One of the common modalities used in imaging COVID-19 positive patients is chest radiography (CXR), and serves as a valuable imaging method to diagnose and monitor a patients’ condition. Structured reporting templates are regularly used for the assessment of COVID-19 CXRs and are supported by international radiological societies. This review has investigated the use of structured templates for reporting COVID-19 CXRs.A scoping review was conducted on literature published between 2020 and 2022 using Medline, Embase, Scopus, Web of Science, and manual searches. An essential criterion for the inclusion of the articles was the use of reporting methods employing either a structured quantitative or qualitative reporting method. Thematic analyses of both reporting designs were then undertaken to evaluate utility and implementation.Fifty articles were found with the quantitative reporting method used in 47 articles whilst 3 articles were found employing a qualitative design. Two quantitative reporting tools (Brixia and RALE) were used in 33 studies, with other studies using variations of these methods. Brixia and RALE both use a posteroanterior or supine CXR divided into sections, Brixia with six and RALE with four sections. Each section is scaled numerically depending on the level of infection. The qualitative templates relied on selecting the best descriptor of the presence of COVID-19 radiological appearances. Grey literature from 10 international professional radiology societies were also included in this review. The majority of the radiology societies recommend a qualitative template for reporting COVID-19 CXRs.Most studies employed quantitative reporting methods which contrasted with the structured qualitative reporting template advocated by most radiological societies. The reasons for this are not entirely clear. There is also a lack of research literature on both the implementation of the templates or comparing both template types, indicating that the use of structured radiology reporting types may be an underdeveloped clinical strategy and research methodology.This scoping review is unique in that it has undertaken an examination of the utility of the quantitative and qualitative structured reporting templates for COVID-19 CXRs. Moreover, through this review, the material examined has allowed a comparison of both instruments, clearly showing the favoured style of structured reporting by clinicians. At the time of the database interrogation, there were no studies found had undertaken such examinations of both reporting instruments. Moreover, due to the enduring influence of COVID-19 on global health, this scoping review is timely in examining the most innovative structured reporting tools that could be used in the reporting of COVID-19 CXRs. This report could assist clinicians in decision-making regarding templated COVID-19 reports. The chest radiographs (CXRs) of a number of these patients identified pneumonias exhibiting ground-glass opacities (GGOs). Within early February 2020, the World Health Organisation (WHO) had named the disease as COVID-19, and the virus as a severe acute respiratory syndrome coronavirus (SARS-CoV-2). Additionally, by March 2020, the WHO declared that COVID-19 was a pandemic and stated it was incumbent on all nations to undertake mitigation strategies via key approaches—to detect, protect and treat against the virus and through this, to innovate and learn.On 31 December 2019, the first report of a new respiratory virus emerged from Wuhan, China, where 27 patients were diagnosed with what was initially described as a novel coronavirus. Once within the lungs, COVID-19 subsequently features as air space opacities such as GGO, consolidation and reticular interstitial thickening, which is referred to at times as crazy paving. CT has a higher sensitivity for the detection of GGO but CXRs are useful for the ongoing clinical management of the COVID-19 patient and have clear advantages over CT with respect to more rapid imaging results, availability at the point of care and offer a more affordable and available option than CT, especially in developing countries. GGO is defined as increased pulmonary opacity without obscuring the underlying margins of bronchi and vascular margins, whereas consolidation obscures the appearance of the bronchial and vascular margins. The location of infection is another important consideration as COVID-19 is typically seen in multifocal locations predominantly in bilateral peripheral and basal aspects of the chest.From a radiological perspective, COVID-19 infiltrates the nasopharynx via droplet formation and can quickly enter the lower respiratory system. There are several advocated benefits that reporting templates can offer including a uniform description of COVID-19 infection, a choice of language that is readily understood by a range of referrers, and the use of less ambiguous descriptions. Other advantages are the ease of use with templates organised into subheadings relating to disease or an organ system formatted by the selection from bulleted lists or check boxes.Many international radiology associations responded to the challenges of COVID-19 by developing and recommending specific CXR reporting tools that utilise a structured template that could facilitate reporting in a more standardised way other than traditional free text methods.The purpose of this scoping review was to investigate how such structured reporting templates have been used, or recommended to be used, in the reporting of CXRs in patients with, or suspected of having COVID-19. One of the main inclusion criteria employed for the article selection was in the use of either a qualitative or quantitative design of the reporting template. The selected articles were also examined to discover which conclusions were found in the use of either reporting design to justify their use as a methodology or outcome for patient diagnosis and/or patient care. Further, this review also sought to highlight any gaps that may exist in the literature regarding the use of these structured reporting instruments that may inform future research.A scoping review was conducted to evaluate literature published on the application of structured reporting tools used in the reporting of COVID-19 CXRs. The review was based on the original framework proposed by Arksey and O’Malley.What is the extent in the literature on the use or utility of templates for reporting on patients with suspected or confirmed COVID-19 disease using CXR?What gaps exist in the literature concerning the use of either quantitative or qualitative methods including the research on efficacy or comparison of both reporting techniques.The key research questions employed in this scoping review are as follows:An experienced health librarian assisted in the initial selection of these key broad terms as the starting method of our database interrogation and in the selection of the four most appropriate medical imaging databases to use in the study. We examined original, peer-reviewed articles, and grey literature  published in English between January 2020 until April 2022. Systematic searches were undertaken through the four main medical imaging electronic bibliographic databases of Medline, Scopus, Web of Science and Embase. In conjunction with the electronic searches, periodic hand searches using Google Scholar and the online resources offered by the University of Sydney library were used. The key search words, Boolean operators and search tools are summarised in Research articles needed to satisfy a range of criteria to be included in this review. The reporting template needed to be a format that was a pre-designed document with pre-set options. Moreover, the reporting language type employed in the pre-designed document needed to utilise either a qualitative or quantitative description of disease. Publications earlier than January 2020, articles with no evidence of peer review  and variations on the theme of machine learning where the outcome was to describe a machine learning process rather than a clinical outcome or utility, were all excluded. Data extraction tables were generated and identified the following information: authors, publication date, theme of the study or study purpose, reporting methods, structured reporting of either qualitative or quantitative designs. In addition to the studies that satisfied inclusion criteria, grey literature was included as it yielded important data for later analysis.There were 50 articles examined after inclusion and exclusion criteria were applied. 10 radiological societies’ recommendations in addition to the 50 articles retrieved were also included for analysis. Of the 50 articles, 47 employed a quantitative design whilst the remaining 3 articles used a qualitative method. The results that follow are headed Radiological Societies, Qualitative Structured Reports and Quantitative Structured reports.The criteria for inclusion or exclusion of grey literature underwent the same process that applied to all material found via the web search process  guidelines and sought to determine the diagnostic accuracy of COVID-19 CXRs compared to chest CT using the BSTI template (Appendix A) in examining the health outcomes of their patient cohort. The study was undertaken in a British hospital emergency department and the results of 1198 eligible patients showed that the COVID-19 CXRs (0.56) revealed a poor sensitivity in the detection of COVID-19 compared to CT . There were several described limitations in the research such as the non-inclusion of relevant co-variants of blood markers of infection status that were available in all patients’ files. Further, there was a significant amount of missing clinical data in many patients’ records. Moreover, there was only one reporting radiologist which excluded any inter-rater reliability and a potential for undetected under- or over-reporting of COVID-19. The authors conclusions specifically stated that the use of the CXR to assess COVID-19 status was not statistically correlated with any clinical findings, which included “vital signs, laboratory parameters or 30 day outcomes”. There was however a strong correlation found between radiology findings on CT compared to clinical status.The first of the three qualitative studies included in this review, that of Borakati et al also used the BSTI template guidelines (Appendix A) to study its usefulness in a different international cohort of 30 COVID-19 positive patients. Each patient over a 20-day period underwent portable CXRs and were then classified using the descriptions that fitted the radiological appearances of the BSTI guidelines. The study found that the BSTI descriptors matched their cohort, but suggested an improvement in the descriptors of classic COVID-19 with the inclusion of mid-zonal involvement. The authors hoped the study could serve as a benchmark examination that similar health environments with X-ray facilities could copy as an affordable tool for the assessment of COVID-19 infected patients. There were important study limitations listed by the authors. The cohort size was only 30 patients and there was no access to serial imaging to inspect progression of the disease or to examine variable appearances in any of the patients. Beyond the author’s stated limitations, there were no details given of the number of reporting radiologists or any available statistical data to assess radiologist concordance.The second, a study by Durrani et al who sought to examine the utility of a self-designed reporting template on patients admitted in an emergency department who had a suspected SARS-CoV-2 infection. These results were compared to the RT-PCR test, the gold-standard in assessing SARS-CoV-2 infection. There were two radiologists reviewing all CXRs and blind to the RT-PCR results. The authors employed a self-developed qualitative reporting template defined by using five levels of disease: ‘characteristic pattern’, ‘high suspicion pattern’, ‘indeterminate pattern’, ‘unlikely pattern’ and a ‘normal pattern’ (Appendix B). There were 582 patients’ CXRs reported and the absolute concordance between the two radiologists was 71% (414/582). There was also a Fleiss-Cohen weighted Cohen’s k of 81 . They found that a positive COVID-19 CXR was predictive of a positive RT-PCR and chest radiography can be useful in recognising undiagnosed COVID-19. The reporting template designed by the authors was described as “proof of concept” and the feasibility of the reporting tool, as stated by the authors, should be verified by its use in other radiology facilities, examining other cohorts of COVID-19 patients.The third qualitative article found in this review was that of Yates et alThe qualitative design typically use terms that define the characteristic appearance of COVID-19 infections: consolidation, GGO and crazy paving. Moreover, location is important as COVID-19 is typically seen in bilateral peripheral and basal