can be corrected for multiple hypothesis testing using the Benjamini-Hochberg method. A robust combined action score (not sensitive to moderate changes in data) with high significance  and high specificity  indicates a potentially functional interaction between tumor and immune genes for the IAP of interest.To filter out potential false predictions, each interaction is statistically evaluated for robustness, statistical significance, and specificity. The robustness metric evaluates the stability of the interactions against moderate changes in the exact data configuration. To assess the robustness, we compute the combined action scores using datasets of partial cohorts  and breast cancerIFNG gene expression using the RNA expression profiles of tumors from UCEC patients (source: TCGA)  established as a regulator of TCR and B-Cell Receptor (BCR) signaling30, to be most frequently associated with IFNγ levels through interactions with ICP genes.We identified the significant, specific, and robust interactions that co-associate with GA) Fig. . Our anaed3 Fig. d, e. Togp values from a log-rank statistical test. Among the 23 direct interactions, the co-expression of the CD70 and CD86 pair stood out as most associated with improved progression-free survival in UCEC patients , both molecules promote T-cell activation and immune responses32.Encouraged by these results, we explored other interactions within the network models of tumor-immune interactions in UCEC Fig. . We evalnts Fig. . CD86 anR = 0.55 and 0.64 for CD70 and CD86, respectively) as expected, we asked whether they are differentially associated with specific immune cell types. In the absence of comprehensive single cell omics data, we calculated the partial correlations between CD70 and CD86 genes with diverse immune cell type fractions based on CIBERSORT analysis of RNA expression profiles of tumors from endometrial cancer patients. This is an indirect measure of how each receptor engages with different immune cell types and does not provide definitive proof of their expression sites. We preferred the partial correlation metric as it enables elimination of confounding factors from other random variables while quantifying the direct association between two entities. CD86 is linked with macrophage fractions (both M1 and M2) as well as with regulatory T-cells and activated CD4+ memory T-cells, while CD70 is linked with regulatory T-cells and CD8+ T-cells (Supplementary Table 33 for a detailed discussion of tumor impurity). This argument is also supported by our statistical validations that demonstrated a robust, specific, and significant interaction between the CD70 and CD86 compared to other immune checkpoints.Next, we asked whether the interaction between CD70 and CD86 is due to a co-expression in or co-association with identical immune cell types. Although both immune regulators are associated with overall immune infiltration (R) > 0), significant , and high-ranked combined action scores suggest that expression of CD86 and CD70 co-associate with increased levels of IFNγ in tumors of UCEC patients , robust  , among 44 T-cell dysfunction signature, 29 immune checkpoints, and additional 20 genes that correspond to receptors or ligands of ICPs in CellPhoneDB database, we identified the significant, specific, and robust interactions that co-associate with IFNG gene expression  have been implemented by our and other groups, a few examples being pairwise or partial correlations, database-driven informatics approaches, regression models, ordinary differential equations, stochastic gradient descent for predictive machine learning models and more recently deep learning approachesThe flexibility and statistical rigor of the ImogiMap algorithm may enable generation of hypotheses for the discovery of previously unreported physical or functional interactions. Our current implementation is optimized for immune interactions partly due to the immediate need in the field. ImogiMap, however, is not necessarily limited to tumor-immune interactions, and with its flexibility, it enables different modes of discovery. First, the algorithm can be applied to identify diverse mediators that may together impact a given phenotype (predict interacting genes that associate with a particular phenotype). Execution of ImogiMap with a large gene set against a phenotype of interest may generate a list of interactions that are significantly co-associated with the target phenotype. Second, the algorithm can be used to discover novel interaction partners of a gene, whose impact on a given phenotype is already known . In this case, the ImogiMap can be executed for a pre-defined gene against a large set of candidate genes and the outcomes may guide selection of interactions that mediate a particular phenotype. A major challenge in ImogiMap is selection of likely functional events based on co-associations, which do not dictate causation. This is particularly challenging when baseline, correlative data is used from large patient cohorts as in the case of TCGA datasets. A potential solution to select likely causal interactions is to use time series and perturbation response data. The sequence of expression vs. phenotypic events in time or differential response to perturbations may provide the constraints to establish causality and directionality of interactions. Indeed, the incorporation of such data into ImogiMap is straightforward with minimal design differences. Experimental validation of the predictions, however, is essential to claim a novel discovery regardless of the underlying data modality as well as time and perturbation schemes.42. However, such methods do not capture the higher-level interactions between immune and tumor compartments. As a strong and highly popular tool, TIMER and its web interface provide a way to analysis of immune-tumor interactions14. The TIMER algorithm and web interface enable researchers to explore the immune composition of tumor microenvironments through the use of established immune deconvolution algorithms and compute the correlations between oncogenic alterations with immune cell fractions and phenotypic outcomes . ImogiMap provides a series of advantages over the existing computational approaches. First, the ImogiMap is a statistically rigorous algorithm beyond correlation analyses for the detection of oncogenic alterations and immune checkpoints with an emphasis on therapeutically actionable immune processes. Second, the stratification of patient cohorts based on the co-expression patterns enables not only the detection of interactions but also the establishment of a framework for patient selection in precision oncology applications. Third, the method captures tumor-immune interactions in the context of diverse immune-associated phenotypes such as immune infiltration, immune cell types, IFNγ expression, or inflammation signatures. Fourth, the method incorporates receptor-ligand interactions to assess the tumor-immune interface. Finally, the ImogiMap is highly flexible and enables incorporation of new oncogenic alteration types, immune checkpoints as well as phenotypes.Various immune-oncology bioinformatics and deconvolution algorithms such as CIBERSORT and Kassandra have enabled the estimation of the immune decomposition of tumor ecosystems with increasing accuracyOur findings suggest that ImogiMap generates hypotheses on (and confirms previously reported) relations between tumor-associated processes and immune regulation at a scale not accessible easily by experimental methods. ImogiMap may help basic and translational researchers to discover novel immune-tumor interactions and potential vulnerabilities to combination therapies that target the immune-tumor interactions. The outcomes of ImogiMap analyses may improve the repertoire of actionable ICP targets, and identify patient cohorts that may respond to combination therapies based on their