The COVID-19 pandemic imposed a series of behavioral changes that resulted in increased social isolation and a more sedentary life for many across all age groups, but, above all, for the elderly population who are the most vulnerable to infections and chronic neurodegenerative diseases. Systemic inflammatory responses are known to accelerate neurodegenerative disease progression, which leads to permanent damage, loss of brain function, and the loss of autonomy for many aged people. During the COVID-19 pandemic, a spectrum of inflammatory responses was generated in affected individuals, and it is expected that the elderly patients with chronic neurodegenerative diseases who survived SARSCoV-2 infection, it will be found, sooner or later, that there is a worsening of their neurodegenerative conditions. Using mouse prion disease as a model for chronic neurodegeneration, we review the effects of social isolation, sedentary living, and viral infection on the disease progression with a focus on sickness behavior and on the responses of microglia and astrocytes. Focusing on aging, we discuss the cellular and molecular mechanisms related to immunosenescence in chronic neurodegenerative diseases and how infections may accelerate their progression. The ongoing viral pandemic has imposed behavioral changes resulting in increased social isolation and a more sedentary life, which has affected all age groups . HoweverOlder adults are more vulnerable to infectious diseases  due to iThe decline in physical activity imposed by restriction of outdoor activities and sedentary behaviors  is knownIt is already known that SARS-CoV-2 binds to the receptor for angiotensin-converting enzyme 2 (ACE2) , which iPatients with SARS-CoV-2 show elevated levels of pro-inflammatory cytokines mediated by the dysregulation of the nuclear factor kappa B (NF-κB) signaling pathway  and downThus, we considered it to be of interest to review the potential consequences of the effects of the COVID-related inflammatory response on the immune responses linked to chronic neurodegeneration, associated with central or peripheral virus infections. To that end, we here revisited the influences of social isolation, sedentary life, and central or peripheral infections on mouse prion disease progression, as a proxy for the exacerbated immune response of prion-like chronic neurodegenerative diseases  under siFrom a neuropathological point of view, several parallels have been established between prion diseases , AlzheimAlzheimer’s and Prion’s pathologies share synaptic dysfunctions and axonal trafficking defects  and simiPrions are proteinaceous infectious pathogens, devoid of functional nucleic acids that cause a group of fatal neurodegenerative diseases by self-propagating misfolding protein deposition and an associated inflammatory response . Also knThe mechanisms underlying prion-induced neurodegeneration have been widely investigated . Most ofThe forced and prolonged social isolation caused by the COVID-19 pandemic has aggravated the psychiatric symptoms of older people with cognitive impairments . In factEvidence from both animal models and humans demonstrated the physiological benefits of social interaction . TherefoPrevious studies using environmental manipulations in the triple transgenic mouse model of AD (3xTg-AD) were effective in modifying several behaviors but did not change genetically determined AD-like symptoms .Environmental enrichment (EE) and physical exercise have been used to mimic an active lifestyle in humans and previous findings demonstrated that an active life slows AD progression  and HuntBecause EE and exercise moderate immune responses , and agiTo illustrate the effects of exercise and EE on disease progression after intraperitoneal injection, we selected two hippocampal-dependent tasks: burrowing  and the A systematic review dedicated to the identification of the beneficial effects of physical exercise in AD suggests that aerobic exercises are an effective intervention that can attenuate the neuropsychiatric symptoms as the disease progresses . EvidencChronically activated neuroinflammatory processes in neurodegenerative diseases play a central role in their pathogenesis . BecausePrevious findings in the triple transgenic mouse model of AD, which develops both Aβ plaques and neurofibrillary tangles mimicking the temporal- and regional-specific profile of the human disease, suggested that impairment of the peripheral immune system and neuroimmune communication contribute to premature aging of these mice . SimilarThus, the intense microglial activation in chronic neurodegenerative diseases, under influence of both peripheral and central homeostatic changes, damages healthy neural tissue, and then, in response to the factors secreted by dead or dying neurons, microglial activation is chronically maintained and the associated neuroinflammation leads to progressive self-propagating damage .Microglial activation and neuroinflammation have been shown to be modulated by voluntary exercise and EE , which cPrevious consensus established that oxidative stress, DNA damage, mitochondrial dysfunction, excessive accumulation of misfolded proteins, synaptic impairment, and damage to microRNA (miRNA) processing and inflammation  maybe asAstrocytes can also change their homeostatic phenotypes in response to acute and chronic pathologies, showing reactive subtypes with increased expression of the glial fibrillary acidic protein (GFAP) . In the Astrocyte reactivity in the ME7 prion disease mouse model is influenced by EE and exercise, which decreases neuroinflammation and cell reactivity . This isAstrocyte reactivity by upregulation of the glial fibrillary acidic protein astrocyte reactivity in chronic neurodegenerative diseases is associated with nuclear factor kappa B (NF-κB) activation and remodeling of chromatin with subsequent transcription of proinflammatory genes . SustainAmong the infectious diseases, there has been emerging evidence that infectious agents can be part of the environmental risk factors for the aggravation of neurological disorders . This isper se did not change the number of microglia in CA1, virus infection in prion-diseased mice (at 17 weeks post-injection) induced changes in the number and morphology of microglia. We suggested that virus infection exacerbated microglial inflammatory response in prion-infected mice, thus aggravating chronic neurodegeneration (Previous findings using an intranasal Piry neurotropic virus infection, intrahippocampal injection of ME7 prion strain, or normal brain injection, demonstrated that virus-infected prion-diseased mice exhibited higher microglial morphological reactivity and more severe behavioral outcomes than ME7 prion-diseased mice not infected with virus . Althougneration .SARS-CoV-2 has been found to invade the brain via the olfactory, gustatory, and trigeminal pathways, especially at the early stage of infection . Its neuNeuropathological examination of many areas of the central nervous system in aged patients infected with SARS-CoV-2 who died during the disease revealed signs of neuroinflammation with astrogliosis and microglial activation. Microglial nodules and neuronophagia, most prominent in the brainstem, with hypoxic/ischemic changes in many areas of all examined brains, were also evident . In thisFollowing SARS-CoV-2 respiratory infection, choroid plexus epithelial cells are affected by signals from peripheral inflammation followed by activation of the immune system of the brain, such as differential expression of microglial and astrocytic inflammatory-associated genes, dysregulated homeostasis, and peripheral T-cell neuroinvasion . These sIt has been noted that elderly patients infected with COVID-19, who had episodes of delirium, showed significant hyperactivation of microglia in the hippocampus. Together with the inflammatory lesions of the brainstem and the presence of topographic signs and symptoms, in the absence of specific signs of encephalitis associated with SARS-CoV-2, such features constitute the so-called COVID-19 encephalopathic syndrome . While dIt is, therefore, reasonable to infer that a patient who has survived from COVID-19 encephalopathic syndrome, experiencing or not experiencing long-COVID symptoms, may suffer exacerbated neuroinflammation that will accelerate/aggravate the progression of pre-existing chronic neurodegenerative disease.It is important to highlight, however, that although pathogenic mechanisms of age-related neurodegenerative disorders include the seeded aggregation of disease-specific proteins, as in the prion disease model , the incmiRNAs can regulate innate and adaptive immunity by regulating microglia activation, astrocyte reactivity, and by controlling the egress of peripheral immune cells, such as neutrophils, macrophages, T cells, and B cells . miRNAs Social isolation, sedentary life, and infection are all associated with the restrictions imposed by the COVID-19 pandemic rules and the presence of the virus. In this study, we have revisited the effects of sedentary life and infections on mouse prion disease progression, as a proxy for the exacerbated immune response of prion-like chronic ongoing neurodegenerative diseases. Our previous study with mouse prion disease has demonstrated that these influences contribute to the undesirable aggravation of astrocyte reactivity and microglial activation, which results in more severe behavioral outcomes, and acceleration of disease progression. We anticipate that the SARSCoV-2 infection may similarly potentiate ongoing chronic neurodegenerative disease progression in patients surviving to COVID-19. Our findings, and those of other researchers, have demonstrated the benefits of EE and physical exercise, while emphasizing that an active lifestyle may reduce neuroinflammation, cognitive decline, and behavioral abnormalities and may slow disease progression. Thus, a more physically active lifestyle might also be expected to positively impact on the downstream sequelae associated with SARS-CoV-2 infection.All authors contributed substantially to the conception or design of the study; the acquisition, analysis, or interpretation of data for the study; drafting the study or revising it critically for important intellectual content; or final approval of the version to be published; and agreed to be accountable for all aspects of the study in ensuring that questions related to the accuracy or integrity of any part of the study are appropriately investigated and resolved. CWPD, DB, and DA participated in the data interpretation and writing of the final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Non-climacteric fruits, such as strawberry, grape, raspberry, cherry, citrus, and many others, are characterized by ripening transitions that do not strictly depend on a significant increase in ethylene production and an associated rise in respiration rate. The development and maturation processes of these fruits follows a series of molecular and physiological events that leads to modifications in size, color, texture, flavor, and aroma. Although the ripening process of climacteric fruits has been well-documented, the signals triggering in non-climacteric fruit remains poorly understood. However, the involvement of phytohormones in this process has been demonstrated. The current Research Topic on “Hormonal Regulation of Non-climacteric Fruit Development and Maturation” is a combination of research articles and review articles, providing novel