multifocal locations. The reports also focus on overall descriptions employing terms such as characteristic pattern, high suspicion, likely pattern, indeterminate and classic/probable. One advantage of the qualitative assessment tool is that if there is an agreed lexicon by clinical radiology, then the reporting Radiologist can use the terms confidently and the descriptors in time become more commonly used. There is also a prompt report turn-around which is characteristic of all templated reporting instruments. The major disadvantage is that these terms can be seen by the referrer, who is not familiar with agreed lexicons, as vague, too descriptive and equivocal. and Radiographic Assessment of Lung Oedema (RALE) reporting tools are by far the most common instruments used of the 47 quantitative studies found, and were the reporting tool of choice in 33 studies—70% of all quantitative studies examined.Quantitative structured reporting instruments examined in this review used numerical data or disease descriptions associated with numerical data to describe levels of COVID-19 disease by segmenting the lung fields into regions, lobes, or quadrants and assigning a score or severity index to show the extent of disease. The Brixia The study was undertaken in the Italian city of Brescia (Roman description as Brixia), hence the name. This was one of the earliest published studies examining the prognostic value of using a quantitative scoring system in the assessment of the CXRs of COVID-19 patients. The study employed a numerical grading system by way of an 18-point severity scale that applied to a front-on CXR divided anatomically evenly into six regions. The scoring ranged from 0 with no lung abnormalities, a score of 1 which signified interstitial infiltrates, a score of 2 which signified interstitial and alveolar infiltrates and the highest score of 3 which determined alveolar predominance. Hence, the worst infectious state for a patient would be a totalled six regions scoring alveolar predominance in each, giving a final score of 18. The findings in this study found the Brixia scoring method was an effective tool to monitor the progression of COVID-19 infection, particularly in intensive care.The use of the Brixia scoring system, the most adopted quantitative scoring method used in 40% of all quantitative reporting studies found in this review, was first designed and used in early 2020 where concentrated areas of SARS-CoV-2 outbreaks were initially observed in Italy. The application of RALE to the COVID-19 CXRs is more complex than Brixia. RALE divides the anteroposterior or posterioanterior CXR into four quadrants and uses numerical scores of consolidation (1–4) and density (1–3) x four quadrants. The consolidation in each segment refers to the extent or the percentage of the alveolar opacification in that segment. The density score rates the overall density of that opacification. The density criteria of 1 signified hazy, criteria 2 signified moderate and 3 signified dense. As a result, RALE examines the extent as well of intensity (density) of infection. An example in the use of RALE in a worst-case scenario is outlined in Appendix C.The second most common quantitative structured reporting tool, which comprised 30% of all quantitative studies found in this review, is known as RALE and was initially derived from an assessment tool of pulmonary oedema.The use of the RALE or Brixia systems and other author-constructed quantitative reporting templates are based on three important propositions that underpin all quantitative reporting tools found in this scoping review. Firstly, does the reporting tool have utility, secondly does the reporting tool have a predictive value, and finally does the reporting tool act in a manner to verify other clinical findings and in so doing, act as an independent variable. 415 known COVID-19 patients were examined by use of the RALE reporting tool. The RALE score was found to be efficacious in assessing the degree of COVID-19 infection and assisted treatment management. Another study examining the efficacy of the RALE reporting tool was Orsi et al who examined 155 COVID-19 positive patients. The study conclusion stated that the visual scoring on each CXR was easy and highly reproducible and reflected the clinical severity, assisting patient treatment. The study by Abo-Hedibah et al employed the Brixia method in the examination 325 COVID-19 positive cases. The study concluded by stating that the use of Brixia in the assessment of 325 patients was a reliable method to assess the severity of pulmonary parenchymal disease. They also stated that this reporting tool was particularly accurate in assessing moderate to severe cases of COVID −19. An extensive list of studies that sought to verify the utility of the quantitative reporting tool are found in Appendix D.There were numerous studies in this review that verified the utility of the quantitative reporting tool. For example, in the study by Al-Yousif et al, examined the initial CXR of 340 COVID-19 patients admitted to an emergency setting using the Brixia reporting template to determine if the CXR could be an early predictor of mortality. The study conclusion was that the reporting template was a predictive tool that can identify patients at risk of death as well as the need for ventilatory support. Yasin and Gouda similarly sought to identify the prognostic value of their reporting tool. Their instrument divided each lung into two regions and then four scores applied to each region. A score of 1 equated to ≤25% of infection; a score of 2 equated to 25–50% of infection; a score of 3 equated to 50–75% and a score of 4 equated to >75% of infection. The highest level of infection, >75% of infection in both lungs, scored 8. They found that the reporting instrument was an accurate method of predicting the future severity of SARS-CoV-2 infection. Iftikhar et al reviewed 200 diagnosed COVID-19 patients. The clinical rationale for this study was to determine if the quantitative reporting tool was a predictive risk for hospital admission and eventual intubation/mechanical ventilation hospital disease progressed in time, and if this progression correlated with intubation and risk of death. The study found that the severity score did have predictive value in patient admission and later intubation. An extensive list of studies utilising the predictive method are found in Appendix E.The second feature, the prognostic value of the structured quantitative report, was found in many studies and served as a useful tool to predict patient outcomes. For example, Balbi et al which examined the relationship between rates of COVID-19 infection and adiposity indexes in a cohort of 215 patients. The indexes used were abdominal obesity and body mass index (BMI). The results indicated that increased abdominal obesity levels correlated with a higher rate of COVID-19 infection than those patients with elevated BMI. A study by Rejeki et al looked at the use of plasma convalescent transfusion in reducing COVID-19 infection and found that the Brixia scoring method in conjunction with other laboratory parameters were able to determine the efficacy of the plasma treatment. Christanto et al sought to determine the relationship between diabetes mellitus and COVID-19 in a cohort of 538 patients with both pathologies. Both RALE and Brixia reporting tools were used and found a substantial association between increased hyperglycaemia and lung severity in COVID-19 patients aged 60 years or older.Several quantitative studies used either Brixia or RALE reporting tool as the independent variable in examining the relationship between COVID-19 CXRs and variables of health status. An example is the study by Basilico et alThe anatomy is divided into four or six regions and each assigned a scale of usually 1–3 with increased disease the higher number. The regions are totalled to give a final infection score. The advantages of the quantitative method are its utility as a simple, quick, yet reliable tool and easily understood by any clinician.This review has examined the use of templated radiological reporting tools in the assessment of the CXRs of COVID-19 patients. The qualitative structured reporting methods are recommended by most radiological societies. However, there were only three research studies found via an extensive web search that used a qualitative method, and these studies had variable findings. One study concluded that the use of the CXR to assess COVID-19 status was not statistically correlated with any clinical findings. However, a second found that the qualitative reporting method was a useful tool in the assessment of COVID-19 CXRs, but the authors also noted several limitations. The third study did show a positive conclusion where chest radiography of COVID-19 patients was predictive of a positive RT-PCR and suggested that the CXR could be useful in recognising undiagnosed COVID-19. The difference in the structured quantitative reporting tool used in this instance was designed by the authors themselves which offered more result categories within their reporting template compared to the recommended reporting tools from the various radiology societies.This review has demonstrated that the quantitative structured reporting tools are preferred in studies within clinical environments. The reasons for this could be related to the value of a concise and uniform descriptive language of reporting by correlating disease to numerical scores. The quantitative method also appears to offer a good prognostic assessment of rates of disease along with the likelihood of future intubation and mortality risks. Judicious use of the tool should be considered as early diagnosis can be equivocal. We did not find any articles that compared the utility of structured templates to that of free text style, or indeed any articles that compared radiologist perceptions on the use of the structured reporting templates.In searching and researching the use of structured reporting templates, we acknowledge that there was no comparison of this reporting method to that of free text radiological reporting. Free text style, also known as narrative reporting, is widely used in radiology reporting and has been an essential part of clinical communication in radiology discourse for over a century and will clearly continue to be used.As COVID-19 infection and transmission rates in time decline and eventually COVID retreats from public discourse, it is of significant public health benefit that efficient and speedy diagnostic tools are in place to meet any future re-emerging of this contagion. The history of public health in the past 20 years is replete with life threatening contagions such as Ebola, swine flu, and MERS, each with significant mortality rates. For this reason, it is vital that we are pre-prepared as much as we reasonably can. It is the synergistic action of multiple health strategies, including radiology, that contain illness and hopefully an efficient diagnostic tool not unlike the templated quantitative reporting tool we have developed can play a part in any future return of SARS-CoV-2 or any other significant new respiratory virus.There is limited evidence for the implementation or utility for structured reporting templates, primarily due to the limited research scope where they have been deployed. There is however evidence demonstrated by this review that the quantitative structured style of reporting instrument features more prominently in research articles compared to the alternative qualitative design when reporting CXRs on patients with COVID-19, even though the qualitative structured design is advocated by most known international radiology societies.At this point, it is speculative to determine why the quantitative style is preferred by researchers. It could be that a numerical score is an easier method of disease analysis within health environments where the immediacy of clinical results is a requisite within the persistent presence SARS-CoV-2. The answer could also be found in qualitative report designs where subjective terms such as indeterminate, high suspicion, characteristic or unlikely could be too equivocal for the referring clinician. Moreover, the comprehensive search of four databases did not find any studies that compared structured reporting quantitative to qualitative templates for COVID-19.It is evident that more research is required to not only explore the future use of structured qualitative reporting, but to also examine studies that compare the utility of both reporting methods for COVID-19 CXRs or possibly combine elements of both quantitative and qualitative language within one structured format. The implementation of structured reporting templates also needs to be rigorously assessed for consistency, utility and clarity of reporting quality for all stakeholders including readers and referrers in the reporting of COVID-19 CXRs into the future.