molecular signatures.47. Users have the option to input their own curated list of ICPs based on custom mRNA expression data, proteomics, or quantitative histologic assessments . In the absence of user-provided data, ImogiMap uses API functions in the curatedTCGAData package (Bioconductor), to access TCGA mRNA data. ImogiMap includes a list of 29 immune-associated phenotypes  calculated from scaled IAP values are possible. To demonstrate how the mathematical transformations affect phenotypes, the distribution of phenotypes before and after normalization is depicted in Supplementary Fig. The measurements of IAPs are rescaled and normalized using a logistic sigmoidal function. The scaling facilitates comparisons of phenotypes and resulting combined action scores across different phenotypes. The rescaling is formulated to transform IAPs to a range of . Three separate rescaling functions are formulated based on the initial range of IAP values across samples:The combined action scores between the immune checkpoint and tumor-associated genes are calculated based on the non-linear deviation of the observed IAPs from the expected values for the condition that two genes are independently associated with the IAP. To measure combined action scores that quantify tumor-immune interactions, samples are first stratified into four groups based on median expression levels of two genes, as explained in the main text . First, to calculate a combined action score between two genes, a “baseline” sub-cohort and a target sub-cohort, which carry the presumably least and most co-associated IAP level respectively, are selected among the LL, LH, HL, or HH sub-cohorts. In the absence of a priori assumption on a relationship between the expression of single genes and IAP levels , we analyze all possible configurations such that either the two genes may be activating/deactivating or have opposing relations with the IAP. For each configuration, the relevant baseline and target sub-cohorts with contrasting gene expression levels are identified and a combined action score is calculated as explained below. The configuration which results in the highest combined action score is selected as the metric for the potential combinatorial impact of the two genes on the IAP and is further analyzed.The combined action score is quantified as follows: first, for each sub-cohort, the median deviation of IAP, To reduce noise and ensure that the sign of MN in the target group, is calculated under the condition that each gene acts independently on the IAP. This is achieved through the use of either of the two reference additivity models that are implemented in ImogiMap, the Bliss independence model25 or the Highest Single Agent model24. Under each model In which from Eq.  for the Through statistical assessment of the combined action scores between tumor-associated and immunoregulatory genes, we filter out the scores that are not robust, statistically significant, or specific.R values across diverse interactions with high combined action scores. The least sensitive to the exact data configuration, therefore more robust and reliable scores, have lower “Robustness of interactions” to the underlying data configuration is tested through bootstrapping (1000x) of samples with partial coverage (default is 70% of a complete cohort), re-stratifying patient sub-cohorts based on the bootstrapped data, and calculating the combined action scores. The robustness score is defined as the normalized root-mean-square deviation of combined action scores between complete, p values is reported as the p value of the interaction. The sub-cohort-specific IAP values are compared on boxplots that contain Wilcoxon p values (BH-corrected for multiple hypothesis testing) for group comparisons.“The statistical significance of interactions” is assessed with a Wilcoxon signed-rank sum test comparing the rankings of immune phenotype levels in the target sub-cohort and the remaining three sub-cohorts . In the statistical assessment, the null model is that the immune phenotype levels in the target sub-cohort and at least one of the remaining three sub-cohorts are sampled from the same distribution. The maximum of the three Wilcoxon p value is calculated against the two null models. The highest of the two p values, p value , robust and specific are selected to construct a network model which captures the combinatorial interactions between the tumor-associated and immune genes. In the network models, the edges represent interactions between gene pairs for which a significant, specific, and robust combined action score on IAP exists. Weak interactions are also filtered out based on a user-defined cut-off value. The network model is visualized using the igraph applicationTo interpret interactions from network models, it is important to know potentially relevant ligand-receptor interactions in the tumor microenvironment as they are the key components in mediating cell-cell interactions. To integrate ligand-receptor interactions, we use annotations in the CellPhoneDB database to infer the gene pairs corresponding to protein–protein interactions. Given input gene lists, ImogiMap automatically searches the CellPhoneDB database for potential ligand-receptor pairs and adds them to the original gene lists. Combined action scores are computed for all gene pairs. For each gene pair, the output will include a combined action score, statistical analysis, and whether the gene pair is an inferred ligand-receptor pair in the CellPhoneDB database. The receptor-ligand interactions are annotated on the network models.p values are adjusted for multiple hypothesis testing using the Bonferroni method.In each cohort, the partial correlation between expression of a gene and fraction of immune cell types are calculated using TCGA RNASeq2GeneNorm data and CIBERSORT cell type fractions correspondingly. For each immune cell type, we used R ppcor library to calculate the Spearman partial correlation of the gene-immune cell type while controlling for the rest of the immune cell types. The resulting All statistical analyses are described in detail in the “Statistical evaluation of tumor-immune interactions” section and the main text. To ensure reproducibility, the source code is provided on GitHub (see “Data and code availability section”) and we used publicly available TCGA datasets.Further information on research design is available in the Korkut_Peer Review FileSupplementary InformationReporting Summary


Pityriasis rosea (PR) is a benign, self-limited skin disease characterized by the eruption of multiple erythematous plaques on the trunk and proximal extremities. The proposed etiology links the eruption to the reactivation of human herpesvirus (HHV) 6 and 7 in the skin . HHV-6 vA 2007 Cochrane review  and its P<.001) [Macrolide antibiotics are not recommended for the treatment of PR. Erythromycin was shown in a single clinical trial to significantly reduce the pruritus score compared to a placebo (P<.001) . HoweverP=.04) [P=.004) [P<.001) and resolution of pruritus (P=.02) compared to standard care alone [Acyclovir monotherapy may be useful for the treatment of PR. Compared to a placebo, participants on acyclovir experienced greater resolution of pruritus (P=.04) . Three t[P=.004) . Acyclovre alone .Using the Grading of Recommendations, Assessment, Development, and Evaluations (GRADE) framework, there is moderate quality evidence to support the use of acyclovir monotherapy for the rash and pruritus of PR and low to moderate quality evidence for erythromycin to treat pruritus in PR. A range of acyclovir dosages has been used in studies with unclear evidence based on different regimens . AntivirIn general, the self-limited nature and overall low incidence of PR pose challenges to study designs. Additional high-quality studies are needed to determine if acyclovir or other antivirals are superior to supportive care.