insights and detailed overviews on the current knowledge of different aspects of non-climacteric fruit ripening and the metabolites affected by phytohormones.Citrus sinensis (L.) Osbeck], Romero and Lafuente investigated the implication of ABA in the citrus ripening program, particularly affecting cuticle properties and epicuticular wax metabolism, composition, and morphology. Through a series of morphophysiological, metabolites and molecular analysis, the authors demonstrated that ABA has an influence on cuticle permeability by altering wax metabolism and morphology. This study contributes to a better understanding of ABA's effect on wax and, hence, for future approaches aiming at reducing the susceptibility to detached fruit dehydration.Abscisic Acid (ABA) is considered to play important roles in non-climacteric fruit ripening. This phytohormone has been associated with the improvement of anthocyanins content, especially in strawberries and grapes. However, the effect of ABA in other ripening aspects is still poorly explored in the literature. By taking advantage of the availability of a fruit-specific ABA-deficient mutant named Pinalate from the Navelate orange [Reis et al. show evidence that ethylene does not play a key role in strawberry ripening as it did not change the rate of ripening. However, the authors pointed out specific aspects of fruit development and metabolism that were affected by ethephon, which was dependent on the stage of fruit development, and argued that these effects were imparted by a stress response.Although ethylene has been considered the play master in the ripening of climacteric fruits, its contribution to non-climacteric ripening has been investigated. Through the exogenous application of ethephon and the ethylene-action inhibitor 1-methylcyclopropene (1-MCP) in strawberry attached to the plant, Chervin discussed the contribution of starch accumulation or breakdown for the definition of climacteric and non-climacteric fruits. This intriguing review makes us consider and reconsider the fundamental aspects differentiating these two fruit classes and how much we still do not know about fruit ripening and carbohydrate metabolism. Besides the carbohydrates, the role of polyamines in the phytohormone-dependent induction of ripening at the physiological and molecular levels has been considered and was explored by Gao et al. This review provides a deeper understanding of the metabolism and regulatory roles played by polyamines in climacteric and non-climacteric fruit ripening and pointed out the interplay among these metabolites and phytohormones; signaling molecules, such as H2O2, nitric oxide, and Ca2+; and nitrogen signaling. The repercussions of studies in this area may help put another piece in the puzzle, which is the understanding of ripening.Besides the contribution of phytohormones in the ripening of non-climacteric fruits, the signaling cascade following the induction of phytohormones and the whole metabolic changes necessary toward the induction of this phytohormone-dependent ripening is still poorly characterized. In this regard, Ribalta-Pizarro et al. and Jia et al. Through omics researches it has been possible to observe that the metabolism and signaling pathways during the ripening process are similar to those affected during osmotic stresses. Some authors have argued that osmotic stresses trigger the signaling pathway toward the ripening process. In this context, through several water status, water, and potential osmotic parameters, Jia et al. have demonstrated how the osmotic potential can contribute to fruit ripening initiation in strawberries. The change in fruit water potential is most likely caused by the catabolism of large molecules in receptacle cells and the involvement of osmotin-like proteins (OLPs); this decrease in fruit osmotic potential then triggers the onset of fruit ripening. Moreover, Ribalta-Pizarro et al. showed that small changes in water supply through irrigation could affect grape growth and quality. By studying the whole fruit and tissue-specific content of several phytohormones, notably ABA, ethylene precursor, jasmonic acid, salicylic acid, auxins, and cytokinins, the authors provide a view of how these hormones vary along fruit ripening and how and if they are affected by the reduced water supply. Taken together, the studies of Jia et al. and Ribalta-Pizarro et al. provide evidence for the influence of water status on the induction of fruit ripening mediated by hormones.Finally, this Research Topic brings the studies of Given the increased world population, understanding the major players in the ripening process is necessary to develop novel approaches aiming to improve agricultural practices, food security, and minimize losses during harvest, transport, and storage. We hope that the reader will find a useful reference for the state of the art in the hormonal regulation of the non-climacteric ripening process in this Research Topic.VG wrote the first draft. All authors listed have made substantial contribution and approved the manuscript for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Letrozole, an aromatase inhibitor that blocks estrogen synthesis by inhibiting the final step of the estrogen biosynthetic pathway, has been used in the applications of a wide range of infertility settings. It has been more than 20 years since the initial clinical trial of letrozole for ovulation induction. In light of the accumulating clinical and basic evidence, the efficacy and safety of letrozole have been identified. This mini review focuses on our current knowledge of the applications and mechanisms of letrozole for female infertility and various questions are put forward about how letrozole could be more effectively used. In 1993vo assay . In 2004vo assay . From thin vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles , which can reversibly bind to the rate-limiting enzyme P450 aromatase in estrogen biosynthesis pathway and inhibit the conversion of testosterone to estradiol and androstenedione to estrone . The dow) cycles . Letrozo) cycles , 10. In in vitro, in vivo in animals, and humans  methylene] bis-benzonitrile . It was d humans . Plasma d humans . The extd humans . The majLetrozole inhibits the aromatase activity by more than 99%, and endogenous estrogen synthesis by 97%-99% . The mecLetrozole may be used alone or in conjunction with exogenous FSH for OI, but the optimal dose and regimen of letrozole is not yet clear . The proCC affects the development of endometrial and cervical mucus and often leads to the development of multiple follicles . BesidesThe debates over the merits of letrozole and CC in anovulatory infertility have never stopped. For patients with WHO group II anovulation [WHO group anovulation classification was shown in For ovulatory patients, letrozole is also commonly used to increase their chance of becoming pregnant. Typical diagnoses include mild male factor, endometriosis, pelvic factor, and advanced maternal age , 37. DesThe diagnosis of unexplained infertility is despite intensive evaluation of both male and female partners, the etiology may remain unknown . It is iLetrozole would achieve mono-follicular development in most cycles, thereby may reduce multiple gestation pregnancy and OHSS, with a comparable pregnancy success rate with gonadotropins or CC, may become the first choice of treatment for unexplained infertility .Mechanistically, letrozole administration in the early follicular phase during controlled ovarian stimulation (COS) significantly increased the levels of testosterone and androstenedione in follicular fluid , which iFor normal/high responders, co-treatment with letrozole significantly lower gonadotropin consumption and reduce the incidence of OHSS, the pregnancy outcomes are similar or higher than the other groups –52. AdjuIn 2017, a review in Cochrane included 3599 participants, found no conclusive evidence indicating that letrozole with or without gonadotropins differed from gonadotropins, either in the general population or in poor responders undergoing IVF treatment . And theOHSS is a potentially life-threatening complication of ovarian stimulation during the practice of assisted reproductive technologies (ART). In the rat OHSS model, letrozole decreases the level of VEGF, increases the level of PEDF , the com2 in patients receiving freeze-all embryo strategy during luteal phase, but letrozole did not significantly reduce the rate of severe OHSS compared with the control group  levels, co-administration with letrozole results in reduced incidence of OHSS . Howeverol group . The autysiology . The effThe key to the success of FET is to improve the receptivity of the endometrium and the synchronization of endometrium and embryo development. Although Some studies showed letrozole is superior to natural or hormone-replacement therapy (HRT) in terms of clinical pregnancy, live birth  and misc2 by endometriotic deposits, this makes letrozole an attractive therapy for endometriosis  significantly decreased plasma E2 peak concentration , Natural Science Foundation of Hebei Province (H2019206707 and H2019206674), National key Research and Development program (2018YFC1002104).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Lung ultrasound has become increasingly used in both adult and pediatric populations, allowing the rapid evaluation of many lung and pleura diseases. This popularity is due to several advantages of the method such as the low cost, rapidity, lack of ionizing radiation, availability of bedside and repeatability of the method. These features are even more important after the outbreak of the SARS-CoV-2 pandemic, given the possibility of recognizing through ultrasound the signs of interstitial lung syndrome typical of pneumonia caused by the virus. The purpose of this paper is to review the available evidence of lung ultrasound (LUS) in children and its main applications in pediatric diseases. The management of pediatric lung diseases has always been challenging for clinicians because of the variable clinical manifestations and often overlapping symptoms and signs. Traditionally, chest X-ray (CXR) has played a crucial role in the diagnosis of respiratory diseases. However, the potentially harmful effects of radiation exposure due to CXR reduce its applications, especially in children .In recent years, a new imaging application of sonography has spread in emergency departments and clinical practice: lung ultrasound (LUS) . It is qMoreover, in the era of the coronavirus disease 2019 (COVID-19) pandemic, lung ultrasound has further proven its potential, finding application in early diagnosis, follow-up and management of patients affected by SARS-CoV-2 infection ,8.However, this method has some limitations. In fact, LUS is an operator-dependent examination, and the best performance is mainly obtained with experienced operators trained in pulmonary ultrasound. In addition, ultrasound is only able to detect lesions if directly reaching the pleura, with no normally aerated lung between the pleura and the lesion .The purpose of this paper is to review the available evidence of LUS in children and its main applications in pediatric diseases.Children can be scanned in the upright, supine or lateral decubitus position; furthermore, to obtain major compliance in noncollaborating children, the exam can be performed with a warmed-up gel and the child seated in the caregiver’s lap during breastfeeding or by using age-appropriate