The lymphatic system is composed of a lymphatic vascular network and lymph nodes, lymphoid organs, lymphoid tissue and a colorless fluid called lymph. The lymphatic vasculature is a vital component of our bodies and upholds crucial physiological functions. This intricate network of vessels is responsible for lymph drainage to maintain fluid balance. It is essential for immune cell trafficking to support immune functions and is involved in lipid absorption and transport from the gut. Lymph nodes play a pivotal role in the initiation of immune responses by detecting microbial components and foreign substances present in the lymph. The largest lymphoid organ is the spleen which is involved in immune responses and in filtering the blood. Dysfunction of the lymphatic system can lead to a variety of diseases and conditions, such as lymphedema, cancer metastasis, and inflammation. Recent research has revealed organ-specific functions of the lymphatic system and their implications for health and disease .Lymphatic system: Organ specific functions in health and disease,” brought together a Research Topic of five original research articles and three review articles. These offer valuable insights into the lymphatic field and highlight the recent advances in our understanding of lymphatic system formation and function. The contributing articles in this Research Topic cover a broad range of topics related to the lymphatic system. Focusing on visualization and formation of lymphatic vasculature in different organs including the liver, adipose tissue, spleen, and central nervous system (CNS). Highlighting new mechanisms of filtering in lymph nodes and the significance of fluid pressure in cancer. Finally, the Research Topic includes studies discussing the relationship between gene mutations and fetal oedema and modulation of lymphatic valve number in the hope of new avenues for improving lymphatic vessel function.In a focused exploration of the organ-specific functions of the lymphatic system, the Research Topic “Bobe et al.). The liver, a central organ in metabolism and detoxification, has been underexplored for its role in lymph generation and composition. Bobe and his team employed cutting-edge volumetric imaging to unveil the microarchitecture of hepatic lymphatic vessels. They discovered a unique lymphatic network exclusively located within the portal tracts of the liver, which plays a crucial role in lymph flow. This study provides essential insights into liver physiology and lymphatics, highlighting their interdependence.Volumetric Imaging of Hepatic Lymph Vessels . This study uncovered a surprising link between adipose tissue lymphatic vessels and thermogenesis regulation. Researchers have found that adipose tissue lymphatics are a significant source of the neuropeptide neurotensin, which inhibits thermogenesis. By inducing lymphangiogenesis, they impair thermogenic capacity in adipose tissue, shedding light on the neural and hormonal regulation of body temperature.Adipose Tissue Lymphatics and Thermogenesis  within the lymphatic system. They discovered a critical role for resident macrophages and specialized lymphatic sinus structures in filtering lymph-borne substances. Additionally, their research highlighted the stepwise filtration process orchestrated by lymph nodes in preventing the entry of foreign substances into the bloodstream. The study demonstrated the dynamic and organized nature of lymphatic recirculation and its adaptability during inflammatory responses.Micro- and Macro-Anatomical Frameworks of Lymph Nodes Choi et al. provided a comprehensive overview of the complex interplay between lymphatic vessels, cancer, and tumor fluid pressure, suggesting potential applications of lymphatics in antitumor therapy.The Ambivalent Nature of the Lymphatics-Cancer Relationship These articles highlight the importance of constituting a filtering system for lymphatic recirculation and demonstrate lymphatic vessels’ crucial function in immune surveillance and fluid pressure drainage in the tumor microenvironment.Sugiyama and Hirashima reviewed the relationship between gene mutants causing fetal nuchal edema and various developmental anomalies in mice, shedding light on potential diagnostic and prognostic applications.Fetal Nuchal Edema and Gene Mutations. Ogunsina et al.). Mutations in genes governing lymphatic valve development are associated with congenital lymphedema. Investigated the potential of a FOXO1 inhibitor to stimulate lymphatic valve formation. Their research demonstrated the ability of this inhibitor to upregulate valve-forming genes in lymphatic endothelial cells. Furthermore, in mouse models, the inhibition of FOXO1 led to a significant increase in lymphatic valve numbers, offering a potential avenue to improve lymphatic vessel function.Pharmacological Inhibition of FOXO1 in Lymphatic Valve Growth (These articles shed light on the molecular and cellular mechanisms underlying lymphatic vascular development and explore potential therapeutic strategies for lymphatic-related disorders.In conclusion, the articles on this Research Topic provide a comprehensive overview of the organ-specific functions of the lymphatic system in health and disease. They showcase exciting advances in our understanding of the lymphatic system and highlight the complexity of organotypic lymphatic vascular morphogenesis and function. However, how the underlying molecular and cellular heterogeneity is orchestrated into these different functions is less understood. Specifically, the extent of the interplay between the local microenvironment and lymphatic vessels is needed to achieve functional heterogeneity.Overall, this Research Topic depicted the importance of the lymphatic system in maintaining homeostasis and fighting diseases. It also emphasizes the need for further research into the lymphatic system to develop effective diagnostic and therapeutic approaches for lymphatic-related disorders. We hope that this Research Topic will stimulate further research and discussion on the lymphatic system and its role in human health.