Muscle tension dysphonia (MTD) results from inefficient or ineffective voice production and is the cause of voice and throat complaints in up to 40% of patients presenting with hoarseness. Standard treatment is voice therapy (SLT-VT) delivered by specialist speech therapists in voice disorders (SLT-V). The Complete Vocal Technique (CVT) is a structured, pedagogic method which helps healthy singers and other performers optimise their vocal function enabling them to produce any sound required. The aim of this feasibility study is to investigate whether CVT administered by a trained, non-clinical CVT practitioner (CVT-P) can be applied to patients with MTD before progressing to a pilot randomised control study of CVT voice therapy (CVT-VT) versus SLT-VT.In this feasibility study, we use a mixed-method, single-arm, prospective cohort design. The primary aim is to demonstrate whether CVT-VT can improve the voice and vocal function in patients with MTD in a pilot study using multidimensional assessment methods. Secondary aims are to assess whether (1) a CVT-VT study is feasible to perform; (2) is acceptable to patients, the CVT-P and SLT-VTs; and (3) whether CVT-VT differs from existing SLT-VT techniques. A minimum of 10 consecutive patients with a clinical diagnosis of primary MTD (types I–III) will be recruited over a 6-month period. Up to 6 video sessions of CVT-VT will be delivered by a CVT-P using a video link. The primary outcome will be a change in pre-/post-therapy scores of a self-reported patient questionnaire (Voice Handicap Index (VHI)). Secondary outcomes include changes in throat symptoms , acoustic/electroglottographic and auditory-perceptual measures of voice. Acceptability of the CVT-VT will be assessed prospectively, concurrently and retrospectively both quantitatively and qualitatively. Differences from SLT-VT will be assessed by performing a deductive thematic analysis of CVT-P transcripts of therapy sessions.This feasibility study will provide important data to support whether to proceed with a randomised controlled pilot study focusing on the effectiveness of the intervention compared to standard SLT-VT. Progression criteria will be based on demonstrating a positive outcome in treatment, successful delivery of the pilot study protocol, acceptability to all stakeholders and satisfactory recruitment rates.NCT05365126 Unique Protocol ID: 19ET004). Registered on 06 May 2022.ClinicalTrials.gov website (The online version contains supplementary material available at 10.1186/s40814-023-01317-y. Hoarseness and associated vocal problems (dysphonia) cause both a major impact on quality of life and livelihood for affected patients as well as posing a substantial healthcare burden . ApproxiPrimary MTD is thought to arise from dysregulated or imbalanced laryngeal and para-laryngeal muscle activity due to dysfunctional prefrontal cortical regulation that may interfere with laryngeal motor preparation, initiation and execution combined with heightened input from limbic regions , 5, 6. TTraditional treatment of MTD consists of indirect voice therapy and direct voice therapy  given byThe complete vocal technique (CVT) is pedagogic technique primarily used by singing teachers and vocal coaches to aid singers and actors produce the vocal sound and vocal function that the performer requires  and has Traditionally, voice therapy for patients, and CVT training for performers, has been given face-to-face. However, with the onset of the COVID-19 pandemic, SLT-VT and CVT training have almost entirely been given using telehealth via a video link . There aVocal performers need rapid return to professional vocal function to avoid loss of employment, income and reputation. CVT-VT is used by CVT-P’s for this purpose but there is only anecdotal evidence of its effectiveness in the acute situation. CVT-VT has the potential benefit of providing rapid improvement in MTD patients as it is based on a well-defined, systematic assessment and evaluation protocol . There iPatients with MTD and vocal performers both present with voice impairment and/or reduced vocal capacity due to dysregulated or imbalanced laryngeal and para-laryngeal muscle activity associated with dysfunctional breathing patterns and inefficient use of resonance during phonation. CVT-VT administered by appropriately trained practitioners provides a well-described tool that could potentially be used to restore vocal function in MTD patients. Although CVT-P’s are highly trained, their clientele are largely healthy vocal performers and traditionally they refer clients with unhealthy voices for clinical assessment and treatment. The aim of this feasibility study is to address the following uncertainties before proceeding with a pilot randomised clinical trial of CVT-VT versus SLT-VT: (1) Can CVT-VT applied by a CVT-P improve the voice and vocal function in patients with MTD? (2) Is it feasible to perform a pilot study using CVT-VT administered by a CVT-P using telehealth? (3) Is CVT-VT acceptable to patients, CVT-Ps and SLT-VTs? (4) Does CVT-VT offer a new approach to improving the voice and vocal function compared to traditional SLT-VT methods? Progression criteria will use the traffic light system , 44 and This prospective feasibility study  of CVT-VPatients referred by Primary Care Physicians to the Ear, Nose and Throat (ENT) Department, Queen’s Medical Centre Campus (QMC), Nottingham University Hospitals, with hoarseness will be screened and those with a clinical diagnosis of MTD will be reviewed in a joint voice clinic in the ENT Department, for more specialist assessment. A diagnosis of primary MTD will be made by consensus between an experienced laryngologist and SLT-VT following detailed medical evaluation including laryngostroboscopic examination. Consecutive patients with a confirmed diagnosis of MTD due to 'voice abuse/misuse' Table  who meetAll patients attending the voice clinic in