distraction techniques in order to minimize anxiety.A linear probe of 10 Mhz is the most commonly used transducer for LUS examination . This tyThe compound should also be turned off because the B-lines disappear when the pleura is insonified from different angles, as done in compound imaging. This will cancel the reverberation artifact, reducing the brightness of B-lines.Each hemithorax is divided into three areas: (1) anterior area, delimited by parasternal and anterior axillary lines; (2) lateral area, within the anterior and posterior axillary lines; and (3) posterior area beyond the posterior axillary line. Every area is subdivided into upper and lower halves.The probe is placed vertically, obliquely and horizontally to the ribs in the anterior, lateral and posterior thorax and moved from one intercostal space to the next, in a caudal direction, from the apices to the costophrenic angles in order to cover the entire lung surface . In all To date, several scoring systems have been proposed to assess the degree of lung involvement in different lung diseases to evaluate the distribution of B-lines, one of the most common ultrasound artifacts. These systems divide the thorax into a different number of zones and assign a score based on the distribution of B-lines in the different explored areas during the scan. Among these, the most frequently used in critical care settings are the 4-zone, 6-zone and 8-zone systems introduced by Platz et al. , the 12-As recently observed , the timThe first layers of the chest consist of subcutaneous tissue and muscles. In longitudinal sections, the ribs appear as curved structures with a posterior acoustic shadow. In axial sections, the intercostal acoustic window is used to study tissues located deep into the skin and muscle . In a noBelow the pleural line, the lung is filled with air. This does not allow the direct visualization of the normal pulmonary parenchyma, but some artifacts and echographic findings remain describable.“A-lines” represent some of these artifacts that can be found in a completely healthy lung  14]. Th. Th14]. Another sonographic artifact is represented by the “Z-lines”. These artifacts are formed by reverberation echoes from irregularities on the lung surface and appear as vertical echogenic lines  originating from the pleura that does not erase the A-lines and do not move with lung sliding  16]. Th. Th16]. Although these artifacts are not present in a normal lung, they may be observed in the infant’s lung in the first 48 h of life because of the possible delay in lung fluid resorption .Until 2020, pediatric studies of lung ultrasound were based on the assumption that the ultrasound appearance of a healthy adult lung was the same as that of a child, and no studies had ever been conducted to describe the appearance of the healthy lung in children, particularly infants. Recently, Buonsenso et al. described the normal ultrasound appearance of the lung in infants during the first 6 months of life by performing an ultrasound at 10 days and 1, 3 and 6 months. The study showed that in the first months of life the healthy infant’s lung is characterized by a B-pattern with multiple vertical artifacts that tend to normalize toward the normal A-pattern typical of the adults at six months of age. No consolidations, pleural line abnormalities or pleural effusion were observed in healthy infants .The main indications for lung and chest wall ultrasound in the pediatric population include:Diagnosis and follow-up of neonatal lung diseases;Confirm antenatal diagnosis of lung malformations;Diagnosis and follow-up of pediatric lung infectious diseases (namely bronchiolitis and pneumonia) and lung complications ;Diagnosis and follow-up of pulmonary edema;Diagnosis of thoracic trauma and early detection of signs of child abuse;Follow-up in children undergoing cardiac surgery;Diaphragm ultrasound.Many lung diseases, such as respiratory distress syndrome, transient tachypnea, pneumonia, atelectasis and pneumothorax, can cause respiratory distress in newborns, especially among premature infants. The differential diagnosis of these diseases is often difficult, but LUS can help to discriminate these diseases ,20.Neonatal respiratory distress syndrome (NRDS) is characterized by a functional and structural immaturity of the lung resulting in respiratory distress that appears at birth. The ultrasound findings of NRDS are represented by coalescent B-lines, diffuse and symmetrically distributed in both lungs. These lines are due to the presence of fluid in the interstitial or alveolar compartment, and generally, their number correlates with the severity of the disease, up to “white lung” in the most severe cases. The pleural line appears irregular, poorly defined and thickened. In addition, small areas of hypoechogenic subpleural consolidation can often be observed, especially in the posterior lung fields  21]..21].The 2019 European consensus guidelines on the management of respiratory distress syndrome assert that lung ultrasound could be a useful tool for clinical decision making, and it seems able to differentiate NRDS from other common respiratory disorders of the newborn, reducing exposure to ionizing radiation . In addiTransient tachypnea of the newborn (TTN), also known as “wet lung,” is caused by a failure in the reabsorption of fluid from the fetal lung. It is typical of term or post-term infants in the case of rapid delivery or cesarean section . InfantsMeconium aspiration syndrome (MAS) refers to a multitude of respiratory symptoms caused by meconium aspiration during delivery. It is more frequent in post-term newborns. The ultrasound findings of MAS are as follows: B-pattern appearance, with coalescent or noncoalescent B-lines, several subpleural consolidations asymmetrically spread on both lungs with few spared areas and in severe cases of MAS a white lung pattern .Congenital malformations of the lung and chest, such as congenital pulmonary airway malformation (CPAM) or congenital diaphragmatic hernia, are usually diagnosed prenatally with ultrasound and with MRI . HoweverThe typical ultrasonographic appearance of CPAM is not yet well defined or univocal, but a large hypoechogenic cystic single lesion, small communicating cystic lesions and consolidations similar to those observed in other lung diseases have been described in several patients . The firAs proposed by Corsini et al., pulmonary ultrasound could contribute to the correct diagnosis in congenital diaphragmatic hernia, even faster than chest X-ray, allowing faster treatment and improving the prognosis . The samThe most frequent respiratory diseases in children are bronchiolitis and community-acquired pneumonia (CAP). The diagnosis of these diseases is mainly clinical, and chest X-ray is recommended only in severe clinical conditions or uncertain diagnoses ,32.Recently, some authors focused on the application of ultrasound in these children, with promising data ,34,35,36In community-acquired pneumonia (CAP), the main ultrasound findings are consolidations that appear as hypoechoic areas with evidence of an air bronchogram within and an ecostructural “liver pattern,” with clear margins (indicating a lobar consolidation) or surrounded by confluent B-lines  6]..6].The liver-like ultrasound appearance, which defines the hepatization of the lung, is consequent to the filling of the alveoli with an inflammatory and purulent material that determines a solid and isoechogenic appearance of the lung. Frequently within the consolidations, it is possible to observe a fine hyperechogenic arborescent pattern that defines the air bronchogram due to the presence of air remaining inside the small bronchioles. The air bronchogram typical of pneumonic consolidations is defined as dynamic due to the possibility of observing the air move back and forth with the breaths, a testament to their patency. This ultrasound sign differs from the static air bronchogram, in which air movement is not observable, typical of atelectasis .Sometimes, detecting large consolidations can be difficult, especially in overweight patients. Recently, Møller-Sørensen et al. described an ultrasound finding that may be helpful in detecting larger consolidations, where a thinner and less bright pleural line can be observed when the underlying lung tissue is consolidated .LUS is a valuable method for the monitoring of lung consolidation in children, avoiding frequent exposure to a massive dose of ionizing radiation ,41,42. WFurthermore, it is possible to detect early complications , making ultrasound an excellent tool for monitoring and follow-up the disease . CurrentIn bronchiolitis, the main ultrasound findings are: areas of subpleural consolidations defined as hypoechogenic areas with parenchymatous appearance and irregular margins, the presence of coalescent B-lines  up to the “white lung” or focal presence of multiple B-lines  and abnormalities of the pleural line  47]. Se. Se47]. The use of LUS for the diagnosis of pneumothorax dates back to 1995 . Since tThe role of LUS in identifying pleural effusion is also well established . In factThe lung abscess appears as a well-demarcated capsular structure surrounding a hypoechoic core without internal vascularity on color Doppler . FurtherPleural empyema refers to an infected, purulent and often loculated pleural effusion due to parenchymal infection that spreads to the pleural cavity. Ultrasound examination of the chest revealed a hypoechoic lesion with complex-septated effusions, passive atelectasis, width uniformity and smooth luminal and outer margins . Color DDue to the ability of ultrasonography to investigate the pleural space, estimate the amount of effusion and describe its features, ultrasonography is not only a useful tool for differentiating pleural effusion from empyema and abscess but can also play an important role in the operative decision to drain or not an effusion or to instill fibrinolytic agents in the pleural space or even to resort to more invasive surgical procedures . AlthougAfter the outbreak of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, the rapid diagnosis of COVID-19 interstitial pneumonia, its management and treatment have become pivotal. In the case of COVID-19 pneumonia, the gold standard for the diagnosis and monitoring of the disease is considered computer tomography (CT) . HoweverSARS-CoV-2 infection in the pediatric population appears to be less severe than in the adult population, and in most cases, the disease is mild or moderate . DespiteTo date, the sonographic appearance of SARS-CoV-2 infection has been best described in adult patients. In adults, COVID-19 pneumonia LUS demonstrates bilateral lung involvement with the typical interstitial syndrome pattern characterized by isolated/multiple or coalescent B-lines, sometimes separated by spared areas, a thinned pleural line with some irregularities, and sometimes pleural effusion and subpleural consolidations . B-linesRecently, much evidence has also encouraged the use of lung ultrasound in the pediatric population by describing the ultrasound findings of SARS-CoV-2 infection in children. In particular, Musolino et al. described in a cohort of 10 pediatric patients ultrasonographic findings similar to those observed in adults, with bilateral lung involvement in 70% of cases, pleural line irregularities in 60% of cases and subpleural consolidations in 10% of cases. In addition, vertical reverberation artifacts were observed in 70% of patients. In