Virtual surgical planning for CAD/CAM mandibular reconstruction by titanium prosthesis was recently reported for resected cases. Even if some advantages are evident, difficulties that may arise for TMJ function after reconstruction originate from prosthesis contamination through oral mucosa dehiscence. In these two cases reported of mandibular reconstruction after resection of ameloblastoma by custom-made CAD/CAM titanium prosthesis, the procedures were aimed to preserve the TMJ glenoid cavity and articular disc avoiding functional problems for hemi-mandibular resections that included the condyle (as in case #1) or with condylar preservation (as in case #2) and avoiding intraoral incisions in both cases. The entire surgical planning and prosthetic fabrication were explained with specifications and the sequence of the surgical procedure. Finite elements analysis (FEA) was performed to check the force distribution and efficacy of the prosthetic device (case 1 with hemi-mandibular resection and rehabilitation). Although successful in these two cases, surgical reconstruction of the mandibular defect after resection by a CAD-CAM custom-made prosthesis still shows some drawbacks and failure risks. Several advantages of this technique and the surgical success in these two cases were presented, but limitations and side effects must be considered when cases are selected. According to the World Health Organization definition, ameloblastoma is a benign intraosseous progressively growing epithelial odontogenic neoplasm characterized by expansion and a tendency for local recurrence if not adequately removed. It represents the second most common odontogenic tumor after odontomas, and it is mostly diagnosed in the fourth and fifth decades. According to the clinical, histological, and radiographic characteristics, it could manifest as a uni-cystic, multi-cystic, peripheral, adenoid, and metastasizing form ,2,3.The treatment strategy for ameloblastoma consists of either resective or non-resective surgical therapy. It is possible, for surgical treatment, a conservative or a radical approach . ConservNon-surgical management of ameloblastoma consists of radiotherapy, chemotherapy, or target therapy and it is addressed to patients medically unstable for surgery or to treat ameloblastic carcinoma and recurrent ameloblastoma after multiple postsurgical recurrences ,6,7.Risks for malignant transformation after radiotherapy have been reported .Ameloblastoma is characterized by a high recurrence rate related to surgical treatment strategy, indeed incidence of recurrence ranges from 55% to 90% following conservative treatment, significantly higher than the incidence rate of 15–25% following more radical treatment. However radical treatment strategy is associated with a significantly higher risk of post-surgical complications, a lower rate of prosthetic rehabilitation, and serious aesthetic and functional impairment ,9.After radical surgery, the restoration of function and acceptable aesthetics represent the primary goals of reconstructive surgical procedures. Several options for reconstruction of the bone defect have been described: vascularized or not vascularized bone grafts, bone chip grafts from the iliac crest and conformed custom-made grids in association with bone morphogenic protein, and finally, distraction osteogenesis ,11,12. IThe pedicled flap with or without a titanium reconstruction plate is the technique most often used for mandibular reconstruction , but sevThis reconstruction technique using CAD/CAM technology has several advantages such as visualization of tumor margins, the definition of surgical margins, fabrication of surgical templates and cutting guides, evaluation of continuity defects, identification of ideal plate dimensions and shape for reconstruction, reduction of surgery time and the fabrication of custom-made prostheses . The maiTo investigate the accuracy and effectiveness of various reconstruction approaches, a finite element analysis (FEA) can be performed .FEA is a valuable tool for stress investigations due to its simplicity. It is a numerical method for breaking down complex geometries into a large number of simple domains (elements) linked by nodes. The tensions and deformations in these simple elements can be evaluated at each node once the geometrical model has been subdivided to evaluate the stress and strain of important structures .Reconstructing mandibular defects with titanium prosthesis using FEA support appears to reduce the risk of screw pull-out and plate fractures .The use of FEA also has several utilities in oral rehabilitation, in fact, in many recent studies FEA was adopted as a method to check the advantages of different implant treatment options for edentulous mandibles from a biomechanical standpoint. In recent reports, maximum stress was detected for cantilever and 45° tilted implants by FEA test of the all-on-four prosthesis . AnalysiRecent studies stated that stiff materials reduced stress on implants; also, short implant adoption could avoid cantilevers and tilted implants in atrophic mandibles ,25,26.By describing these two case reports, we aim to illustrate the procedure we followed for reconstructing a large bone defect after mandibular resection for ameloblastoma treatment, using a 3D custom-made titanium prosthesis, also performing a FEA to check force distribution and efficacy of the prosthetic fabrication, preserving the TMJ glenoid fossa and articular disc to avoid functional problems.Informed consent was obtained from all the patients included in this study and the research was approved by the Ethical committee of the University of Salerno .A 54-year-old female patient came to our attention in October 2019 at the Maxillo-Facial Unit of the University Hospital of Salerno for the management of swelling in the right side of the mandible .The patient referred that she received previous multiple operations for ameloblastoma of the left mandibular side performed in other hospitals, with the first operation in 1977, and the second and third operations for relapses in 1980 and 1999. Clinical documentation was not available, and the patient could not describe the kind of surgical operations received.Clinical examination revealed evident extraoral asymmetry of the lower facial third and an intraoral buccal-lingual expansile lesion in the right mandible. Radiographic examination by Rx panoramic and CT scan showed a wide radiolucent lesion extending from dental elements 4.5 to the entire right mandible ramus involving the right mandibular body and the alveolar nerve canal, with perforations of the cortical bone wall on both buccal and lingual sides .For the radiologic characteristics seen on the CT imaging, a diagnosis of invasive and destructive ameloblastoma was made.The treatment plan included a right-side mandibulectomy with 8 mm safety margins and the reconstruction of the affected portion of the mandible with a fibular free flap for microsurgical reconstruction or microsurgical bone graft from other sites such as the iliac crest or scapula. However, the patient refused every reconstructive surgery that involved a second surgical site and therefore impairment due to microsurgical flap elevation at the donor site. Therefore, these reconstructive hypotheses were discarded, and alternatives were explored. Among these, a bone resection alongside a reconstruction by modern 3D techniques applying a CAD-CAM digitalized and customized titanium prosthesis for precision surgery was the solution that best suited the functional and aesthetic patient’s requests.Adequate maintenance of the midline, occlusion, and mandibular dynamics was all achieved through digital processing of TC cone beam, following the acquisition of 3D images to define the resection lines with safety margins and a CAD-CAM procedure to obtain the cutting guides and a prosthesis with the correct dimensions. After the intervention was simulated and planned on 3D models obtained from the CT scans , mandibuBecause of the expansile presentation of the lesion with subsequent deformity, mirroring of the unaffected side of the mandible was performed, and the general shape of the mandibular prosthesis was delineated from this mirrored image a,b.Using additive manufacturing technology 3D plastic prototypes  were manufactured.After the approval of the prototypes, the mandibular prosthesis was made with SLM (selective laser melting) technology, i.e., with a production process by adding laser-stratified titanium powder instead of extrusion from a single block of titanium, thus obtaining a structure with controlled degree of rigidity. This production feature reduces the so-called stress shielding, which is the altered distribution of loads between the prosthesis and the bone. The prosthesis also presented a trabeculation of the internal structure to lighten the weight and ensure better osteoinduction.The prosthesis shape was manufactured by an accurate procedure able to precisely fit with the symphyseal resection stump and was anchored with 8 bicortical screws in the predrilled holes of the prosthesis, with a wing overlapping the symphysis. Bicortical screw fixation was adopted to avoid tilting inside the wing plate holes . Using CT data, a finite element analysis (FEA) of the implant was performed by bioengineers to identify potential causes of mechanical failure . The maiBecause of the undercuts and the trabeculated hollow structure of the mandibular prosthesis, selective laser melting was adopted for manufacturing concerning technical advantages in comparison to the milling technique. Other biomaterials have not been simulated or considered for manufacturing this kind of device because medical titanium Ti6Al4V ELI grade 5 ASTM F136 realized according to SLM (selective laser melting) technique is the most common biomaterial for skeletal prosthesis according to the current literature.Among metallic biomaterials such as stainless AISI 316L steels and Co–Cr alloys, titanium, and its alloys exhibit the most suitable characteristics for biomedical applications, because of their high biocompatibility, specific strength, and corrosion resistance ,28. Titanium and its alloys