the ENT department at NUH will be assessed by a laryngologist and SLT-VT and have a nasoendoscopic laryngostroboscopic examination prior to recruitment to rule out organic pathology and confirm the pattern of muscle tension dysphonia. Patients identified as having primary MTD seen will have their eligibility checked and given a patient information sheet outlining the study. They will be contacted by phone after their clinic appointment to see if they are willing to consent to taking part. Those interested will be invited to attend a research clinic run by the study laryngologist and SLT-VT. Those who decline will be referred on the standard SLT voice therapy pathway.t = 8), feedback on their experience, and whether goals for treatment were met. Should further therapy be required, this will be arranged in the form of standard SLT-VT.Patient eligibility will be checked, and written consent obtained. Study questionnaires including a goal-setting checklist will be completed. Acoustic and EGG recordings will be made including MPT and the case-report form (CRF) completed. Prior to commencing therapy, all participants will be given indirect voice therapy in the form of a vocal hygiene advice leaflet , specific exercises for appropriate and healthy use of the CVT specific vocal modes that correspond to the desired loudness and quality will be chosen. For low volume or breathy voice symptoms, a more twanged and more intentionally narrowed epilaryngeal space is sought. For hyperfunction symptoms, adjustments in either breath control or releasing of unintentional constriction is sought to counter the hyperfunction and related pressed voice. Moreover, stamina and longevity are addressed by exercises focusing on sustained healthy voice use. CVT techniques and tools addressing the specific needs and impairments of each individual patient will be documented in qualitative notes, allowing for subsequent analysis of the employed intervention strategies. Moreover, patients will be asked to maintain the exercises between therapy sessions, with the CVT-P inquiring about progress between sessions at initiation of every therapy session. The outline of the therapy sessions is listed in Additional file The primary aim of this study is to demonstrate that CVT-VT is beneficial to patients with primary MTD. Secondary aims include (1) whether it is feasible to perform a pilot study using CVT-VT administered by a CVT-P using telehealth; (2) whether CVT-VT is acceptable to patients, the CVT-Ps and SLT-VTs; and (3) whether CVT-VT differs from SLT-VT. The outcomes will provide important evidence for the feasibility of planning a larger pilot randomised controlled trial focusing on the effectiveness of the intervention compared to standard SLT-VT.This question will be addressed using a prospective, non-randomised, single-arm pilot study using a range of multidimensional measures. The study will also test the utility of some of these measures in this patient population by measuring changes in pre- and post-treatment values and provide initial data for the primary outcome measure, to help determine a sample size calculation for a larger trial .Treatment improvement will form one aspect of the evaluation of feasibility to use CVT-VT delivered by a CVT-P to treat patients with muscle tension dysphonia. The objectives are to determine whether CVT-VT is beneficial in the treatment of patients with primary MTD using both patient-rated and physician-measured outcome measures to give a multidimensional assessment of voice and vocal function outcome pre- and post-treatment. Although there is general agreement on some outcome measures such as patient self-evaluation questionnaires, there is less consensus on the auditory-perceptual and objective measures of voice such as acoustic, electroglottographic (EGG) and aerodynamic measures.The patient measures will include two validated disease-specific patient self-evaluation questionnaires the Voice Handicap Index , 51 and Patient-reported outcomes are generally accepted as the most relevant tool for evaluating treatment effectiveness of voice disorders as they may provide a more a more meaningful impact overall of a voice disorder , 54. Of Although voice problems are the most common presenting complaint in patients with MTD, many have associated symptoms of vocal tract discomfort . It is tt = 0, t = 8) using a head-mounted omnidirectional microphone placed approximately 3 cm from the left side of the corner of the mouth. The synchronous EGG signal will be recorded from two electrodes placed on either side of the larynx of the subject and inputted into the laryngograph microprocessor. The signals will be processed using Speech Studio software program (vers. 5.21. Laryngograph® Ltd.) and saved as .wav files. The recordings will then be analysed using the in-built statistical programs and into the computer program Praat [http://www.praat.org/).Pre- and post-treatment acoustic and EGG measurements during phonation provide an objective assessment of the different aspects of the voice and vocal function at specific points in time. There are no universally agreed parameters to measure and so it is common for a range of individual and combined multidimensional parameters to be extracted from both sustained vowels and read text. The quality and standardisation of the recordings is important to avoid measurement error. To that end, the acoustic and EGG measures will be recorded in a treatment room, which exhibits nominal ambient room noise. The recordings will be made at the time of the research clinic appointments  analysis programme in Speech Studio (vers. 5.21. Laryngograph® Ltd.) . The mulThe Praat computer program will be used to calculate CPPS, the AVQI and H1-H2 ratio from the recordings . Unidime2)The sentence “We were away a year ago” (from CAPE-V)In addition, the mean level difference in decibels (dB) between the first and second harmonics (H1-H2) of all voiced segments will be measured for the vocal tasks. H1-H2 is a low-bandwidth measure of spectral tilt that provides an estimate of vocal fold closure during phonation . Less ab3)A passage of read text ‘Arthur the Rat’Other comparative pre- and post-treatment spectral analyses will also be performed using long-term average spectra (LTAS). LTAS is a fast Fourier transform-generated power spectrum of the frequencies comprising a speech sample. Thus, the LTAS is a composite signal representing the spectrum of the glottal source as well as the spectrum or resonant characteristics of the vocal tract. LTAS holds promise as an acoustic index of voice quality . For exaMode speaking fundamental frequency (Fx) (Hz) + coherence %Mode Loudness level (Ax) (dB) + 80% dB range + coherence %Mode contact quotient (Qx) % + 80% contact % range + coherence %Irregularity scores Fx, Ax, Qx (%)◦ 80% minimum and maximum intensity (dB)◦ 80% maximum—minimum frequency range converted to Semitones (Semitone range)Speech range profile (80%)4)Sustained vowels and vowels from speech samples: acoustic formant frequency measures of vocal tract length and formant spaceIt has been argued that connected speech is a better reflection of vocal function compared to sustained vowels . A range5)Happy birthday to youThere is radiographic evidence that individuals with vocal hyperfunction exhibit a significantly higher laryngeal position and reduced hyolaryngeal space with consequent shorter vocal tract lengths (VTL) than individuals with healthy voices –83. RaisThis song has been chosen as it is one of the most widely recognised songs in the English language. However, it is technically quite difficult for non-trained singers as it has a high note, an octave higher,  than the starting note and small intervals that are near each other. Although reaching the top note does depend on the starting note, it is a reasonably good measure of the flexibility of the voice. The proposal is to perform acoustic and EGG recordings whilst singing the four lines. Each line will be analysed for changes in the LTAS spectral slopes pre- and post-treatment.CAPE-VAuditory-perceptual evaluation of voice by expert trained listeners is a subjective judgment on the type and severity of the dysphonic quality present . No audiThe Consensus Perceptual Auditory Evaluation of Voice (CAPE-V) is a standardized clinician-made auditory-perceptual measurement of voice that provides an overall rating of severity as well as discreet ratings of specific vocal parameters including overall severity, roughness, breathiness, strain, pitch and loudness . In this2)CVT speech therapy assessment rating (CVT-STAR)All voice samples will be rated by four experienced SLT-Vs who will undergo a brief refresher training programme in the use of CAPE-V to improve inter-rater reliability , 96. TheDescriptive technical parameters include descriptions of mode of vibration (mainly), the amount of metallic character in the voice, the degree of density in the voice and the chosen vocal mode and vocal mode variation, whether the speaker is within the centre of the chosen vocal mode, to what degree there are vocal effects present, to what degree there are rough vocal effects present and degrees of voice instabilities and degree of strain.Additional speech parameters include rating of sound colour, amount of twang, the speed of speech, the pitch, pitch variation, accentuation/stressing of words, volume and size of the larynx.Parameters for detecting issues include rating the degree of support energy/effort, the degree of economising breath, assessment of inhalation, the opening of the mouth and a final conclusion describing the assessed main issue to be addressed.The CVT speech assessment rates three overall parameters: (1) descriptive technical parameters, (2) additional speech parameters and 3) parameters for detecting issues  with a significant clinical effect (medium effect size of d = 0.79). It was postulated that the improvement was due to improved control and coordination of the respiratory and laryngeal mechanisms associated with reduced physiological effort. It is hypothesised that MPT values will increase post-CVT therapy.MPT is a simple and inexpensive aerodynamic voice parameter for measuring glottal competence and is expressed in seconds . The patThe integrity of the protocol and rationale will be assessed under four main categories: process, resources, management and scientific . The feaAcceptability is a multifaceted construct that reflects the extent to which people delivering or receiving a healthcare intervention consider it to be appropriate, based on anticipated or experiential cognitive and emotional response to the intervention . Patientn = 3) and external to the study site (n = 3). The CVT treatment protocols and assessment methods were reviewed and discussed with CVT teachers at the Complete Vocal Institute in Copenhagen (n = 10). Peer review was obtained by the Innovation Fund Denmark at the point of funding acceptance. The project was also discussed at an NUH Patient & Public Involvement (PPIE) ‘drop-in’ session in December 2021 and with 12 patients attending the Joint Voice clinic at NUH. All gave positive feedback and comments helped inform the final study design and protocol gNumber of voice therapy sessions received by each patient measured using clinical notes at the end of the study and whether patients request additional SLT-VT at the end of the studyAn important measure of acceptability of the study protocol to patients is recruitment and compliance with study protocol  and by using the Rehabilitation Treatment Specification System (RTSS)  applied , with an explicit focus on the detailing the ‘ingredients’ and ‘targets’ of the intervention techniques and exercises from both a CVT-VT and SLT-VT perspective [Transcription of therapy sessions will be performed ad verbumspective , 29, 39.b)Observation