contrast, pleural effusion was not observed in any patient . In an eSimilar ultrasonographic findings were reported in a study by Denina et al. on a cohort of eight children. Subpleural consolidations were observed in two cases, whereas confluent B-lines were observed in five of the eight patients . In addiHowever, neither of the two studies cited above demonstrated a correlation between ultrasound appearance and the severity of the disease. However, as suggested by Norbedo et al., LUS could play an important role as a triage tool in children with paucisymptomatic and mild forms of the disease and in the management of patients with a negative nasal swab but with ultrasonographic findings suggestive of disease to recommend precautionary isolation of the patient pending repeat nasal swab .Acute pulmonary edema is characterized by accumulation of the fluid in alveoli and interstitial spaces. Its diagnosis with lung ultrasound is increasingly widespread and useful, especially when a CT scan cannot be used or when a rapid bedside examination is necessary. The artifact B-line was used for the detection of extravascular lung water. Different ultrasound scanning protocols are described to evaluate pulmonary edema ,73,74,75Ultrasound plays a primary role in the management of the polytrauma patient. In fact, because hemorrhage is the first cause of mortality and morbidity in trauma patients, the use of a rapid technique such as ultrasound to investigate the presence of hemorrhage in the pericardial or pleural cavity, as well as in the peritoneal cavity, represents today a milestone in the primary Advanced Trauma Life Support (ATLS) survey .Since the mid-2000s, an extension of the Focused Assessment with Sonography for Trauma (FAST) protocol, E-FAST (Extended-FAST), has been developed, aimed at extending the evaluation, previously limited to the heart and abdominal wall, to the thoracic cavity, to search for pleural effusion and pneumothorax . Thanks In addition, recent evidence has shown that ultrasounds may be a useful tool for the detection of bone fractures, in particular rib fractures . A rib fRecently, several studies have described the role of LUS as a complementary tool for CXR in the perioperative evaluation of children undergoing cardiac surgery ,84. LUS Furthermore, some authors have suggested the role of lung ultrasound in the evaluation of the effects of the incremental PEEP recruitment maneuver in children undergoing cardiac surgery in the reduction of atelectasis, improved lung aeration, oxygenation and respiratory system dynamic compliance for kilogram body weight .Ultrasound can be useful to assess diaphragmatic function. In adult patients, there are many clinical indications for ultrasonography of the diaphragm: diagnosis and monitoring of diaphragmatic paralysis, neuromuscular disorders, guidance for needle electromyography, assessment in chronic diseases, traumatic diaphragm rupture, detection of postoperative complication, difficult weaning, estimating work of breathing and titrating ventilatory support .Recently, different studies have evaluated the application of diaphragmatic ultrasound in pediatric age. In particular, some have authors investigated the predictive value of diaphragm ultrasound for weaning outcomes in critically ill children. In particular, the diaphragmatic parameters with better performance were diaphragmatic thickening fraction and maximum inspiratory pressure .Furthermore, an interesting application is that described by Buonsenso et al. The authors showed diaphragmatic US findings in infants with bronchiolitis and they found a correlation between ultrasound diaphragm parameters  and clinical outcomes .Recently, some technical innovations have been introduced regarding lung ultrasound that could make the method even more informative and simplify its interpretation. Moshavegh et al., for example, have proposed an algorithm for the automatic detection of B-lines able to discriminate between ultrasound images of healthy volunteers and patients undergoing cardiac surgery. This innovative method could in the future be used for a better qualitative and quantitative description of B-lines, useful in patients with interstitial lung disease or pulmonary edema .Ultrasound elastography, previously used mainly for the exploration of the liver or other parenchymatous organs, has also been validated for the examination of the lung. This technique allows evaluating the mechanical characteristics of a tissue that may change in case of disease. To date, some evidence has described the possible role of ultrasound elastography in pathologies such as interstitial lung disease, in the evaluation of solid lesions of the lung and pulmonary edema or pleural effusion . In thisThanks to growing evidence, the semeiotics of the healthy and pathological lung is better characterized, and ultrasound increasingly is used in the diagnosis, management, and follow-up of lung diseases in children and infants. Lung ultrasound is rapidly becoming a valuable tool in the hands of the pediatrician, especially in an emergency department setting. The rapidity, repeatability, low cost and possibility to be performed bedside represent the strengths of this method.The spread of this method could determine many benefits, contributing to better use of the economic resources of the health system and to the more prudent management of children that avoid radiation exposure, during diagnosis and follow-up of lung and thoracic diseases.


Population differentiation is a pervasive process in nature. At present, evolutionary studies on plant population differentiation address key questions by undertaking joint ecological and genetic approaches and employing a combination of molecular and experimental means. In this special issue, we gathered a collection of papers dealing with various ecological and genetic aspects of population differentiation in plants. In particular, this special issue encompasses eight research articles and two reviews covering a wide array of worldwide environments, plant functional types, genetic and genomic approaches, and common garden experiments to quantify molecular and/or quantitative trait differentiation in plant populations. Overall, this special issue stresses the validity of traditional evolutionary studies focused on plant populations, whilst emphasizing the integration of classical biological disciplines and state-of-the-art molecular techniques into a unique toolkit for evolutionary plant research. Several ecological and genetic processes acting in concert account for population differentiation, which is a pervasive process in nature that may eventually lead to speciation. In this special issue, we gathered a collection of papers dealing with various ecological and genetic aspects of population differentiation in plants covering a wide array of worldwide environments, plant functional types, genetic and genomic approaches, and common garden experiments to quantify molecular and/or quantitative trait differentiation in plant populations. Mutation, anisotropic migration, random genetic drift and natural selection shape among- and within-population patterns of genetic diversity at different spatial and temporal scales, which steadily promote population differentiation, and in the end, may lead to speciation. This well-accepted statement identifies the major processes underlying evolutionary change, which is one of the major contributions of the Modern Synthesis that put together natural selection and Mendelian genetics under the umbrella of population genetics almost a century ago  and organized by the guest editors with the sponsorship of ountries , contribountries , the Ibeountries , encompaSolanum pimpinellifolium (Solanaceae) in South America. This allowed the authors to hypothesize about the origin of the species, which remained controversial. In addition, they identified the major ecological factors accounting for demographic behaviour and estimated the genetic offset, that is, whether the current genetic composition is maladaptive to future environments (Brasilianthus carajensis (Melastomataceae) from Amazonian Savannas. The authors not only characterized the population differentiation of a recently discovered species, but also employed genomic data to outline conservation/management actions to prevent the loss of unique genetic variation and maximize species resilience to future environmental change  in SE Spain. 4 grass Panicum hallii (Poaceae) across its distribution in North America (Mexico and USA) also merging RAD sequencing and species distribution models. The authors identified genetic clusters that followed the main ecoregions of the species across its range, and pinpointed South Texas as a major hotspot of genetic diversity with the co-occurrence of all genetic clusters and admixture between the two Panicum varieties of study.Furthermore, Lupinus angustifolius (Fabaceae) to two ecologically meaningful water availability treatments. The common garden experiment tested the effect of drought treatments on plant traits and allowed the authors to identify cogradient variation in drought-related traits, that is, plasticity to drought was in most cases in the same direction as among-population quantitative genetic differentiation (Geum triflorum (Rosaceae). Betula pendula (Betulaceae) in three common garden experiments across a substantial part of the species’ latitudinal range in Europe . In their study, P. hallii varieties  in a common garden experiment in South Texas (USA). In this case, the authors used an experimental approach as a complement to characterize the divergence between the two Panicum varieties of study, finding important differences between them in morphological features, cold stress tolerance and lifespan . On top of architectural and life-history data, gene expression data can also be used to assess genetic differentiation and local adaptation for samples from field populations, common garden experiments and/or reciprocal transplant experiments , understanding evolutionary processes still requires field studies of natural populations. In fact, differentiation is an all-encompassing process that eventually molds organisms’ phenotypic integration and their genetic make-up. Here, we gathered a collection of papers illustrating the relevance of traditional evolutionary studies focused on plant populations, but also highlighting the integration of classical biological disciplines and state-of-the-art molecular techniques into a unique toolkit for evolutionary plant research.A. thaliana; Based on the papers of this special issue, we envisage two promising future research lines to keep making progress in the discipline. First, it is time to bring together ecological genetics/genomics studies combining among- and within-population variation in genetic diversity and quantitative traits at large geographical scales (see We conclude by stressing the current serious challenges menacing the existence of plant biodiversity due to the unprecedented transformation of the planet at the beginning of the Anthropocene. The continuous differentiation process affecting the evolution of plants is probably experiencing dramatic alterations with unpredictable consequences. We need population differentiation studies, like the ones in this special issue, more than ever to comprehend the causes and consequences of rapid environmental changes on plant populations to foresee—and perhaps mitigate—their effects on plant biodiversity’s fate.