are currently adopted in implants such as artificial hip joints and dental roots and in all cases in which implants replace hard tissue. They are required to have high strength and long fatigue life, that is, high fatigue strength. Nowadays, a low Young’s modulus equivalent to that of cortical bone is simultaneously required in order to inhibit bone absorption.The stress/strain FEM analysis was conducted assuming a load similar found in the article by Dahake et al. . In partThe resection and reconstruction procedures were carried out to preserve the integrity of the mucosa, avoiding any perforation and any transmucosal device. The operation was carried out completely by trans-cutaneous access in the submandibular area for better aesthetic results of the surgical scar a.The incision was carried out following a sliding surgical plane technique performing an incision of the cutaneous, subcutaneous, and muscular plane in different sites connected by stepwise dissections to obtain a wider regenerating supply with larger tension distribution and better healing results  . The mandibular prosthesis was planned considering the presence in the mandible of the natural lower dental arch teeth up to 4.4 which would allow an acceptable chewing function.Resection lines were performed following the surgical guide fitted to the mandibular symphyseal bone contour. The fixation screw pre-holes were constructed from the surgical guide data to facilitate the accurate fitting of the CAD-CAM custom-made prosthesis b.Since the main problem that can occur with this kind of reconstruction by customized mandibular prosthesis is dysfunction of the TMJ and the need to implant a prosthesis also for the glenoid fossa, a radical right mandibulectomy comprehensive of the right condylar head involved by the tumor was performed. Because the articular disc was not involved by the tumor c, and coThe functionality of the facial nerve with its fibers was preserved while the inferior alveolar nerve, which was entirely incorporated in the tumor, was sacrificed.The postoperative course was good with limited soft tissue edema and good functional recovery from the very first few days, allowing swallowing of soft foods in the early post-operative time after wound healing and chewing after 2 weeks from the operation.A 33-year-old female patient came to our attention at the Maxillo-Facial Unit of AORN “A. Cardarelli” in Naples  for the management of a jaw swelling in the right mandible side due to a recurrence of ameloblastoma.Radiographic examination by panoramic X-ray and CT scan showed a wide osteolytic lesion with tumor invasion of the mandibular canal and the vascular-nervous bundle, with a pathological mandibular fracture due to the presence of the growing tumor . CT scanAfter the biopsy, the histopathological examination confirmed the diagnosis of ameloblastoma recurrence.The patient refused a reconstruction with the fibula due to possible sequelae to the leg and risk for functional deficits. Therefore, the surgical planning included resection of the mandibular body and ramus preserving condyle and right symphysis region. Surgical resection was planned by virtual reconstruction and CAD-CAM custom-made osteotomy guides. Osteotomy size was planned at the sub-condylar and para-symphyseal right level; reconstruction was planned by a hollow structured titanium prosthesis smooth on the outside, and rough inside, designed and built with a CAD-CAM laser-melting technique from CT data a,b.The design and construction technique of the surgical guides and titanium prosthesis was conducted similarly to those previously described for patient #1.Surgery began with a right cervical skin incision with a curved shape at two fingers from the lower edge of the right mandibular body. A skin with a platysma flap was raised, and the dissection was carried on up to the mandibular external bone cortex with periosteal elevation for the entire bone segment with dental extractions of 44, 45, and 46. Mandibular osteotomies at the planned level  were performed using osteotomy CAD-CAM cutting guides designed by digital custom-made with 3D printing procedure as previously described.Fixation was performed with 13 bicortical titanium screws  inserted in the prosthetic wings precisely fitting the cortical bone stumps. Autologous fat was obtained from the left abdominal subcutaneous region using a cannula, which was grafted after the Lipogems technique treatment to cover the entire mandibular prosthesis surface.In addition, the postoperative course of patient #2 was satisfactory with limited soft tissue edema and good functional recovery from the very first few days, allowing swallowing of soft foods in the early post-operative time after wound healing and chewing just after 2 weeks from the operation. Symmetry was respected and functionality was restored. The surgical wounds had optimal healing, there were no complications due to dehiscence and contamination of the mandibular prosthesis. Follow-up was of one year for each patient free from side effects or complications.Even today, mandible reconstruction after large resections is a challenging procedure despite the notable developments in the surgical methods available. Microvascular reconstruction with a fibula or scapula flap remains the gold standard, although these procedures have some disadvantages such as the complexity and invasiveness of the surgical technique with related risks, particularly in elderly patients, and the need for two surgical teams and prolonged post-operatively care, with a donor site causing morbidity and aesthetic relics, which are unwanted by young patients, especially if female. Prosthetic procedures can lead to suboptimal aesthetic–functional results, with problems for the survival of the implant due to limited bone graft height and the absence of keratinized gingiva.The use of CAD-CAM technology for the fabrication of custom-made surgical guides and titanium prostheses from CT scans represents an alternative in some cases, especially in the areas of craniomaxillofacial trauma, orthognathic surgery, and reconstructive maxillofacial surgery after resection for neoplasms, which are not a candidate to postoperative radiotherapy.In patient #1, condylar resection was needed because of tumor invasion of the condylar structure. To limit post-surgical difficulties after mandibular resection with condylotomy such as the resorption of the glenoid fossa with the displacement of the prosthetic condylar head in the skull base, we preserved the TMJ disc allowing stability in the position of the prosthetic condyle and avoiding the risk of bone resorption of the glenoid cavity. This decision was related to the tumor extension and condylar invasion detected on CT images. From the functional point of view, TMJ and condyle preservation provides paramount advantages for masticatory function and surgical procedures. In cases in which a condylar resection is needed, preservation of other TMJ structures such as the disk, muscles, and ligaments is recommended if tumor-free margins can be guaranteed.To avoid the risks of intraoral exposure of the mandibular reconstructive prosthesis, the operation was entirely conducted transcutaneously without creating any dehiscence in the oral cavity. Furthermore, the analysis of the technical and scientific aspects was verified by bioengineers with finite element analysis (FEA) of the mandibular prosthesis, and two areas of stress concentration were identified on the conjunction margin of the plate with the first lower right screw and the anterior surface of the condylar portion of the plate. Despite the presence of these two points characterized by greater stress, the prosthesis was considered suitable for use as it would have been supported at the condylar level by the residual anatomical portion of the articular disc that we were able to maintain; furthermore, at the level of the plate margin with the screws, the stress was not significant as the bicortical positioning of the screws allowed the absence of their tilting and therefore avoided the stress amplification. Several studies have shown the paramount influence of material quality and stiffness on proper stress reduction of the bone stump and fixation screws, as evidenced by finite element analysis (FEA). In particular, it has been highlighted that stiffer framework materials concentrate more stress in their structure, reducing the magnitude of stress on the prosthetic screw .Regarding patient #2, a lipofilling method was adopted using the Lipogems technique, thus avoiding the risks of intra-oral exposure to the prosthesis.The report of these two cases of mandibular reconstruction after a large resection illustrates the use of CAD/CAM technology in the fabrication of a custom-made titanium prosthesis. Although several advantages were found by using this technique through the surgical success we observed, some limitations or disadvantages in its application must be considered that can be resolved through an accurate selection of the patients.This reconstruction technique offers several benefits, including accurate reconstruction, decreased operative time, limited invasiveness, improved predictability of outcomes, improved patient satisfaction, and decreased complications . It is aIn case of failure, a new prosthesis or microsurgical flap or a combination of a new prosthesis with bone reconstruction by microsurgical flaps or bone chips can be considered ,35,36,37Reconstruction of the mandible after large resections remains a challenging procedure despite advancements in surgical methods. The use of CAD-CAM technology for custom-made surgical guides and titanium prostheses offers several advantages such as accuracy, reduced invasiveness, and improved predictability of outcomes. Our report on two cases of mandibular reconstruction highlights the advantages of this procedure. However, limitations and disadvantages also need to be considered, such as risks for patients who are candidates for postoperative radiotherapy and the risk of peri-implantitis after prosthesis exposure. Careful patient selection and management of potential risks are essential to guarantee optimal outcomes.


