As SARS-CoV-2 infections and death counts continue to rise, it remains unclear why some individuals recover from infection, whereas others rapidly progress and die. Although the immunological mechanisms that underlie different clinical trajectories remain poorly defined, pathogen-specific antibodies often point to immunological mechanisms of protection. Here, we profiled SARS-CoV-2-specific humoral responses in a cohort of 22 hospitalized individuals. Despite inter-individual heterogeneity, distinct antibody signatures resolved individuals with different outcomes. Although no differences in SARS-CoV-2-specific IgG levels were observed, spike-specific humoral responses were enriched among convalescent individuals, whereas functional antibody responses to the nucleocapsid were elevated in deceased individuals. Furthermore, this enriched immunodominant spike-specific antibody profile in convalescents was confirmed in a larger validation cohort. These results demonstrate that early antigen-specific and qualitative features of SARS-CoV-2-specific antibodies point to differences in disease trajectory, highlighting the potential importance of functional antigen-specific humoral immunity to guide patient care and vaccine development.  •Limited early differences were observed in titers and neutralization across groups•Five antibody features could collectively differentiate convalescents and deceased•A shift in the balance of spike versus nucleocapsid immunity separated the groups•Spike-specific phagocytic and complement fixing activity was enriched in convalescents Although most SARS-CoV-2-infected individuals experience mild disease, a significant fraction of individuals become severely infected. Early biomarkers that predict outcome are urgently needed. Atyeo et al. demonstrate that distinct acute SARS-CoV-2 humoral immune responses exist across severely ill individuals who ultimately convalesce or pass away. SARS-CoV-2 is the newest coronavirus to cross into the human population . MillionAlthough antibody responses and T cells have been linked to disease resolution , and neuAntibody dynamics during the acute window of infection have been linked to differential outcomes across infections, including HIV , influenAcross infectious diseases, pathogen-specific antibodies can serve as biomarkers of infection and aid with early control and clearance of infection by blocking host-pathogen interactions and/or recruiting innate immune functions . To inveTo profile the SARS-CoV-2-specific humoral immune response, we performed systems serology to determine the biophysical and functional characteristics of SARS-CoV-2-specific antibodies that recognize the SARS-CoV-2 S, the S-derived receptor-binding domain (RBD), and N. Titers of SARS-CoV-2-specific isotypes and subclasses, Fcγ-receptor binding profiles, neutralization, as well as antigen-specific innate effector functions were measured. Heterogeneous responses were observed across both populations , and conBeyond univariate differences, emerging data point to a critical role of humoral immune response coordination as a predictor of protection in some infections . Given tConversely, convalescents overall displayed a more uniform correlation profile across subclass and isotype responses and antibody effector function. However, although neutralizing antibody responses were co-induced with isotype and subclass and effector functions among individuals who died, neutralizing antibody responses were largely inversely correlated with all antibody responses among individuals who recovered, suggesting a divergent evolution of the antigen-binding and constant domain of the antibody across these populations. These data highlight multiple early functional differences in SARS-CoV-2-specific humoral immunity between the groups.To further probe the overall humoral profile between groups, the mean percentile of each antibody metric was determined across SARS-CoV-2 antigen specificities for both populations B. NightiGiven the unique correlation and immunodominance profiles across the groups A and 2B,Given that the feature down-selection algorithm selects a minimal set of features to avoid overfitting, a co-correlates network was used to explore additional features that may distinguish these two groups F. A largCollectively, the data point to a shift in immunodominance of S versus N functional antibody responses. To test this hypothesis, we next compared the overall ratio of S:N-specific antibody isotypes, subclasses, and functions across the groups A. As expCellular and humoral immune responses have been linked to protection against several coronaviruses . ImportaIn SARS-CoV-1 and SARS-CoV-2 infection, N is highly immunogenic, with N-specific humoral immune responses arising concurrently with S-specific humoral immunity . HoweverIt is well known that the timing of sampling may influence humoral profiles, where sampling time could result in comparison of immature versus mature immune responses. Despite the sampling differences in the group, comparable titers were observed across convalescents and individuals who ultimately passed away. Moreover, similar overall functional profiles were