As part of a field study (NCT04473768) in children presenting with severe febrile illness to Kisantu hospital (DR Congo), we retrospectively compiled user experiences (not performance) with handheld diagnostic devices assisting triage: tympanic thermometer, pulse oximeter , hemoglobinometer and glucometer. Guidance documents for product selection were generic and scattered. Stock rupture, market withdrawal and unaffordable prices interfered with procurement. Challenges at implementation included environmental temperature, capillary blood sampling , calibration  and liability-oriented communication with a manufacturer. Instructions for use were readable and contained symbol keys; two devices had printed French-language instructions. Shortcomings were poor integration of figures with text and distinct procedures for the oximeter and its sensor. Usability interview revealed appreciations for quick results, visibility of the display and memory function (three devices) but also problems of capillary blood sample transfer, cleaning, too long of a time-to-results (respiratory rate) and size, fitting and disposal of thermometer probes. Pictorial error messages were preferred over alphanumeric error codes but interpretation of symbols was poor. Alarm sounds of the oximeter caused unrest in children and caretakers perceived the device as associated with poor prognosis. Severe febrile illness is a major global cause of under-five mortality and affects particularly children in sub-Saharan Africa ,2,3. SevTo facilitate recognition of danger signs, the World Health Organization (WHO) recommends the use of handheld diagnostic devices. These devices are used bedside and provide results within a few minutes. Examples are an oximeter to detect hypoxia or a glucometer to detect hypoglycemia. To improve rolling-out of diagnostic technologies, WHO developed recommendations that cover the complete life cycle of the handheld diagnostic devices, from development to disposal  7]..7].In this retrospective study, we compiled obstacles experienced throughout the lifecycle of handheld diagnostic devices used for hospital triage of children under five-years-old presenting with severe febrile illness in a low-resource setting. Experiences were reported from an end-user’s perspective with special attention for available guidance for selection of medical devices, instructions for use, usability and label comprehension.Plasmodium falciparum malaria, invasive Salmonella infections, anemia and malnutrition is high in children under five-years-old Procedure: SOP in Supplement S7Clinical implications: Rapid and bed-side measurement on capillary blood is important for identification of children who require urgent blood transfusion [Salmonella bloodstream infections [nsfusion  and for nsfusion . Severe fections ,37.For selection of devices and guidance in the procurement process, we consulted the WHO List of Essential Diagnostics , the WHOWe adhered to general criteria for point-of-care handheld devices in low-resource settings, i.e., they must be affordable, accurate, robust, rapid and user-friendly ,4,29. InAll devices were ordered and procured in Belgium and shipped in batch to Kisantu Hospital. All correspondence was done by the English-proficient principal investigator of the DeNTS study  supported by a research team competent in in vitro diagnostics.Instructions for use (IFU) were integrated into French language standard operating procedures (SOPs) by the principal investigator. Device-specific training of end-users at study initiation was based on these SOPs and organized by the principal investigator. Both IFU and SOPs were at the disposal of all end-users at the site of triage. After training, end-users were supervised by the principal investigator on-site during the first 6 weeks of the study. Hereafter, supervision was organized remotely by weekly teleconferences and ad hoc needs-based contacts , and as part of on-site supervisory visits. During the first 4 weeks of the study, a rotation system was used to ensure that all end-users got familiar with all devices. After these 4 weeks, each healthcare worker was assigned specific responsibilities to create one expert per study task. Nevertheless, all end-users continued to use all devices during weekend duties or moments of busy patient flow.We evaluated the accessibility  of IFU of the implemented handheld diagnostic devices. In addition, we assessed readability of the IFU instructions for (preparations of) the measurement with an online tool that determined the educational level needed to understand a French language text . FinallyAfter 10 months of use, we assessed the usability of the devices based on a semi-structured group interview  with eigWe used four handheld diagnostic devices , i.e., aWHO only provided detailed technical specifications for tympanic thermometers . For theThree devices were initially selected, but finally not ordered due to COVID-19 related stock rupture , high cost of ownership  and market withdrawal due to a programming error . Although this market withdrawal had preceded the order, the distributor accepted the order. The distributor (the only distributor of the device worldwide) required a minimum order of 20 pieces.Environmental temperature sometimes exceeded the maximum operating temperature (33 °C) of the tympanic thermometer causing non-function and a need to cool-down the thermometer. Overheating particularly occurred when the thermometer was transported in the pocket of a healthcare worker’s uniform, but also when transported in a separate, non-covered box. The multimodal oximeter failed to deliver results in-time with 6% (12/202) measurements lasting more than 5 min (see previous study ).Difficulties in capillary blood sampling and transfer of blood to strips and microcuvettes caused unreliable glucose and hemoglobin measurements at the start of the study. Poor repeatability of hemoglobin measurements (up to 2 g/dl differences) was linked with subtle underfilling of the microcuvette and “milking” of the finger during capillary blood sampling. Incorrect detection of hyperglycemia  was mostBlue dots in the hemoglobinometer’s microcuvettes were observed , which wFrench language IFUs in a printed booklet format were only available for the hemoglobinometer and glucometer . In the For the multimodal oximeter, IFUs from the oximeter and sensor were separate documents. As a consequence, no complete, step-by-step instructions for connecting, measuring and reading was available. In addition, figures on the use of the sensor were not integrated in the text. For the tympanic thermometer, the figures did not provide guidance on the alignment of the probe with the auditory canal, which is the most difficult step of the measurement.For the hemoglobinometer and the capillary glucometer, the IFU sampling procedure did not provide alternatives for cleaning the finger prick site when running water was not available on site, as was the case in our setting. The IFUs of both devices did not provide information about the optimal order to take samples from a single capillary finger prick in case of multiple testing  learned that the end-users considered maintenance and cleaning as feasible for all devices, although cleaning of electronic or measurement parts  was reported as delicate. End-users declared a preference for a rechargeable system instead of batteries for the glucometer and hemoglobinometer. In case of the thermometer, a screwdriver was needed to change the batteries. Quality control for the glucometer was declared complex and confusing, in contrast to the quality control for the hemoglobinometer which was considered as relatively easy.The label comprehension study revealed very limited comprehension of the meaning of the symbols labeled on the devices, their package and IFU. For 6, 6 and 3 out of 20 symbols, there were 0, 1 and 2 correct answers, respectively; median correct scores for nurses and physicians were 4/20, and 8/20, respectively .A detailed interview from all feedback and suggestions made during the usability interview is described All end-users, including the one who used it daily, reported that they struggled with the multimodal oximeter, particularly because of the occasionally long measurement times for the respiratory rate which disrupted patient flow at the triage. By contrast, they highly appreciated the short time-to-result of the thermometer, glucometer and hemoglobinometer. For these three devices, they also liked the memory function and large display of the results with someone clarifying that she “can even read the results without glasses”. End-users appreciated the self-explanatory error messages of the thermometer but struggled with the alphanumeric coded error messages of the glucometer and hemoglobinometer.End-users were in favor of the size and weight of the multimodal oximeter, glucometer and hemoglobinometer. They stated that the thermometer was relatively bulky and that they “had thought it would have additional functions given its size”. End-users further noted that the thermometer probes and oximeter sensor were not adjusted to the size of the smallest children. They also struggled with the single-use probes of the thermometer, because these were sometimes difficult to attach swiftly and impractical as they had to carry along a small waste bin to throw them away.Use of the multimodal oximeter to measure the oxygen saturation and heart rate was reported not difficult when children were calm. However, many children were agitated or got agitated during the measurement. Agitation complicated measurements by the multimodal oximeter, correct positioning of the thermometer and capillary blood sampling for the glucometer and hemoglobinometer. The interviewed staff further reported that the alarm sounds of the multimodal oximeter worsened agitation of the child and alerted the children’s caretakers who associated the oximeter with severe illness and poor prognosis. By contrast, some children got used to the thermometer during hospital follow-up and perceived it as a telephone toy.Overall, when asked if they would buy the device again irrespective of affordability, end-users reported that “they would pay a lot of money for the hemoglobinometer”. They would also buy the glucometer, but not the multimodal oximeter. They would buy the thermometer if there was a solution for the single use probes.In a field study addressing triage of children under five with severe febrile illness in a rural district hospital in sub-Saharan Africa, we used handheld diagnostic devices to guide detection of those in need of hospital admission and urgent antimicrobial and supportive treatment. Three devices were selected, but stock rupture, market withdrawal and unaffordable cost precluded their procurement. The four selected, procured and used devices  had challenges and shortcomings along their lifecycle, compromising fulfilment of the REASSURED-criteria .WHO guidance documents were generic (list of devices recommended per level of care) and not detailed. A few target product profiles compiled by other international actors and some independent device evaluations were retrieved, but information was scattered and fragmented. Target product profiles and technical specification sheets should be centralized and cross-referenced among international actors. In addition to guiding selection, independent product evaluations are valuable to understand performance and limitations. Although we did not retrieve independent product evaluations in the reference section of the IFUs assessed, they have certainly a place there .The transcutaneous hemoglobinometer was very attractive for triage given its non-invasiveness and potential speed, but we did not procure it because of a high fixed price (2 USD) per test, exceeding the acceptable  and desirable  price per test cited by the UNICEF hemoglobinometer target product profile for low-resource settings . The fiBased on present procurement experiences, we recommend that product-cycle oriented information is shared upfront and easily accessible online