Predictive Toxicology forms the backbone of the chemical next-generation risk assessment (NGRA), which integrates NAMs to assure human safety without animal testing  , sometimCairns et al. present an important milestone in implementing MPS in efficient toxicity testing of drugs. The study describes the development and the implementation of a statistical experimental design approach to a bone marrow MPS, where the results demonstrate an optimal approach to the design of MPS experiments that could be generalizable to other systems and scientific questions. This highlights the impact and the applicability of the MPS in drug safety.Tao et al. The paper describes the development of a skin-liver-thyroid (Chip3) that allows to investigate the interactions between “organs” of interest connected via microfluidic circulation and integrates the typical exposure to chemicals. The MPS model incorporates relevant exposure route , metabolism in skin and liver, and the biological effects (thyroid hormones) into a single model. The MPS experiments were compiled with PBPK modeling to derive the safe dermal exposure of a chemical in consumer products.Another implementation of MPS together with physiologically based pharmacokinetic modeling (PBPK) for the chemical safety assessment was reported by Valls-Margarit et al. describe the comparison of different network-based methods to identify candidate genes involved in adverse events and propose an approach to produce consensus prediction to increase the confidence in the target gene prediction. The findings revealed variations in the performance of the assessed tools against the benchmark and their capacity for providing novel insights into the adverse effects mechanism of the drug.Gurjanov et al. report, how the reuse of historical data could contribute to the reduction of animal use. Adequately curated and characterized by stringent statistical analyses the historical data could be used to replace or reduce concurrent control groups using so-called virtual control groups, i.e., modeled control groups based on the collected historical data.in vitro assay readouts. This requires an extrapolation of the in vitro concentration-response, based on an in vitro benchmark concentration, into in vivo dose−response data; quantitative an in vitro-to-in vivo extrapolation (QIVIVE). An understanding of the relevant concentration driving the in vitro toxicity rather than simply using the applied  concentration is an important consideration in this step. PBPK modelling integrates the knowledge on the absorption, distribution, metabolism, and excretion (ADME) of a chemical in the human body or organism and provides a means for this extrapolation. In addition, PBPK models facilitate extrapolations across studies, species, routes, and over various exposure scenarios (Algharably et al. have reported an implementation of QIVIVE for prediction of in vivo prenatal exposure of a chemical leading to developmental neurotoxicity in humans based on in vitro toxicity data and discussing several putative neurodevelopmental toxicity mechanisms. The study discusses developing a maternal-fetal PBPK model to perform QIVIVE in a pregnant women population at 15 weeks of gestation.With an increasing need to incorporate NAMs in chemical risk assessment and the concomitant need to find alternatives to animal testing, quantitative hazard characterization fundamentally relies on the interpretation of the cenarios . AlgharaMcNally and Loizou have justified the refinement and calibration of a human PBPK model of a chemical using in silico, in vitro and human biomonitoring data. The modeling approach demonstrates important implications for the read-across approach, as a part of NAMs for the replacement of animals in chemical safety assessments, to calibrate and validated the developed PBPK model against several data streams from another more data-rich source chemical. The considered read across approach afford more confidence for future evaluations of other similar chemicals.Together the articles of the Research Topic illustrate the advent of truly disruptive technologies complementing and replacing traditional animal testing in toxicology. Arguably, this is part of an ongoing scientific revolution , which p


N = 67) read experimental vignettes describing scenarios of comments involving gender stereotypes while near-infrared spectroscopy recordings were made to measure the hemodynamic changes in the prefrontal cortex. Results found a significant correlation between participants’ perceptions of sexism and brain activation in a brain cluster including the right dorsolateral prefrontal cortex and inferior frontal gyrus. There was a significant gender difference where female participants showed a stronger negative correlation compared to male participants. Future research can expand on these initial findings by looking at subcortical structures involved in emotional processing and gender stereotype application as well as examining cultural differences in perceptions of gender stereotypes and sexism.Sexism is a widespread form of gender discrimination which includes remarks based on gender stereotypes. However, little is known about the neural basis underlying the experience of sexist-related comments and how perceptions of sexism are related to these neural processes. The present study investigated whether perceptions of sexism influence neural processing of receiving sexist-related comments. Participants ( Women experience sexism across a multitude of contexts in their everyday lives including the workplace and schools where a number of studies have highlighted the prevalence of sexist incidents experienced in daily life e.g., ,2). Sexi. Sexi2])Negative emotions such as feelings of anger, upset, weakness and sadness are commonly reported reactions to experiences of sexism ,7,8,9,10A large number of studies has investigated gender differences in neural activation involved in emotional reactivity and emotion regulation in terms of the levels of brain activation as well as the brain regions involved. A meta-analysis found that the right amygdala, dorsomedial PFC and left insula were activated in both sexes for negative emotions where increased brain activation in response to negatively-valenced stimuli was more likely to be observed in subcortical and limbic areas—specifically, the left amygdala and the medial prefrontal cortex (mPFC)—for females . With reAlthough neuroimaging studies have not been conducted on experiences of sexism specifically, we can draw on findings from other relevant fields of study such as prejudice, experiences of discrimination and stereotyping in studying the neural processes involved in experiences of sexist events, since sexism is a form of discrimination based on gender. While little is known about neural correlates involved in sexist-related comments involving gender stereotypes, a significant number of studies have investigated gender stereotype processing. Neuroscientific research into the neural basis of prejudice and stereotypes suggests the interactions of different networks of neural structures (see ). Key brExperiences of discrimination and stereotyping also involve social cognition about the source of discriminatory behaviour. One brain area that has been found to be involved in social cognition and processing social information is the medial PFC (mPFC) ,38,39. TSelf-regulation and cognitive control of social emotions and behaviour such as the inhibition of social stereotypical responses are mainly supported by prefrontal areas. Greater activation in the left dlPFC was observed when responses contrasted with stereotypical gender or racial bias  and duriSpecific gender-oriented stereotypes have been associated with particular words and found to be automatically activated when such words are encountered across different languages such as English, Spanish, Italian and German ,63,64,65Gender differences in the neural correlates of gender stereotyping processes have not been explored in depth as well. In the study by , the finWhile studies have looked at how women respond towards experiencing sexism, neuroimaging studies investigating the neural correlates of responses towards sexist events, specifically sexist-related comments, have not been conducted as far as we are aware. Neuroimaging studies have been conducted on experiences of discrimination such as racial bias and stereotyping in general, but sexism and perceptions of sexist events have not been the focus of these studies despite the prevalence of sexist events in the daily lives of women and the well-established link between experiences of sexism and psychological distress. A majority of previous neuroimaging studies also looked at other-referential gender stereotypes and few studies have looked at self-referential gender stereotypes. Hence, as far as we are aware, this is the first neuroimaging study examining the neural processes involved in experiences of sexist events, specifically sexist-related comments. Examining the relationship between individual perceptions of sexism and neural processes contributes to understanding how the neural responses are associated with the individual’s perceptions and reactions to self-referential, sexist comments, which can help to provide insight into understanding the neural mechanisms underlying the link between experiences of sexism and psychological distress. Hence, the present study aims to investigate the association between perceptions of sexism and neural activation in the prefrontal cortex in response to sexist-related versus non-sexist comments.There are two hypotheses in the present study. Firstly, in line with previous studies on gender stereotypes indicating the role of the inferior frontal gyrus, dlPFC and mPFC activity, we hypothesise that there will be a significant association between the perceptions of sexism and neural activation in these areas. We expect that individuals who perceive and construe sexist elements in comments made by others to a greater extent would experience a greater negative emotional reaction. As a result, we expect that lower dlPFC activity will be associated with higher perceptions of sexism. Secondly, based on previous studies indicating gender differences in gender stereotypical bias, we expect to observe a significant difference between male and female groups in the association between perceptions of sexism and brain activation patterns.age = 22.0, SDage = 2.30) were recruited through (i) a psychology undergraduate course and compensated with course credits and (ii) advertisements and compensated with remuneration. The study was approved by the Psychology Ethics Committee at the Nanyang Technological University (PSY-IRB-2020-007) and written informed consent was obtained from participants before completing the experiment.The participants  which operates using light of wavelength 760–850 nm. This system measures relative oxygenated hemoglobin (HbO) and deoxygenated hemoglobin (Hb) concentrations which indicate cerebral activation and deactivation. fNIRS allows monitoring of local blood oxygenation where greater concentrations of HbO are observed in brain regions with higher activity. fNIRS is based on neurovascular coupling, where cerebral blood flow necessary for brain function is coupled with neuronal activity. Activity in a particular brain region results in increases in cerebral blood flow and increased cerebral oxygenation, which is then detected by the fNIRS (see ) for revPreliminary signal processing was conducted using custom scripts developed based on the pyphysio package on Python . Data prhttps://doi.org/10.21979/N9/C3JRXS (accessed on 18 January 2022).Statistical analyses were conducted using Python (Version 3.9.7). First, the NIRS data were analysed at two levels: within-subject and group-level. A general linear model (GLM) was run for each participant where individual beta coefficients were calculated for each vignette-block on the processed HbO concentrations. Next, the Spearman correlation between the individual-level beta coefficients and the ratings of sexist beliefs was calculated for each subject and cluster. The dataset analysed in this study is available in the open access institutional data repository at the link: According to the first hypothesis, a negative correlation between the perceptions of sexism and neural activation in the PFC areas is expected. To verify this hypothesis, a Wilcoxon signed rank test was used to test the computed Spearman correlation coefficients were significantly negative. Finally, a Kruskal-Wallis test was used to test for significant differences in the Spearman correlation coefficients between the male and female groups. The analysis was replicated for each cluster; Bonferroni correction was applied to correct for multiple comparisons .N in The mean of the correlations between perceptions of sexism and the beta coefficients are summarized in p = 0.001). The correlations were not significant in the other clusters, after Bonferroni correction.The Wilcoxon signed rank test found a significant negative correlation between perceptions of sexism and beta coefficients was observed in the right cluster   . This reThe present study investigated the association between perceptions of sexism and neural activation patterns in response to sexist-related and nonsexist comments. Based on previous studies indicating gender differences in gender stereotype tasks, gender differences in the neural activation patterns were also examined.A significant negative correlation between perceptions of sexism and neural activation in the right cluster was observed. The right cluster includes the dlPFC and parts of the inferior frontal gyrus, encompassing BA08, BA09, BA45, BA46 and BA47. The results support the first hypothesis that lower dlPFC activity is expected to be observed with higher perceptions of sexism. We propose that this may be similar to lower dlPFC activity observed in response to maternal criticism  since boAnother possible explanation for the observed result in the right cluster is the role of right frontal activity in stereotype application and response inhibition. The role of the right IFG in domain-general response inhibition has been proposed to be reflective of efforts towards the inhibition of the influence of stereotypes on behaviour during stereotype judgment tasks . The redA significant gender difference in the correlation between perceptions of sexism and brain activation also emerged, which supported our hypothesis. This aligns with previous studies indicating greater gender stereotypical bias in males e.g., ,72). The. The72])In summary, the findings in the present study provide initial evidence on the association between perceptions of sexism and neural processing of sexist-related content. Findings indicate that perceptions of sexism may be associated with activity in brain areas associated with emotional and semantic processing as well as response inhibition. The gender difference in this association suggests that there may be differences in the neural processing of self-referential gender stereotypes and sexism, which could be related to findings of differences in the relationship between perceived discrimination and psychological well-being in men and women ,90. FutuFirstly, the vignettes in the present study were not an exhaustive representation of gender stereotypes and perception of gender stereotypes may differ across individuals. Cultural differences in the perceptions of sexism also imply that future studies can examine neural processing and responses to sexism across different cultural contexts. For example, the same behaviours may not be recognised as sexual harassment by all women ,92. PercSecondly, the present study made use of the fNIRS and was only able to image cortical structures. Previous studies have observed co-activation between the pars orbitalis division of the inferior frontal gyrus and subcortical structures such as the ipsilateral amygdala . Given tAlthough the experience of sexism is common and gender stereotypes about traits and behaviours are pervasive, how individuals perceive and respond to incidents of sexism is not well understood. The findings in the present study provide initial evidence on the association between perceptions of sexism and neural processing of sexist-related content, suggesting that the perception of self-referential gender stereotypes as sexist may be associated with activity in brain areas associated with emotional and semantic processing as well as response inhibition. The gender difference in this association suggests that there may be differences in the neural processing of self-referential gender stereotypes and sexism, implying that experiences of sexism may be perceived and experienced differently between men and women, possibly affecting both groups differently.