of CVT therapy sessions by SLT-VThose patients who agree for their therapy sessions to be recorded will have these recordings made and stored in line with Trust-approved guidelines. The anonymised sessional recordings will then be transcribed and redacted to exclude any personal or identifiable information. The transcripts will be coded using the qualitative research management software NVivo based on the principles of template analysis , a commoAll patients will be asked to give specific consent for observation of their therapy sessions by the experienced study SLT-V. However, not all those who give consent will be observed for practical reasons, but the aim is to observe at least one patient through the 6 weeks of their therapy and sample a further six therapy sessions at different stages of their therapy. The observations will aid the SLT-V in identifying which ‘ingredients’ and ‘targets’ are being used and if they differ from traditional SLT-VT techniques. Further observations regarding the delivery methods, the type of feedback, progression rules and dosing of the ingredients will be made in line with the RTSS framework , 29. An The justification for the sample size is based on the objective of assessing feasibility . In thisDetermining progression criteria is seen as an essential element in assessing the success of a feasibility study . Using aWe will record and report the participant flow according to CONSORT guideline and produce a CONSORT flowchart . As a feThe supervising team and Sponsor  will monitor progress during treatment, consider any adverse effects and use that information to continue or halt the trial. Patients will be offered payment for travel costs for face-to-face research clinic appointments or to attend the outpatient clinic for video-linked therapy sessions if personal equipment is not adequate, but not for participation in the study.Quantitative standard descriptive and inferential statistics methods will be applied to compare pre-and post-therapy measures (median + interquartile ranges with 95% CI). Statistical analyses of audio and EGG recorded data will be performed using Speech Studio (Laryngograph) and the SPSS Statistics package .P values (0.5) and effect sizes calculated for the differences in pre- and post-treatment values using the Wilcoxon rank-sum test. Effect size together with the trial parameter data  will be used to determine the size of sample necessary to carry out a fully powered randomised controlled trial comparing CVT-VT to SLT-VT. For CAPE-V both descriptive statistics, the inter- and intra-rater reliability scores using Cohen’s Kappa will be reported for the individual parameters. For the acoustic and EGG measures descriptive summary statistics and pre- and post-therapy paired-samples t tests will be performed again using Wilcoxon rank-sum tests. The relationships between the results of the VHI, VTDS and MPT will be estimated by means of the Pearson r coefficient. Other non-validated questionnaires will be reported using descriptive statistics. Recruitment and retention data will be analysed using descriptive statistics.VHI, VTDS and MPT will be reported using descriptive statistics (median + interquartile ranges with 95% CI) and the Favourable ethical opinion for conduct has been granted by the East of England - Cambridge South Research Ethics Committee (Reference no. 22/EE/0047).The aim of dissemination will be to inform other speech and language therapists and CVT practitioners of the outcome of this approach of treating patients with MTD. This will be achieved through scientific conference presentations and workshops and feedback obtained to inform SLT-VT acceptability. A paper will be written for a peer-reviewed publication, and the results will be published using CONSORT extension guidelines for pilot and feasibility trials.MTD is a common cause of voice problems but is a heterogeneous group of conditions causing varying degrees of functional limitation. The common feature for type I–III MTD is an imbalance of the three main mechanisms for voice production mainly breath support, laryngeal muscle tension and neck and pharyngeal muscle tension (hyper-constriction) affecting resonance. Traditional methods of treatment include a large range of direct voice therapy methods, often used in combination. Although widely employed successfully by SLT-Vs, many of the methods do not have a high level of evidence to support their use, and overlap in physiological aims. In addition, reducing constriction and producing a normal sounding voice do not always equate to improved voice function in demanding physical environments or social situations. CVT-VT is a well-defined structured approach that enables singers and other professional voice users/performers to produce their voice in a healthy manner for their vocal needs. CVT-VT has not been previously applied to a patient population, and this paper outlines the protocol for a feasibility study. The four aims of this study have been described i.e. (1) to demonstrate whether CVT-VT improves the voice and vocal function in patients with MTD as measured using the primary outcome measure (VHI); (2) whether a CVT-VT study is feasible to perform; (3) whether CVT-VT is acceptable to patients, CVT-Ps and SLT-VTs; and (4) whether CVT-VT offers a new approach to improving the voice and vocal function compared to traditional SLT-VT method. In addition, it will provide preliminary evidence on implementability of CVT-VT as a healthcare intervention by assessing the feasibility, acceptability and of this method as well as help develop criteria for future fidelity assessment . Seven pAdditional file 1. Looking after your larynx advice leaflet.Additional file 2. Outline of therapy sessions.Additional file 3. CVT Speech Therapy Assessment Rating_CVT-STAR.Additional file 4. Summary of patient survey.Additional file 5. Patient/CVT-P goals and feedback questionnaire.