also observed, suggesting that the humoral immune responses were comparable in magnitude across the two groups. Additional analysis of the influence of sampling time on the spread of the antibody profiles in the PLSDA highlighted a minimal influence of time from symptoms on overall antibody profile variation, and the time of sampling exhibited a minimal predictive power in classifying individuals into convalescents or deceased. However, longitudinal analyses will be illuminating, providing further information regarding the evolving humoral immune response that tracks with protection from infection.Emerging data point to higher mortality among the elderly and across genders . Along tAlthough S-specific antibodies able to recruit NK cell activity were expanded in individuals who passed away, pointing to a potentially negative influence of NK cells, coordination of NK cell and phagocytic activity was enriched among convalescents. These seemingly contradictory data point to the potential importance of synergy between innate immune effector functions. Although NK cells have been implicated in protection  and pathThere are a number of limitations in this study. First, because these samples were collected early during the COVID-19 pandemic in the United States, the Seattle study included a small number of participants, and the groups were not age- or sex-matched. Confounding factors such as timing of sampling, sex, and age are known to influence SARS-CoV-2 infection and disease trajectory. Although antibody profiles clearly segregated individuals who survived compared with those who did not survive, more limited variation in antibody profiles was observed across age, sex, viral load, and days from symptom onset. However, among the co-morbidities, age was the second major driver of variation in antibody profiles, pointing a potentially critical role of age-associated defects in Fc variation that may contribute to altered antiviral immunity to SARS-CoV-2 and beyond. The larger validation cohort from Boston identified a similar humoral signature that discriminated survivors from non-survivors, highlighting the conserved nature of this immunological signature independent of demographic characteristics. Although this study only attempted to understand the humoral disparities between convalescent and deceased individuals in a cohort of severely infected individuals, future studies may attempt to define humoral profiles able to further classify individuals across the clinical trajectory spectrum ranging from asymptomatic to severe disease.Collectively, the data presented here argue for the evolution of distinct antigen-specific and functional humoral immune responses early in SARS-CoV-2 disease. Although further analysis of longitudinal cohorts may provide more mechanistic insights into the specific role of antibodies in control and clearance of infection, here we validated an early functional humoral immune signature that appears to predict disease progression across two distinct cohorts. Linked to emerging animal model experiments, the correlates defined here may provide key mechanistic insights to guide therapeutic and vaccine design efforts.galter@partners.org).Further information and requests for resources and reagents should be directed to and will be fulfilled by the Lead Contact, Galit Alter  replicates and the average value was used throughout the study. This study was approved by the University of Washington Human Subjects Division Institutional Review Board.2.Primary immune cells were isolated from fresh peripheral blood from healthy human volunteers collected by the MGH Blood bank or the Ragon institute. The study was approved by the MGH Institutional Review Board. All subjects were over 18 years of age and provided informed consent. All samples were completely de-identified prior to use. Human NK cells and neutrophils isolated from fresh peripheral blood were cultured in RPMI supplemented with with 10% fetal bovine serum, L-glutamine, penicillin/streptomycin and maintained at 37°C, 5% CO2 in RPMI supplemented with 10% fetal bovine serum, L-glutamine, penicillin/streptomycin and 0.01% b-mercaptoethanol.THP-1 cells (ATCC) were grown at 37°C, 5% COAntigen-specific antibody subclass, isotype, sialic acid, galactose and Fcγ-receptor (FcγR) binding levels were assessed using a 384-well based customized multiplexed Luminex assay, as previously described  RelativeFor the functional analysis of plasma samples, bead-based assays were used to quantify antibody-dependent cellular phagocytosis (ADCP), antibody-dependent neutrophil phagocytosis (ADNP) and antibody-dependent complement deposition (ADCD), as previously described ; Data S1For detection of antibody-dependent NK cell activity, an ELISA-based approach was used, as described . BrieflyThe 2019-nCoV pseudoviruses expressing a luciferase reporter gene were generated as described previously  Yang et. BrieflyAll analyses were performed using python version 3.6.8 with statistical and machine learning packages . NetworkThe classification models were trained to distinguish convalescent and deceased groups with a minimal set of features, to avoid overfitting. PBS