The Regions of Interest Multivariate curve Resolution (ROIMCR) methodology has gained significance for analyzing mass spectrometry data. The new SigSel package improves the ROIMCR methodology by providing a filtering step to reduce computational costs and to identify chemical compounds giving low-intensity signals. SigSel allows the visualization and assessment of ROIMCR results and filters out components resolved as interferences and background noise. This improves the analysis of complex mixtures and facilitates the identification of chemical compounds for statistical or chemometrics analysis.SigSel has been tested using metabolomics samples of mussels exposed to the sulfamethoxazole antibiotic. It begins by analyzing the data according to their charge state, eliminating signals considered background noise, and reducing the size of the datasets. In the ROIMCR analysis, the resolution of 30 ROIMCR components was achieved. After evaluating these components, 24 were ultimately selected explaining 99.05% of the total data variance. From ROIMCR results, chemical annotation is performed using different methods:•Generating a list of signals and reanalyzing them in a data-dependent analysis.•Comparing the ROIMCR resolved mass spectra to those stored in online repositories.•Searching MS signals of chemical compounds in the ROIMCR resolved spectra profiles.  Specifications tableMethod detailsIn recent years, Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) has become a widely used method in analytical studies The ROIMCR methodology is composed of two coupled techniques. The first one involves filtering and compressing the mass spectrometry data using the concept of Regions of Interest (ROI) https://github.com/cplqam/SigSel.git.The workflow of SigSel for the analysis of mass spectrometry datasets in the original instrumental acquisition raw format is composed of 3 steps (detailed below): a) Data transfer, visualization, and filtering based on the charge of the measured MS signals, b) data compression and filtering using the Regions of Interest (ROI) method and Multivariate Curve Resolution (MCR) componentization (ROIMCR method) and c) Extraction of chemical information from ROIMCR results to carry out the annotation and identification of the resolved components from their elution and spectra profiles, as well as the quantitative assessment of the observed concentration changes of these components among the analyzed samples and their posterior statistical, chemometrics and pattern recognition analysis. All the developed functions used in this work are in MATLAB programing language, and they are available in the GitHub repository The raw MS datasets contain both the desired chemical signals and those from the instrumental background, noise, and solvent interferences. In these cases, low-intensity signals of sought-after chemical compounds can sometimes be removed from the data due to the use of a too-high ROI noise threshold. To prevent this loss of chemical information, an additional pre-filtering step based on the charge of the signals can be applied to reduce the ROI intensity threshold applied afterward and achieve an increase in the sensitivity of the whole analysis.The proposed SigSel workflow for selecting mass spectrometry signals based on their charge is illustrated in http://www.mcrals.info), which has been described in more detail in other works [Once the charge-filtered datasets are in the MATLAB computing environment, they are compressed and analyzed using the ROIMCR methodology , and those that can be associated with noise signals rather than chemical compounds  can be eliminated if desired. Signals from components associated with background noise can be removed before further processing.After this initial quality control and selection of the MCR resolved components, the peak areas and heights of their elution profiles can be obtained and submitted to further statistical and chemometric analysis to study the concentration variations of the different chemical compounds among samples. Additionally, the mass spectra of each MCR resolved component can be used to annotate and identify the associated chemical compounds using different strategies to facilitate the chemical compound identification: generating a list of MSROI signals and reanalyzing them in a Data Dependent Analysis (DDA) to determine their fragmentation patterns, comparing the ROIMCR resolved mass spectra to those stored in online repositories and public databases, and searching MS signals of known chemical compounds in the ROIMCR resolved elution and spectra profiles .Fig. 2WoIn summary, SigSel is a useful tool for evaluating and interpreting the results obtained from the ROIMCR method when analyzing mass spectrometry datasets from multiple samples. It allows for more efficient and organized data analysis, leading to reliable conclusions. The workflow of the various options for visualization, evaluation, and data extraction is illustrated in Mytilus galloprovincialis) samples exposed to the antibiotic sulfamethoxazole in two different seasons. The samples were analyzed by LC-HRMS using an LC-LTQ Orbitrap Velos™ spectrometer. Further information can be found in previous work https://github.com/cplqam/SigSel.git.The capabilities of the SigSel software package can be better understood by using an experimental data example from a previous LC-MS metabolomics study of mussel  in the LC analysis of all samples. This method of data matrix augmentation is particularly useful for comparing trends among samples and for subsequent processing (as described below).MCR-ALS model with 30 components was able to explain 99.33% of the total data variance. Increasing the number of components did not reveal any additional information about the chemical constituents in the analyzed samples since they would be related to background noise and explain low data variance. Some of the 30 MCR components identified were not actual chemical constituents of the samples but were related to MS signals from the solvent, background, and instrumental noise The interpretation of the ROIMCR results is a two-step process: first, visualization and quality control of the MCR components, and second, extraction of qualitative and quantitative information to identify the chemical compounds associated with the MCR components and determine their relative concentrations in the analyzed samples. This information is then used for statistical and chemometrics analyses. Following the workflow shown in By following this process, 24 MCR components were ultimately selected out of the initial 30, as they are likely associated with the chemical compounds (metabolites) present in the analyzed samples. These components explained 99.05% of the total data variance. The elution and spectra profiles associated with these MCR components can be utilized for identification, as well as statistical and chemometrics analyses.The peak heights and areas of the elution profiles of the MCR-resolved components were calculated as shown in In this example, those MCR components confirmed to be reliable were identified and checked for their presence in the different analyzed samples and examined in more detail using some optional steps. The m/z values associated with each resolved mass spectrum were used for chemical identification using three different methods. The first method uses a list of retention times, MS1 m/z values, and charge state information from SigSel results to obtain their corresponding MS2 spectra from a posterior DDA analysis A. The seROIMCR methodology has been proposed in omics and environmental studies ,15 to obN.A.Carlos Pérez-López: Conceptualization, Methodology, Software, Writing – original draft. Antoni Ginebreda: Data curation, Writing – review & editing. Damia Barcelo: Supervision, Project administration. Roma Tauler: Data curation, Writing – review & editing.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


The incidence of small ruminant pestivirus infections in Greece remains unknown as they have not been diagnosed in the country since 1974 when the most recent Border Disease Virus (BDV) outbreak was reported. The objective of our study was to explore the possible occurrence of pestiviral infections among sheep and goat farms in Greece and to further determine the variants of major concern. Thus, serum samples were collected from 470 randomly selected animals belonging to 28 different flocks/herds. ELISA on p80 antibody revealed the existence of seropositive animals in four out of the 24 studied sheep flocks, whereas all the goats in the four studied herds were seronegative. Viral RNA and antigens were detected in two sheep out of the four seropositive flocks by RT-PCR and ELISA, respectively. Sequencing and phylogenetic analysis showed that the newly identified Greek variants were closely related to the strains of the BDV-4 genotype. One of the BDV-positive sheep demonstrated the diagnostic profile of a persistently infected (PI) animal, providing additional information regarding the source of the infection. This is the first molecular identification of BDV isolates in Greece. Our findings indicate that BDV infections are likely to remain undiagnosed, highlighting the need for further epidemiological studies and active surveillance programs to determine the prevalence and impact of BDV infections on a countrywide level. Flaviviridae family, the genus Pestivirus includes four main species, namely bovine viral diarrhea virus -1, -2 , classical swine fever virus (CSFV) and border disease virus (BDV) , according to the manufacturer’s instructions for pooled serum samples. Briefly, 28 pooled serum samples were created by mixing 40 μL serum per animal in each pooled sample. Each flock/herd was represented by one pooled sample containing the sera of up to 20 animals. Interpretation of the results suggested that a competition percentage of each pooled sample superior or equal to 65% was indicative of a high probability that PI animals were present within the flock/herd. On the contrary, if the competition percentage was below 65%, there was a low possibility of viral circulation within the flock/herd.The serological detection of pestiviruses in the studied population was conducted using a commercially available ELISA kit . According to the manufacturer’s instructions, an internal positive control (IPC) was included during the nucleic acid purification process in order to ensure the efficient RNA isolation. Viral RNA from the tested sera was purified using the PureLinkTM Viral RNA/DNA Mini Kit .To identify potential asymptotic carriers of pestiviruses, all seronegative individual samples originating from the seropositive flocks/herds were tested using a commercially available real time RT-PCR kit  was able to detect all positive BDV sheep, even though it is recommended for the diagnosis of BVD virus in cattle. Indeed, the aforementioned ELISA kit had 100% concordance with real-time RT-PCR results, indicating the potential to be used as an extra-label diagnostic tool for BDV infections after further validation of its diagnostic performance; for efficient validation, it is necessary to test more samples from antigen-positive animals collected from a higher number of farms across the country and under various circumstances. This finding is contradictory to other studies claiming that ELISA is less sensitive than real-time RT-PCR for detecting BDV-4-infected animals . AlthougIn conclusion, the results of the present study indicate that the circulation of small ruminant pestiviruses in Greek sheep is currently low but high enough to be considered an emerging issue of concern. The detection of specific anti-pestivirus antibodies is currently the most common and feasible approach to investigate the spreading of pestivirus infections since there are no available registered vaccines against the disease. However, PCR can be considered a useful complementary diagnostic tool for the confirmation of the diagnosis and the discrimination of PI animals, as it is a low-cost and minimally laborious technique. The combination of the above techniques could set the base for disease monitoring strategies that should be conducted in the entire ruminant population of Greece to eliminate the risk of the virus spreading. In addition, the infectivity status of imported animals should be taken under special consideration.