Spatial biology analyses (RareCyte Inc.) reveal mechanisms of actions of BOT, a novel innate-adaptive immune activator. These observations have downstream implications for clinical trial designs using neoadjuvant immunotherapy and potentially sparing patients chemotherapy.In patients with locally advanced cancer without distant metastases, the neoadjuvant setting presents a platform to evaluate new drugs. For mismatch repair proficient/microsatellite stable (pMMR/MSS) colon and rectal cancer, immunotherapy has shown limited efficacy. Herein, we report exceptional responses observed with neoadjuvant botensilimab (BOT), an Fc-enhanced next-generation anti–CTLA-4 antibody, alongside balstilimab  in two patients with pMMR/MSS colon and rectal cancer. The histological pattern of rapid immune response observed (“ The mulAs novel and more efficacious drugs are developed, there is a need to revisit the tumor regression grading (TRG) systems which assess histopathological response. The Mandard-TRG system developed in 1994 in France based on features of regression pattern seen in response to chemoradiotherapy in patients with esophageal cancer, is most commonly used . It rougIt is well known that for mismatch repair proficient/microsatellite stable (pMMR/MSS) colon and rectal cancer, immunotherapy historically has shown limited efficacy . While tinside-out (serosa-to-mucosa) pattern of responses observed with neoadjuvant BOT alongside balstilimab  in patients with pMMR/MSS colon and rectal cancer  tumor regression.We herein report detailed analyses on the pattern of exceptional responses in the first 2 patients  to the BOT/BAL regimen seen with only one dose of BOT and two doses of BAL. Interestingly, we observe a unique histologic pattern of therapeutic response shown in Fig. outside-in’ [outside-in’ pattern of response infers the regression bed with the immune infiltrates typically surrounded the residual tumor foci and abutted normal background lung tissue. Conversely, as colon and rectal cancers develop inwards penetrating deeper layers of the colon wall and spreading to adjacent lymph nodes, the immunotherapy response is typified by an ‘inside-out’ (serosa-to-mucosa) regression pattern. As observed in Figs. near the luminal surface within the tumor center, with dense inflammation surrounding the periphery and comprising most of the grossly identifiable tumor bed   Fig. . The infed) Fig. . A Crohned) Fig. .Residual tumor glands demonstrated evidence of ongoing destruction with incomplete lumens and frequent luminal microabscesses Fig. . The infPatient 1). Figure Patient 2). Of note, we demonstrate intra-tumoral microenvironment heterogeneity in terms of immune response. As such, for the second case, we separated the deeper, inflammatory zone of regression (Tumor Area #1) from the more superficial area with residual tumor (Tumor Area #2) to analyze some of the changes seen in these distinct regions  of 6.4 Mut/Mb, whereas the second tumor was a rectal cancer above the peritoneal reflection (candidate for upfront surgery), KRAS-G12V mutant, and with truncating alterations in APC gene and a TMB of 4.7. Final pathology in both cases was ypT1N0MX. Both cases as noted earlier were advanced and node positive prior to starting treatment ).From a mutational and next generation sequencing standpoint, no molecular alterations appeared to explain the exceptional response that was noted. The first pMMR/MSS tumor was in the ascending colon and was Herein, we provide the first comprehensive histologic characterization and the activity of this specific BOT/BAL regimen in patients with pMMR/MSS colon and rectal cancer in the neoadjuvant setting. Previously, work by Chalabi et al. reported in the initial landmark NICHE-1 clinical trial included patients with both pMMR/MSS and dMMR/MSI-High colon cancers using ipilimumab and nivolumab (IPI/NIVO) . SubsequThus far, the BOT/BAL regimen has shown promising activity in patients with metastatic refractory pMMR/MSS colon and rectal cancers, and especially those without untreated liver metastases . Our worinside-out” since residual disease may be limited to superficial bowel wall layers on the final surgical specimen. This observation for the time being remains hypothesis generating and requires confirmation in larger studies.The purpose of adjuvant therapy is to eradicate any micrometastatic disease remaining. Currently, for advanced colon and rectal cancer patients who undergo surgery and found to be node positive (stage III) or high risk stage II, 3–6 months of adjuvant chemotherapy is recommended . NeoadjuThese preliminary results pave the way for further development of this highly active BOT/BAL regimen for patients with pMMR/MSS colon and rectal cancers, with plans to bring it earlier in the journey of a patient to the neoadjuvant setting. Limitations of this report include the descriptive nature of the findings and no direct comparison with other drugs in a similar cohort. Furthermore, it is possible that the pattern of response may not be unique to BOT/BAL, but generalizable to various immunotherapies given in the neoadjuvant setting. This, however, has not been officially reported with other regimens and trials in this setting. The comprehensive analyses using the spatial testing using the RareCyte Inc. platform adds further to our understanding of the immune microenvironment and how this regimen alters cell type within the tumor-stromal interface.Finally, this work also highlights the limitations of the traditional response assessment systems, not just the mere quantification, as the specific pattern of therapeutic response generated may dictate how best to histologically assess the post-therapeutic specimen. While the “outside-in” pattern is relevant to primary parenchymal tumors as described by Cottrell et al., as well as potentially for metastatic sites, the inside-out (serosa-to-mucosa) pattern characterizes the immune response for primary sites of the luminal cut such as colon or rectum that arise from a polyp and penetrate the deeper layers of the walls of the lumen . It woulGiven the novelty of these findings and first reports of activity of BOT in the neoadjuvant setting we decided to disseminate this albeit in the first 2 exceptional responders since there is interest in this drug/combination in patients with pMMR/MSS colorectal cancers. Many trials are in development in this space for both patients with colon and rectal cancers, and the analyses/observations would be of value for other researchers. The NEST study is rapidly accruing and nearing completion . More analyses on the larger cohort once completed would add further to our understanding of neoadjuvant immunotherapy responses in general, and to the mechanisms of actions of BOT.et al. called NICHE-1 (Neoadjuvant Immune Checkpoint Inhibition and Novel IO Combinations in Early-stage Colon Cancer) [The ‘inside-out’ (serosa-to-mucosa) pattern of response was observed and analyzed in patients with brisk major pathologic responses to the BOT/BAL regimen as part of the investigator-initiated trial ”) . NEST-1  Cancer) . As part Cancer) . PatientTissue immune-microenvironment correlates were assessed at baseline as well as in the surgical specimen. For the latter, RareCyte Inc. (Seattle WA), performed a 13-marker immune-oncology panel that was developed by the company to test the pre- and post-treatment colon and rectal cancer samples on a single paraffin-embedded slide simultaneously at 20X using the Orion instrument (Figs. Supplementary Table 1