controls was subtracted from all features, Fc array features were log transformed, and all data was scaled and centered. Antibody features including sex and interventions  were incThe models were built using a backward feature elimination for selection and then classified using the minimal set of features which maximize accuracy . Models Performance and robustness of the model was contrasted with negative control models constructed from permuted data and randomly selected size-matched features, with multiple iterations of fivefold cross-validation used to generate classification accuracies. These control models were generated 100 times. The permuted control was generated in the same process as above shuffling labels randomly for each repetition. Size-matched features were chosen at random for each cross-validation step within each repetition. Predicted and true outcomes were compared to determine accuracy. Robustness was defined as the effect size of the distributions (Cliff’s Δ), and the exact P values of the tail probabilities of the true distributions within the control distributions. Reported are the median p values across twenty independent cross-validation replicates .Correlation networks were constructed to visualize the additional humoral immune features that were significantly linked to the selected minimal biomarkers, to provide enhanced insights into the biological mechanisms by which antibodies may provide protection following infection. In brief, antibody features that were significantly correlated with a Holms-Bonferroni correction to the final selected PLS model selected-features were defined as co-correlates. Significant spearman correlations above a threshold of |r| > 0.5 were visualized within the networks.Using the selected features from the original model a new PLSDA model was trained excluding a single outlier at a time in a fivefold cross validation framework. This process was repeated three times, each time generating a unique ROC curve as the top 3 individual outliers were removed. Using these cross validated ROC curves the mean performance and variation were assessed and are summarized as area under curve.In order to evaluate S versus N ratios, first ratios for each feature were defined separately by simply dividing S-responses over N-responses for every given feature. S:N ratios were visualized by log2 transformation for ease of interpretation. Differences across convalescents and deceased were then tested with a one-sided Mann-Whitney U test and a Holm-Bonferroni multiple hypothesis correction criterion.In order to address whether the overall S-response was enriched over N-responses in the convalescents across all features tested, all data was background corrected and z-score normalized. Then the number of S-features which were greater than their N-counterparts across every feature were summed. This analysis yielded a distribution of individual S greater than N scores for each group and statistical differences were assessed using a one-sided Mann-Whitney U test.


Dear editor,Many recent COVID-19 series have reported arterial or venous thrombosis  . Here, wOn April 3, 2020, a 63-year-old female presented to the emergency department because of aphasia and right hemiplegia. She had a 12-day history of fever, cough, and anosmia. Her husband was hospitalized in intensive care for confirmed COVID-19 acute respiratory distress syndrome (ARDS). Brain MRI showed a large left temporal brain hemorrhage and a suspicion of CVT confirmed on a venous brain CT scan and chest CT showed typical COVID-19 patchy ground-glass opacities in both lungs  and high ferritin levels (1427 μg/L).The nasopharyngeal and bronchial samples were negative for SARS-CoV-2.Most common causes of genetic thrombotic disorders and antiphospholipid antibody syndrome were excluded.The patient was started on an intravenous curative dose of heparin anticoagulation.Electroencephalograpy (EEG) showed background theta activity unreactive to nociceptive stimulus, with pseudo-periodic activity of a short period composed of slow di-phasic waves irradiating towards the anterior regions Fig. . AlthougOn April 17th, brain CT scan revealed a new intracranial contralateral bleeding most likely following contralateral venous thrombosis despite being properly treated with intravenous heparin. Venous angiography showed persistent left thrombosis (Fig. On April 29th, the patient died following therapeutic limitation after ethical consultation group expertise.Although both samples were negative for SARS-CoV-2, we considered the patient infected by it, given the initial symptomatology, the confirmed infection in one relative, the specific aspect of the thoracic CT scan , and theIn addition to the left lesion temporal focus observed on the EEG, the background activity and paroxysmal activity describes atypical patterns, which can be mistaken with persistent epileptic activity. However, we believe it is compatible with newly described patterns of specific COVID-19 encephalopathy .Overall, this case suggests that practitioners should be aware of the possibility of a CVT in this novel COVID-19 context, especially during the post-viral period.