The last 18 years have brought an increasing interest in the therapeutic use of perinatal derivatives (PnD). Preclinical studies used to assess the potential of PnD therapy include a broad range of study designs. The COST SPRINT Action (CA17116) aims to provide systematic and comprehensive reviews of preclinical studies for the understanding of the therapeutic potential and mechanisms of PnD in diseases and injuries that benefit from PnD therapy. Here we describe the publication search and data mining, extraction, and synthesis strategies employed to collect and prepare the published data selected for meta-analyses and reviews of the efficacy of PnD therapies for different diseases and injuries. A coordinated effort was made to prepare the data suitable to make statements for the treatment efficacy of the different types of PnD, routes, time points, and frequencies of administration, and the dosage based on clinically relevant effects resulting in clear increase, recovery or amelioration of the specific tissue or organ function. According to recently proposed guidelines, the harmonization of the nomenclature of PnD types will allow for the assessment of the most efficient treatments in various disease models. Experts within the COST SPRINT Action (CA17116), together with external collaborators, are doing the meta-analyses and reviews using the data prepared with the strategies presented here in the relevant disease or research fields. Our final aim is to provide standards to assess the safety and clinical benefit of PnD and to minimize redundancy in the use of animal models following the 3R principles for animal experimentation. Perinatal derivatives (PnD) have been widely recognized as therapeutic tools for tissue engineering and regenerative medicine applications in a broad field of diseases or injuries in the past few years. The term PnD includes birth-associated tissues obtained from term placentas and fetal annexes in their naïve or processed form such as the amniotic and chorionic membranes, chorionic villi, the umbilical cord, the basal plate, and the amniotic fluid, as well as the cells isolated from these tissues and the factors released by them  . Preclinhttps://www.sprint-cost.org/) (accessed on April 2023), funded by COST (European Cooperation in Science and Technology) (https://www.cost.eu/), (accessed on April 2023) is an initiative to bridge the gaps between PnD research and their translation into the clinic. Specifically, the SPRINT working group dedicated to preclinical studies aims to collect and critically analyze preclinical data to provide a clear understanding of the therapeutic potential for specific pathological conditions and the underlying mechanisms in different in vivo models. Systematic and comprehensive reviews of preclinical studies identify research gaps for each animal model and the disease of interest and evaluate therapeutic PnD interventions’ efficacy. SPRINT is focused on PnD applications involving differentiation capabilities into tissue-specific cells or paracrine actions via the release of mediating factors acting on resident and progenitor cells, in line with the body of published benefits in animal models of acute injury and chronic inflammatory diseases.The International Network for Translating Research on Perinatal Derivatives into Therapeutic Approaches  (The COST SPRINT Action CA17116) published a consensus paper on the human placenta tissue and cell nomenclature 7116 publ,4,5,6;  guidelines ,10 was d® database [A systematic literature search of the PubMeddatabase  was perfhttps://www.mendeley.com/) (accessed on February 2022). Eligible studies were selected following a workflow compliant with the PRISMA guidelines, as outlined in The database records identified using the search string were then imported into a Mendeley publication library shared with all COST SPRINT Action members and accessible online or via a locally installed application  (https://jcr.clarivate.com/) (accessed on February 2022), [https://icite.od.nih.gov/) (accessed on February 2022)  [https://app.dimensions.ai/discover/publication) (accessed on December 2020)  [A master relational database  including data fields for bibliometric data, publication metrics, and the studies’ experimental data was established C. A Micry 2022),  were conAmerica) ,14,15,16don, UK) ,18 servehttps://automeris.io/WebPlotDigitizer/) (accessed on February 2022) [The extraction of the experimental data from the selected studies is finished. For each disease or research field, the experimental data from the selected publications was extracted by working group members or external collaborators with the respective expertise C. If appry 2022) . The staSystematic reviews are carried out to assess the treatment efficacy and the clinical benefit of PnD by comparing the outcomes measured in clinical scores in non-treated vs. treated animals, when possible D. All tyIt is the aim of the International Network for Translating Research on Perinatal Derivatives into Therapeutic Approaches  to provide independent and scientifically sound knowledge and guidelines to foster the safe and efficient therapeutic use of PnD. To our knowledge, a systematic review of preclinical studies comparing all types of PnD for a variety of diseases and research fields was not conducted prior to SPRINT initiatives. 3 or anti-tumor and immunomodulatory responses [From this extensive work of literature search and selection and data extraction, several systematic and comprehensive reviews are published already. These publications gathered and analyzed the existing literature on the application of preclinical studies on different fields, namely cutaneous wound healing , ovarianesponses . In mostesponses ,5.https://arriveguidelines.org/) (accessed on February 2023) guidelines [To validate future therapies, animal models are still widely regarded as the gold standard for the evaluation of therapies in diseases with a multifactorial pathology or involving complex organs. The meta-analysis of preclinical studies contributes to the 3R principle and will, therefore, have long-term social and ethical impacts. To further improve the long-term use of future preclinical studies, we recommend filling out the Compliance Questionnaire provided by the Animal Research: Reporting of In Vivo Experiments  is generally well documented, but information on the human donor ethnicity, age, gestational age, etc., is often missing or not well described. While the collection of PnD tissue under an anonymized donation scheme without disclosure of personal data is compliant with the Code of Ethics, the documentation of general patient-related data as stated above would be beneficial for future meta-analyses taking these parameters into account.Obviously, there is a significant amount of variability between diseases for the choice of primary and secondary experimental outcome measures. Within research fields, there is often a broad spectrum of analytical and behavioral tests used to assess the efficacy of the treatments. Well-defined and standardized animal models are required to compare outcome measures between studies. Human diseases and injuries come in various distinct subtypes, while animal models often represent only the pathologies’ primary aspect(s). Preclinical models designed for the best possible representation of the human disease might deviate from the standard models that would allow for an adequate meta-analysis of different treatments. This conflict of interest is challenging to overcome. General statements made for the treatment efficacy of types of PnD, routes, time points and frequencies of administration, and dosages might hold true for standardized models but have to be carefully re-evaluated to find the best treatment for a specific pathology or patient population. Nevertheless, the International Network for Translating Research on Perinatal Derivatives into Therapeutic Approaches  firmly believes that in the future, therapeutic applications of PnD and the knowledge gain of their therapeutic potential and underlying mechanisms will clearly profit from an in-depth, comprehensive, and evidence-based analysis of all data available from basic science, preclinical, and clinical studies.