including a certificate of regulation, needs for calibration and maintenance and a list of common accessories and spare-parts. International shipments considerably increased total cost of ownership and complicated quality control (cold chain requirements of control fluid for hemoglobinometer) and calibration . Moreover, the fact that the distributor of ChARM respiratory rate counters was unaware of their withdrawal from market suggests poor communication with the manufacturer. National diagnostic strategies and governance have the potential to improve in-country supply ,64.The device implementation phase required high investments . This means that, when implementing such devices in a routine frontline diagnostic setting, prioritization and gradual adoption is key. The study staff prioritized the glucometer and hemoglobinometer, which provide actionable results and, in case of the glucometer, provides information that cannot be generated from the clinical exam.Not all IFUs were available in a printed French-language format and in the case of the tympanic thermometer and in particular the multimodal oximeter, figures and components could be better integrated and aligned with the written instructions. Readability of instructions for use was excellent, but was only assessed for the measurement instructions, which is the easiest and best structured section of instructions for use. The actual readability level of the procedure’s section of the IFU  represents the target for non-laboratory trained staff  but is aEnd-users preferred error messages depicted as graphical symbols rather than as alphanumeric codes and this apparently contrasted with the low scores in the label comprehension study. The low scores may partially be explained by the stand-alone versus contextual presentation of the symbols but comprehension of internationally recognized symbols is generally poor to moderate among non-trained participants ,66. SymbPivotal to usability and performance was the ease of specimen collection, which is particularly important during triage by healthcare workers with limited clinical expertise ,27. PresFailure of the thermometer in a too-hot environment confirmed the importance of robustness. Further, they appreciated the handy glucometer but disliked the bulky tympanic thermometer; they liked a large display of all devices the presence of a memory function and except the multimodal oximeter. Large font-sizes are particularly important in a low-resource setting, where impaired vision is often undercorrected ,72. The Of note, the usability interview revealed the importance of patients’ and caretakers’ experiences and perception. Children were afraid or became agitated by the long measurement and alarm sounds of the multimodal oximeter, which also caused unrest among caretakers, hampering also the other measurements. Moreover, caretakers perceived the multimodal oximeter as a sign of bad prognosis. These observations show the importance of assessing perceptions in the field-context of the intended use and user.A consistent point impacting affordability, delivery and access was the lack of an in-country distributor, which required logistics , impacted work  and entailed additional costs. In-country technical support services are highly recommended as they could also assist with on-the-job-training, appropriate use and integration in daily practice, calibration, quality control, maintenance and disposal .Correspondence with remote manufacturers occasionally caused misunderstanding. Upon a technical question (presence of blue dots in the hemoglobinometer’s microcuvettes), the manufacturer replied with a request for extensive and detailed technical and clinical information. Although sufficient detail is required for sound investigation of customers’ complaints and post-market surveillance , the levAs discussed above, quality control of the hemoglobinometer and calibration of the thermometer were challenged at product selection (clear upfront communication of requirements) and by issues of access and delivery . All devices except the multimodal oximeter had single-use consumables  requiring disposal and challenging environment-friendliness. End-users declared a preference for rechargeable integrated batteries  which facilitate stock management and decreases the inherent disadvantages if replaceable batteries .A major limitation of this study was its retrospective design. However, the study was embedded in a prospective clinical study (DeNTS study) commanded by the same principal investigator and study team. All incidents and associated decisions were documented as part of the DeNTS study, i.e., in preparatory communication during selection and procurement processes, SOPs, weekly reports, log sheets, case report forms etc. The perspective of the end-users might have been biased by the fact that they were part of a study team, which was in-depth trained and monitored. However, study nurses and physicians had similar qualification, background knowledge and experience as the other hospital staff. They were working in the same busy triage setting and focused on patient care rather than on device evaluation per se.As to generalizability, the present findings cannot as such be extrapolated to the health center level, where frontline healthcare workers are often less skilled and experienced ,74, whicThe assessment of usability, IFU and label comprehension in combination with reported field incidents allowed comprehensive evaluation. Little guidance is available on how to assess usability in low-resource settings but the semi-structured interview allowed detailed feedback and revealed multiple suggestions for device improvement. We did, however, not address sensitivity or specificity, which are key determinants of appropriate and effective devices.Screening and diagnosis are the bottleneck of appropriate patient care  and near


Variability is the source on which selective pressure acts, allowing genome evolution and adaptation. Large-scale comparative sequencing promises to reconstruct the evolutionary history of the human genome and to highlight the functional genetic differences between human and other species. Thanks to the recent advances in genome sequencing, assembling and studying complex regions of the genome enriched in segmental duplications and repetitive elements is not a limited process anymore. This is extremely important, on the one hand, in measuring the contribution of the emergence of novel genes in species diversification and, on the other hand, in evaluating the effect of evolutionary genomic rearrangements in human disease recurrence.BoPUB and NHX genes and their expression and adaptation in plants. In particular, Hu and colleagues performed genomewide identification and evolutionary analysis of the U-box protein family of Brassica oleracea (BoPUB genes) [BoPUB genes underwent a family expansion from the Arabidopsis thaliana to the B. oleracea genomes. RNA-seq transcriptome data of different pollination times revealed spatiotemporal expression specificity of the BoPUB genes. The study provides valuable information indicating that the BoPUB genes not only participate in the abiotic stress response but also are involved in pollination. Akram et al. instead performed a genomewide analysis of NHX genes in Gossypium barbadense in comparison with Gossypium hirsutum [NHX gene family, they can compare the gene regulation, expression pattern, and eventually their biological functions in cotton, providing valuable information explaining the molecular mechanism of Na+ transport for further functional studies of Gossypium NHX genes.This Special Issue consists of nine original research articles and two reviews covering various aspects of genome organization and gene family evolution. Two original research publications are focused on abiotic stress-related genes such as B genes) . The BoPhirsutum . The autT. pardalis Temminck & Schlegel and T. snyderi Abe, is largely controlled by an overlapping genomic segment (QTL). The QTL region contains no known genes responsible for the evolution of the scale-loss phenotype in other fishes. The genes used for the scale-loss phenotypes in the two Takifugu species are likely the same, but they are different from genes with the same function in distantly related fishes. The authors identified in the QTL region Fgfrl1 a gene predicted to function in a pathway known to regulate bone/scale development. Since Fgfr1a1, another member of the Fgf signaling pathway, has been implicated in the scale loss/scale shape in fish distantly related to Takifugu, the authors suggested that the convergence of the scale-loss phenotype may be constrained by signaling modules with conserved roles in scale development.Kim and colleagues’ study focused on the rapid evolutionary radiation of the pufferfish (Takifugu species) in marine ecosystems . The genAPOBEC3 locus.In recent years, technical advances in genome sequencing have greatly enhanced our ability to investigate the genetic differences within and between human populations and other species. Shen and colleagues developed QuicK-mer2, a method for studying copy number variation that enables the rapid construction of paralog-specific copy number maps from short-read sequence data . The autIn their review, Lallemand and colleagues described the evolutionary processes allowing the formation of duplicated genes and the various bioinformatic approaches that can be used to identify them in genome sequences . Indeed,POTE genes, a primate-specific gene family expressed in the prostate, ovary, and testis, as well as in several cancers, including breast, prostate, and lung cancers [POTE tandem duplication in marmoset that is misassembled and collapsed in the reference genome, thus revealing the presence of a second POTE copy. The study provides comprehensive insights into the evolutionary dynamics of the primate-specific POTE gene family, involving gene duplications, deletions, and long interspersed nuclear element (LINE) transpositions to explain the actual repertoire of these genes in human and primate genomes.Using single-molecule, real-time sequencing, in silico analyses, and molecular cytogenetics, Maggiolini and colleagues characterized the structure, copy number, and chromosomal distribution of the  cancers . This geStructural variants (SVs), including deletions and inversions, comprise a larger proportion of genetic differences between and within species, making them an important yet understudied source of trait divergence. Recent comparisons of short- and long-read sequencing technologies using benchmark human genomic datasets revealed that multiple genomes and combinatorial platforms are required for comprehensive SV discovery. Soto and colleagues performed long-range optical mapping and Oxford Nanopore Technologies long-read sequencing to characterize the structural variant landscape of two Pan troglodytes versus individuals . These nFGF4 gene in canids [Batcher and colleagues successfully used whole-genome sequence data to identify novel polymorphic retrocopies of the n canids . Gene reSaguinus oedipus, Callithrix argentata, and Alouatta belzebul, providing insight into the evolution of human chromosome 13. Furthermore, a review of previous cytogenomic literature data on chromosome 13 evolution in eutherian mammals showed a complex origin of the eutherian mammal ancestral karyotype, which has still not been completely clarified.Scardino and colleagues investigated the history of human chromosomes using comparative cytogenetic approaches . Human cInsights into the evolution of the modern eukaryotic cells come from the study of Diroma and colleagues . By applFinally, Antonacci and colleagues told us the tale of the T cell receptor (TR) loci in the adaptive immune response . T lymphOverall, the papers in this Special Issue cover various aspects of genome organization and gene family evolution and show the importance of combining the use of different methodological approaches to provide an overview of the dynamic and plasticity of genome architecture in both the animal and plant kingdoms.