In TCGA, 50 miRs showed significant association with survival. Among the top ranked prognostic miRs were members of the two miR families miR-221/222 and miR-17-92. Loss of miR-221/222 was correlated with improved prognosis in both cohorts  and was an independent prognostic marker in a multivariate analysis considering demographic characteristics ), molecular markers (O-6-methylguanine-DNA methyltransferase (MGMT) methylation, IDH mutation status) and PORT as co-variables. The prognostic value of miR-17-92 family members was ambiguous and in part contradictory by direct comparison of the two cohorts, thus warranting further validation in larger prospective trials.MicroRNAs (miRs) are non-coding master regulators of transcriptome that could act as tumor suppressors (TSs) or oncogenes (oncomiRs). We aimed to systematically investigate the relevance of miRs as prognostic biomarkers in primary glioblastoma multiforme (GBM) treated with postoperative radio(chemo)therapy (PORT). For hypothesis generation, tumor miR expression by Agilent 8x15K human microRNA microarrays and survival data from 482 GBM patients of The Cancer Genome Atlas (TCGA cohort) were analyzed using Cox-PH models. Expression of candidate miRs with prognostic relevance  was validated by qRT-PCR using Taqman technology on an independent validation cohort of GBM patients ( Glioblastoma multiforme (GBM) is the most frequent and aggressive form of primary brain tumors in adults, with an annual incidence of 3.2 per 100,000 . The staMicroRNAs (miRs) are small non-coding RNA molecules that consist of about 22 nucleotides that are able to repress gene expression by posttranscriptional decay of target mRNA. The miR molecule is incorporated into the RNA-induced silencing complex (RISC), which then targets the 3’-untranslated region of its target mRNA. Depending on the grade of complementarity between the miR seed sequence and the mRNA, it is either degraded or its translation is repressed. Missing the necessity for perfect complementarity between the miR and its target mRNA allows for the regulation of one target gene through several miRs as well as the regulation of several genes by one single miR ,24,25,26The Cancer Genome Atlas’ (TCGA) primary GBM cohort currently presents the largest repository of integrative molecular and clinical data for GBM and is frequently used for the hypothesis generation and validation of newly discovered biomarker . HoweverPatient characteristics are shown in In both the TCGA and HD cohort, clinical covariates age and KPI as well as treatment with TMZ and PORT showed significant association with OS . IDH1 muTreatment and molecular covariates showed expected behavior with regard to outcome. Missing data, especially in the TCGA cohort, however, increases uncertainty in the interpretation of the findings.p-value < 0.05) could be detected  are shown in We evaluated miR expression in the TCGA-GBM cohort (above upper vs. below lower quartile expression defined groups) with regard to their prognostic value. For a total of 50 miRs, a significant survival difference . In line with the training cohort, patients of the HD cohort with tumors expressing high levels of miR-221 or miR-222 showed a significantly decreased OS compared to patients expressing low levels of miR-221 or miR-222 C,D. MediFor all investigated members of the miR-17-92 cluster , a significant prognostic separation could be observed showing unfavorable prognosis in the high-expressing group . This coComparison of patients’ characteristics of the TCGA and the HD cohort revealed a certain degree of cohort heterogeneity, as outlined in For members of the miR-17-92 cluster, the subgroup analyses in TCGA and HD yielded more heterogeneous results. Significant results were achieved in only 4 to 8 out of 18 (TCGA) and 3 to 5 out of 16 (HD) subgroups . HoweverThus, covariates putatively modulating the effect of miRs could not be identified in either multivariate analyses or in these subcohort analyses. n = 482) and HD validation cohort (n = 109). MiR-221 and miR-222 are clustered together on chromosome Xp11.3 and have similar target specificity [We report the expression of miR-221 and miR-222 as important prognostic factors in both the TCGA training  ,63,64. COur survival analyses of members of the miR-17-92 cluster yielded contradicting results: within the TCGA cohort, high expression was associated with longer OS in univariate and multivariate analyses. In contrast, the HD cohort showed shorter survival for