The virus-host interaction is a continuous coevolutionary process. A critical role in the coevolution of viruses and hosts is attributed to innate immunity. Several defense mechanisms have been adopted by the host to clear viral infections, and several other mechanisms of immune evasion have been exploited by viruses .This Research Topic highlights the current knowledge on host innate immunity pathways and viral escape and explores the recent advances in antiviral therapeutic strategies.The induction of the antiviral state of infected cells depends on the recognition of viral components by host pattern-recognition receptors, whose downstream signaling results in the production of interferons and cytokines, leading to the activation of adaptive responses .Amurri et al. the role of cGAS/STING axis in the control of RNA virus infections has been exhaustively described. The activation of STING has been demonstrated after infection of different RNA viruses. The arising question is: how can these viruses activate the cGAS/STING axis if they do not elicit any DNA intermediate during their life cycle? An indirect activation of STING might be possible, and it may be linked to diverse causes like mitochondrial stress, and chromatin damage in infected cells  are the canonical sensors for viral RNA, while the cyclic GMP-AMP synthase (cGAS) is the primary sensor for double-strand DNA (dsDNA). Despite the recognition of viral RNA and DNA relies on diverse receptors, several cross-talk mechanisms between RNA and DNA sensing pathways occur. For instance, the cGAS/stimulator of interferons genes (STING) axis, well known for being activated in response to DNA viral infections, has emerged as RNA sensing pathway in recent years. In the review of ed cells . The autFalqui et al. elucidate how ISG15 can induce, prolong, magnify, and steer a specific immune response against the human immunodeficiency virus-1 (HIV-1). An increase in the HIV-1 Env-specific host response occurs after coadministration of DNA vectors codifying for both ISG15 wild type and ISG15 mutant in a heterologous prime-boost combination with a Modified Vaccinia virus Ankara (MVA)-based recombinant vector expressing HIV-1 antigens Env/Gag-Pol-Nef (MVA-B). The authors clearly define the impact of ISG15 on the immunogenicity against HIV-1 antigens in mice, highlighting the importance of ISG15 as an immune adjuvant with potential applicability in therapeutic vaccine approaches against HIV-1.The increasing knowledge of innate immune mechanisms has inspired the current challenge of using vaccine adjuvants as a therapeutic strategy against viruses . The goaWang et al. describe OBI as the persistence of replication-competent viral genome, a covalently closed circular DNA (cccDNA), in the nuclei of infected hepatocytes and/or in the blood of individuals who test negative for HBV surface antigen (HBsAg). The failure of viral clearance is due to the longevity of the cccDNA and the lack of effectiveness of a host immune response against the virus. Once HBV infection is established, the OBI status can persist even for life. The clinical implications of OBI include the risk of liver cirrhosis and hepatocellular carcinoma, and HBV reactivation or spread through liver transplant or blood transfusion. The OBI prevalence in the population is still largely undefined, with the major limitation being the lack of sensitive and standardized diagnostic assays.The lack of a long-term immune system to mount effective immune responses against viruses appears to be critical in the occurrence of Occult hepatitis B virus (HBV) Infection (OBI). Corona et al. propose a multi-target drug approach in the effort to develop novel HIV-1 inhibitors. This strategy allows the multiple inhibition of the two catalytic activities of the reverse transcriptase (RT), both polymerase and Ribonuclease H (RNase H), and, in addition, the integrase (IN), in an allosteric mode. 4 out of 35 indolinone-base multitarget compounds were able to block the HIV-1 replication in the micromolar range, with compound 10a being the most promising derivative for further drug development.The molecular understanding of the viral-host interface is essential in the research of rewarding therapeutic approaches to control the spread of viral infections. Despite the continuous efforts of the scientific community, a cure remains still an elusive goal for many infections. It is the case for HIV-1. Since HIV-1 appearance 42 years ago, more than 30 antiretroviral drugs have been approved, and combination therapies have transformed HIV-1 infection into a manageable chronic disease. However, drug toxicity and resistance are still the major threats to therapy success. Kasabe et al. to the aim of identifying novel inhibitors of Chikungunya virus (CHIKV). So far, no licensed drugs or vaccines are available. 9 out of 14 FDA-approved drugs resulted active in blocking CHIKV replication in vitro. In silico docking analysis suggested that the compounds were potentially able to bind the envelope protein, capsid, and three non-structural proteins. Given the ability of some of these compounds to inhibit also DENV replication, they are very promising for future development and might be used in areas where both viruses are endemic.A drug repurposing approach has been proposed by In conclusion, the Research Topic recaps some of the most recent progresses into innate immunity, viral evasion and drug discovery, whose understanding proves to be useful in the future for the control and perhaps prevention of viral infections.EF: Conceptualization, Writing – original draft, Writing – review & editing. AC: Writing – review & editing.