As a common parasitic disease in animals, coccidiosis substantially affects the health of the host, even in the absence of clinical symptoms and intestinal tract colonization. Gut microbiota is an important part of organisms and is closely related to the parasite and host. Parasitic infections often have adverse effects on the host, and their pathogenic effects are related to the parasite species, parasitic site and host-parasite interactions. Coccidia-microbiota-host interactions represent a complex network in which changes in one link may affect the other two factors. Furthermore, coccidia-microbiota interactions are not well understood and require further research. Here, we discuss the mechanisms by which coccidia interact directly or indirectly with the gut microbiota and the effects on the host. Understanding the mechanisms underlying coccidia-microbiota-host interactions is important to identify new probiotic strategies for the prevention and control of coccidiosis. Eimeria ,antimicrobial peptides (AMPs), regenerating islet-derived protein 3, lysozymes, and other factors . Eimeriad MUC5ac , resultiecretion . The nume stages . Microore stages , use mucridiales , whose gn of MUC . The typn of MUC , and botn of MUC . TherefoEimeria praecox, several genes were downregulated, including those that encode antimicrobial peptide 2 and the cationic, anionic, and L-type amino acid transporters  are elicited following infection with parasites . Howeverytokines , therebyEimeria ninakohlyakimovae infected goats and revealed the increased levels of specific IgG, IgM, and IgA during the host immune response. By measuring the content of immunoglobulins and gut microbiota in inflammatory bowel disease patients, it was observed that IgG, IgM, and IgA had a positive correlation with Enterobacteriaceae and Enterococcus; while a negative correlation with Lactobacillus and Bifidobacterium. This indicates that IgM, IgG, and IgA are closely related to the imbalance in the gut microbiota, which may be caused by changes in the proportion and quantity of gut microbiota, leading to disruption of the intestinal mucosal microecological balance and abnormal immune responses. However, humoral immune reactions cannot eliminate primary coccidial infections  known to participate in the regulation of complex networks, thereby activating the migration of immune cells to the site of infection , which are the most widely and intensively studied end product of intestinal metabolism, play an important role in metabolism . These md others . Howeverd others . Acetic ntestine . A reduche cecum . In addihe cecum . SCFAs ahe cecum , which iEimeria necatrix, Eimeria mitis, and E. maxima  than in the later sexual stages. In coccidia-infected animals, the amount of sIgA is increased, which prevents microorganisms from residing and multiplying in the mucosal epithelium. sIgA can inhibit the invasion of bacteria in epithelial cells, increase the diversity of gut microbiota, and promote immune responses in intestinal epithelial cells . IL-22 de extent . Reg IIIbacteria . The immLactobacillus and Megasphaera. In growing pigs, a 10% reduction in the protein level decreased the diversity of ileal and colonic flora  of Lactobacillus rhamnosus inhibited the sporulation of oocysts, which demonstrated the anti-coccidial activity of SCS , which provided protection against Eimeria infection in broilers  How do probiotics regulate the gut microbiota to resist coccidia? (2) How do probiotics act on the intestinal biological barrier to exert anticoccidial effects? (3) What are the active ingredients of probiotics against coccidia? (4) Which probiotic has the best anticoccidial effect? In summary, a comprehensive understanding of the molecular mechanisms by which probiotics exert their beneficial effects on the host against coccidial infection is required for the development of highly effective probiotic formulations that can replace antibiotics for the prevention and control of coccidiosis.In recent years, with the development of high-throughput sequencing technology, research on the interrelationship between the gut microbiota and diseases has progressed, and an increasing number of researchers have recognized the important role of gut microbiota in disease onset, progression, treatment, and prognosis. Although the mechanisms by which coccidia and intestinal microbiota interact are not well understood, this review analyzed the different aspects of their interactions. Coccidia share the intestinal environment with microbiota and directly antagonize commensal bacteria. In addition, coccidia indirectly affect the intestinal microbiota. Mechanical mucosal damage , chemical mucosal damage  and disruption of the immune system provide conditions suitable for the growth of conditionally pathogenic bacteria, leading to changes in the intestinal microbiota. The addition of probiotics directly or indirectly impair coccidia development by improving the intestinal microbiota.Coccidia-microbiota-host interactions form a network of mutual constraints. For example, coccidial infection causes an imbalance in the intestinal microbiota, which not only leads to a decrease in food intake and impaired absorption but also increases the susceptibility of the organism to secondary infections. Conversely, the disruption of host health increases the number of pathogenic bacteria and impairs the intestinal mucosal barrier function and the ability of the immune system to target coccidia, resulting in a more serious coccidial infection. Coccidia, the microbiota and the host simultaneously interact, and a change in one factor may affect the entire network.In conclusion, a holistic approach is needed to gain a better understanding of the mechanisms underlying coccidia development and infection. However, most studies on coccidiosis have focused on avian species, with limited studies on ruminants. With the development of intestinal microbiota sequencing technology in recent years, we can improve our understanding of the mechanisms contributing to coccidia-microbiota-host interactions and provide a theoretical basis for the control of coccidiosis.CL wrote the manuscript. CN and YY revised the manuscript. All authors read and approved the final version of the manuscript for publication.This work was supported by the Earmarked Fund for China Modern Agro-industry Technology Research System (No. nycytx-38) and National Key R&D Program of China (No. 2018YFD0502100).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


MiRNA is a type of small non-coding RNA, by regulating downstream gene expression that affects the progression of multiple diseases, especially cancer. MiRNA can participate in the biological processes of tumor, including proliferation, invasion and escape, and exhibit tumor enhancement or inhibition. The tumor immune microenvironment contains numerous immune cells. These cells include lymphocytes with tumor suppressor effects such as CD8+ T cells and natural killer cells, as well as some tumor-promoting cells with immunosuppressive functions, such as regulatory T cells and myeloid-derived suppressor cells. MiRNA can affect the tumor immune microenvironment by regulating the function of immune cells, which in turn modulates the progression of tumor cells. Investigating the role of miRNA in regulating the tumor immune microenvironment will help elucidate the specific mechanisms of interaction between immune cells and tumor cells, and may facilitate the use of miRNA as a predictor of immune disorders in tumor progression. This review summarizes the multifarious roles of miRNA in tumor progression through regulation of the tumor immune microenvironment, and provides guidance for the development of miRNA drugs to treat tumors and for the use of miRNA as an auxiliary means in tumor immunotherapy. TME contains a variety of cells, including fibroblasts, endothelial cells, mast cells, and immune cells (The tumor microenvironment (TME) facilitates growth of tumor cells ne cells . Fibroblne cells , while ene cells . Immune ne cells . Among tne cells . DC funcne cells . In addine cells . Many stne cells . It is tMicroRNA (miRNA), as a non-coding small RNA, is produced from precursor RNA and is usually composed of 19-25 nucleotides that can target and regulate downstream gene expression . AbnormaMiRNAs are involved in most aspects of cancer biology, such as apoptosis, proliferation, angiogenesis, invasion/metastasis, and tumor stemness. Some miRNAs display tumor suppressor effects, such as miR-29b, which inhibited the expression of antiapoptotic genes Mcl-1 and Bcl-2, and promoted spontaneous apoptosis of tumor cells . Other mvia exosomes to immune cells, thus exosomes as an important tool for miRNA transport can also affect the immune microenvironment. The diverse roles of miRNA on different tumor cells will be discussed in the following sections.In the TME, there is a close relationship between tumor cells and their surrounding immune cells, and miRNA is thought to connect these cell types. Abnormal expression of miRNA in tumor cells can affect the immune microenvironment. For example, miRNA can control tumor cells to produce chemokines or cytokines, which then affect the aggregation and expansion of immune cells. Contrarily, tumor cell miRNA can be transported TAM are known to participate in tumor cell growth, survival, invasion, metastasis, angiogenesis, inflammation, and immunoregulation. Moreover, studies have found that miRNA are also involved in the process of regulating TAM to affect tumors.MiRNA can influence phenotypic changes and the polarization of macrophages Figure 1via transfection with miR-125a/b reduced the stemness of liver cancer cells by targeting CD90 and inhibited the proliferation of these cancer cells  and activate the PI3K/Akt pathway. This enhanced the immunosuppressive activity of MDSC, and the enhanced expression of metalloproteinases (MMPs) promoted the escape and metastasis of tumor cells . MoreoveIn summary, these research findings show that tumor cells or MDSC abnormally expressing miRNA can accelerate the formation of MDSC in the TME and enhance the immunosuppressive activity of MDSC. Thus, miRNA has a critical role in the function and development of MDSC in the TME Table 3.In the previous chapters, we have summarized the specific functions and characteristics of Treg in the TME. Studies have found that miRNA are engaged in the process of regulating Treg in the TME, reflected mainly through miRNA affecting the differentiation, function, and aggregation of Treg in the TME. In addition, other studies have shown that miRNA also play an important role in the regulation and balance of Treg and their effect on tumor growth.in vitro experiments demonstrated that miR-21 could inhibit the proliferation of CCR6+ Tregs, mainly through targeting PENT and then activating the AKT pathway , National Key Research and Development Project (No. 2017YFD0400303).