patients with tumors expressing high levels of miR-17-92 members. In Studies exploring the effect of miRs tend to be more heterogenous and may even produce contradictory results, which might arise from a number of issues, including the following: small patient cohorts with high uncertainty of clinical data from pathological annotation to treatment homogeneity; technique immanent problems in microarray-based screens , which are further complicated by the short sequence with high sequence homology across a miR family and most importantly the often low abundance of miRs, leading to high CT values in PCR-based methods and close to background microarray intensities. These factors might limit the reproducibility of hypotheses generated in the TCGA cohort, as found for the miR-17-92 in our study. Therefore, reliable analyses require uniformly treated patients, valid methods for verification of screening results, and patient groups of sufficient size. Additionally, further mechanistic studies investigating the cellular functions of miR-221/222 and miR-17-92 in GBM are needed. Interestingly, both of the investigated miR families are reported to increase GBM cell invasiveness, while infiltrative growth is characteristic for GBM cells and was postulated to be linked to enhanced radio-resistance of GBM cells . While hn = 482) and the HD (n = 109) cohort. Ambiguous results regarding the expression of members of the miR-17-92 family require further exploration. Taken together, miR-221/222 are promising prognosticators of OS in primary GBM, and could be confirmed as prognostic markers in two independent cohorts, namely the TCGA .Tumor samples were acquired through biopsy/surgical resection. Samples were assessed by board-certified neuropathologists for diagnosis of GBM and tumor fraction of at least 60%. IDH1 mutation and MGMT promotor methylation status were determined as previously described ,78,79. ® RNA Isolation Reagent  or AllPrep® DNA/RNA/Protein Mini Kits  in combination with RNeasy® MinElute® Cleanup Kit . RNA concentrations were measured with the NanoDrop 1000 Spectrometer , and RNA quality control was performed with the 2100 Bioanalyzer system . All steps were performed according to the manufacturers’ protocols. Total RNA from TRIzol®-processed samples and supernatant from AllPrep® purified samples were used for further analyses. RNA extraction from surgically acquired specimens was performed using TRIzol® MicroRNA Reverse Transcription Kit  was used, and for quantitative real-time PCR TaqMan® Universal PCR Master Mix II (no UNG)  was used. Primers were ordered at TaqMan® (Thermo Fisher Scientific): hsa-miR-221 (#000524), hsa-miR-222 (#002276), hsa-miR-17-5p (#00393), hsa-miR-18a (#002422), hsa-miR-19b (#000396), controls RNU44 (#001094), and U6_snRNA (#001973). Measured CT values were normalized against the arithmetic mean of controls (RNU44 and U6_snRNA). Data were further virtual pool normalized to the arithmetic mean of each miR. Expression of miRs was quantified by real-time PCR on an ABI 7900 system (Life Technologies). For reverse transcription, TaqManStatistical analyses were conducted in R v. 3.6.2 . Surviva


Respiratory viral illnesses are associated with diverse neurological complications, including acute transverse myelitis (ATM). Among the respiratory viral pathogens, the Coronaviridae family and its genera coronaviruses have been implicated as having neurotropic and neuroinvasive capabilities in human hosts.While there are currently no published reports on post-infectious ATM secondary to SARS-CoV-2, there is one report of parainfectious ATM attributed to SARS-CoV-2 in pre-print. Here, we present a case of infectious ATM attributed to SARS-CoV-2 in a 24-year-old male who presented with bilateral lower-extremity weakness and overflow urinary incontinence after confirmed SARS-CoV-2 infection. Magnetic resonance imaging revealed non-enhancing T2-weighted hyperintense signal abnormalities spanning from the seventh through the twelfth thoracic level consistent with acute myelitis.The patient underwent further workup and treatment with intravenous corticosteroids with improvement of symptoms and a discharge diagnosis of ATM secondary to SARS-CoV-2. Transverse myelitis is typified by an acute or subacute inflammatory myelopathy resulting in potentially disabling neurological deficits such as motor weakness and sensory deficits, as well as autonomic dysfunction. Acute transverse myelitis (ATM) has been associated with a variety of different etiologies, which have been subdivided into compressive and non-compressive myelopathies. Non-compressive etiologies include infectious, autoimmune, ischemic, paraneoplastic, radiation effects, post-vaccination, and post-infectious, as well as idiopathic causes. The diagnosis of ATM, while challenging for the clinician, is important to recognize as it is associated with significant morbidity and mortality leaving two-thirds of infected patients with moderate to severe permanent disability.4The incidence rate of ATM has been estimated to be between one and eight new cases per million annually, but recent data show that it may be as high as 31 cases per million.A 24-year-old male with no significant medical history presented to the emergency department (ED) with complaint of fever and chills, along with nausea and non-bloody, non-bilious vomiting. He denied recent travel or sick contacts. He was febrile and tachypneic with findings of patchy airspace disease throughout both lungs compatible with multifocal pneumonia on computed tomography of the chest without contrast. He was subsequently admitted to the hospital for three days. Nasopharyngeal swabs were positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) on reverse-transcriptase polymerase chain reaction (RT-PCR). He was treated with supportive care and demonstrated clinical improvement.His respiratory pathogen panel did not reveal any coinfection and his legionella antigen, blood cultures, respiratory cultures, and human immunodeficiency virus (HIV) testing were also negative. He was discharged home but subsequently presented to the ED nine days later with symptoms of bilateral lower-extremity weakness in addition to developing overflow urinary incontinence. He denied any history of trauma, pain, or similar symptoms in the past. Upon arrival, his vitals revealed a blood pressure of 111/61 millimeters of mercury, pulse 97 beats per minute, respiratory rate 16 breaths per minute, 98% oxygen saturation on room air, and temperature of 37.3° Celsius.His neurological examination revealed bilaterally absent knee and ankle reflexes with equivocal plantar reflexes, and flaccid, lower-extremity paraplegia bilaterally, in addition to evidence of overflow urinary incontinence. His lower-extremity sensory examination and anal tone were normal. His physical examination was otherwise normal. Laboratory workup included a complete blood count, complete metabolic panel, thyroid testing, inflammatory markers, repeat nasopharyngeal RT-PCR SARS-CoV-2 testing, and urinalysis, which were found to be unexceptional with a negative SARS-CoV-2 result on hospital day one. The patient underwent MRI of the spine as well as a lumbar puncture (LP). The MRI showed a non-enhancing T2-weighted hyperintense signal abnormality spanning from the seventh through the twelfth thoracic level consistent with acute myelitis .What do we already know about this clinical entity?Transverse myelitis is a focal inflammatory myelopathy causing motor, sensory, and autonomic dysfunction. Diagnosis rests upon clinical findings as well as serologic, magentic resonant imaging, and cerebral spinal fluid studies.What makes this presentation of disease reportable?Coronaviridae have been shown to have neurotropic and neuro-invasive capabilities, yet little is known about severe acute respiratory coronavirus 2 (SARS-CoV-2). We present the second case of acute myelitis attributed to SARS-CoV-2.What is the major learning point?SARS-CoV-2 is associated with a variety of neurological manifestations, including myelitis. Diagnosis should utilize established diagnostic criteria.How might this improve emergency medicine practice?Reinforcing the unique presentations of SARS-CoV-2 and myelitis yields a better understanding of the disease entities, allowing a focused investigation for accurate diagnosis and treatment.CSF studies from the LP were consistent with a lymphocytic pleocytosis, normal glucose and protein levels, and electrophoresis. The patient underwent further workup to rule out other causes of transverse myelitis with CSF immunoglobulin G index, CSF-specific oligoclonal bands, aquaporin-4 antibodies, B-12 level, methylmalonic acid level, as well as a workup for HIV, other infectious diseases, autoimmune disease, connective tissue disease, and multiple sclerosis. He was diagnosed with post-infectious myelitis secondary to SARS-CoV-2 infection. Treatment was initiated with intravenous (IV) methylprednisolone and supportive care with noted improvement