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Prostatitis is a common disease in adult males, with characteristics of a poor treatment response and easy recurrence, which seriously affects the patient’s quality of life. The prostate is located deep in the pelvic cavity, and thus a traditional infusion or other treatment methods are unable to easily act directly on the prostate, leading to poor therapeutic effects. Therefore, the development of new diagnostic and treatment strategies has become a research hotspot in the field of prostatitis treatment. In recent years, nanomaterials have been widely used in the diagnosis and treatment of various infectious diseases. Nanotechnology is a promising tool for 1) the accurate diagnosis of diseases; 2) improving the targeting of drug delivery systems; 3) intelligent, controlled drug release; and 4) multimode collaborative treatment, which is expected to be applied in the diagnosis and treatment of prostatitis. Nanotechnology is attracting attention in the diagnosis, prevention and treatment of prostatitis. However, as a new research area, systematic reviews on the application of nanomaterials in the diagnosis and treatment of prostatitis are still lacking. In this mini-review, we will highlight the treatment approaches for and challenges associated with prostatitis and describe the advantages of functional nanoparticles in improving treatment effectiveness and overcoming side effects. Prostatitis is one of the most common urogenital diseases and mainly manifests as hypogastrium, perineum, scrotum, urethra and penis pain, and even bladder irritation, seriously affecting the patient’s quality of life . AccordiProstatitis is mainly divided into 2 class I acute bacterial prostatitis, class II chronic bacterial prostatitis, class III chronic prostatitis/chronic pelvic pain syndrome, and class IV asymptomatic inflammatory prostatitis . In addiThe pathogenesis of prostate disease is complex with numerous influencing psychological factors, including pathogen infection, sex hormone imbalance, urination dysfunction, inflammation, and abnormal immune response , among wEscherichia coli , computed tomography (CT), and ultrasound used today after a long period of exploration and growth. Although these imaging techniques have different imaging principles, they all observe tiny lesions in a noninvasive manner, providing excellent images of humans due to their unique advantages. However, they have some inherent limitations. For example, magnetic resonance imaging has an insufficient spatial resolution, leading to low sensitivity [46]. Therefore, many contrast agents have been developed to improve the contrast between normal tissue and prostate lesions and thus improve the diagnostic accuracy.The cause of prostatitis is multifactorial, and the disease course is long. The most commonly used mode in clinical practice, rectal administration, may cause damage to intestinal mucosa due to unstable drug absorption. Therefore, the treatment of prostatitis requires good imaging performance, strong compatibility, a high universality of imaging technology, and a high bioutilization of pharmaceutical preparations. Advances in nanotechnology have facilitated the development delivery systems to overcome prostate-related disorders. Advantages of nanocarrier preparations include the combination of a variety of drugs, including biomacromolecule drugs; reduced degradation of unstable drugs and slow and controlled release; and increased residence time of relevant drugs to avoid frequent injections to meet the needs of prostatitis treatment . In addiCampylobacter infection is one of the most common foodborne infections in humans, and broilers are the main source of Campylobacter infection  is a receptor expressed on small intestinal cells and antigen-presenting cells (APCs). The combination of APN-specific targeting drugs with vaccine antigens significantly stimulates the immune response in the intestinal mucosa. Bakshi [53] constructed an anti-porcine APN nanoantibody with the Fc domain of conventional antibodies to form a bivalent fusion protein that triggered the intestinal IgA response after oral administration and confirmed the potential of vaccine antigen carriers . Modern Anti-inflammatory and antimicrobial agents are the two main strategies for the treatment of urethral inflammation. We will summarize the application of nanomaterials in the treatment of prostatitis from antibacterial and anti-inflammatory aspects.Pathogenic microorganisms such as viruses, fungi and bacteria cause prostatitis, among which bacterial infection is the main pathogenic factor causing prostatitis. The number of antimicrobials used to eradicate type II chronic bacterial prostatitis is very limited. Treatment of CBP is hampered and challenging because most antimicrobial agents have a poor ability to penetrate infected prostate fluids and tissue . AnotherProteus novelis and Proteus vulgaris fliL genes, which are clinically useful for urinary tract infections, thus exerting an anti-infection effect (Bacterial biofilms are an important barrier that promote bacterial self-protection and an important mechanism of therapeutic tolerance. Nanomaterials have shown unprecedented advantages in destroying bacterial biofilms. Li et al. realized the antimicrobial effect of the biofilm microenvironment response by designing antibiotic quantum dots . In uretn effect . As in-dA large number of experiments have proven that the pathogenesis of prostatitis is closely related to the inflammatory microenvironment . However2 nanoballs promoted prostate urethral epithelial changes, enhanced the antioxidant capacity by inducing Ikk expression in macrophages, where I kappa B predominates, and p65 phosphorylation to inhibit oxidative stress and induce macrophages to differentiate into M2 phenotypes, reducing inflammatory reactions (In addition, oxidative stress and inflammation are closely related to the immune responses that maintain homeostasis in the body. Oxidative stress is not only an important feature of inflammation but also a cause of inflammation , Liu et. Seleniueactions . Nanoseleactions .2, Fe3O4 and nano silver hydrosol, have been experimentally proven to be useful in the prevention, diagnosis and combined treatment of prostatitis. Although nanomaterials have achieved impressive results in experimental studies, their clinical conversion still faces significant obstacles. First of all, the metabolic pathway of some inorganic nanomaterials in vivo is not clear, and the cumulative toxicity is high. For example, silver nanoparticles, commonly used in urinary tract infections, accumulate in the body and cause liver and kidney toxicity. Secondly, traditional nanomaterials, as exogenous substances, are easy to trigger the immune response of the body, and are easily cleared by the immune system. In addition, nanomaterials as contrast agents also have the defects of low resolution and limited imaging depth. These problems greatly limit the application of nanomaterials in the clinical diagnosis and treatment of prostatitis. How to overcome the above obstacles has become the current research focus of nanomedicine.The causes of prostatitis are complex and include pathogen infections, inflammation, free radicals, an abnormal immune response, sex hormone imbalance and so on. The treatment and diagnosis of prostatitis is facing great challenges. Nanomaterials with anti-inflammatory effects, such as CeOThe latest research progress summarized in this review, and it is not hard to find out the future research direction in this field. First, nanotechnology will promote the development of clinical diagnosis of prostatitis, especially molecular imaging research based on multimodal imaging technology will further improve the sensitivity and specificity of diagnosis. At the same time, nanotechnology will also facilitate the development of liquid biopsies, which are called upon to combine body fluid detection with medical imaging. Secondly, nanomedicine will break away from the traditional nanomaterials to the clinical application, which mainly depends on the development of new organic or biological sources of nanomedicines. The emergence of natural nanocarriers, such as exosomes, eliminates the immunogenicity and metabolic risks of traditional nanomaterials, making their clinical applications possible. Third, nanotechnology combined with machine learning can help achieve multifunctional integration and personalized diagnosis and treatment. Nanomedicine will undoubtedly revolutionize prostatitis diagnosis and treatment.