in bilateral lower-extremity strength. Interestingly, repeat SARS-CoV-2 testing was done on hospital day four, which returned positive. Hence, it is difficult to ascertain whether the patient demonstrated post-infectious ATM as opposed to parainfectious ATM secondary to SARS-CoV-2.Coronaviridae family and its genera coronaviruses have been implicated as having neurotropic and neuroinvasive capabilities in human hosts.8ATM is characterized by focal inflammation of the spinal cord leading to varied severity of motor, sensory, and autonomic dysfunction. Although uncommon, it is paramount to distinguish it from other neurologic etiologies due to its potential for permanent disability. The diagnosis is based on characteristic clinical findings in addition to serologic, MRI, and CSF studies. As previously noted, the Previous studies in mice have proposed that human coronavirus may reach the CNS via the olfactory bulbs, as viral antigens were initially detected there followed by propagation and detection in whole brain tissue days later.11Recently, there has been a growing body of evidence supporting the association of SARS-CoV-2 with neurological abnormalities. A systematic review looking at the incidence of secondary neurological disease in patients diagnosed with SARS-CoV-2 found rates to vary from 6–36.4%.During his workup for ATM, this patient tested negative on repeat SARS-CoV-2 testing on hospital day one but tested positive for SARS-CoV-2 on hospital day four. Hence, it is difficult to ascertain whether the patient demonstrated post-infectious ATM as opposed to parainfectious ATM secondary to SARS-CoV-2. The diagnosis of parainfectious or post-infectious ATM relies upon a stepwise approach to rule out compressive etiologies as well as other inflammatory and non-inflammatory etiologies of ATM .Our patient met the inclusion criteria for diagnosis of ATM based on bilateral motor symptoms and autonomic dysfunction with bladder incontinence along with evidence of CSF lymphocytic pleocytosis and characteristic MRI findings while ruling out other infectious, autoimmune, and connective tissue etiologies. Treatment of ATM must be individualized to the patient and underlying etiology that may have caused ATM. There are currently no established regimens for treatment of SARS-CoV-2 post-infectious or parainfectious transverse myelitis. Treatment for other infectious-mediated ATM include antivirals, antibiotics, corticosteroids, and IV immunoglobulin, but their efficacy has yet to be completely defined. Overall, a single case report is not robust enough to suggest a definitive link between ATM and SARS-CoV-2. More research and case reports are necessary to support a causal relationship. Despite this, clinicians must be aware of the possibility of an association with SARS-CoV-2 and be aware of the salient features of ATM for early diagnosis, workup, and potential treatment to prevent permanent disability.In summary, we hypothesize that this patient’s ATM was precipitated by SARS-CoV-2 leading to a diagnosis of post-infectious or parainfectious ATM. ATM has a varied presentation and is associated with significant morbidity and mortality that necessitates increased awareness and vigilance on part of the clinician. This article is the second reported case of ATM attributed to SARS-CoV-2 infection, and should serve to reinforce the unique presentations of SARS-CoV-2 and ATM.


The pandemic severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been identified as the disease-causing pathogen of Coronavirus disease 2019 (COVID-19). (Pre)clinical research to identify rapidly available small molecules for the treatment of SARS-CoV-2 infections/COVID-19 has focused to date on the approved lysosomotropic antimalarials chloroquine and hydroxychloroquine, the investigational remdesivir , and the anti-inflammatory corticosteroid dexamethasone . LopinavDespite encouraging results in treating hospitalized patients with COVID-19 requiring supplemental oxygen, mechanical ventilation, or extracorporeal membrane oxygenation (ECMO) with remdesivir and dexamethasone, there is still a lack of active compounds exhibiting pan-coronavirus antiviral activity, tackling or preventing host cell infection, forming syncytia, endotheliitis, or the cytokine release syndrome (CRS)/cytokine storm syndrome in COVID-19. Target-oriented and in particular site of action-oriented drug repurposing of small molecules has the potential to close the gap in prophylaxis and treatment of mild and moderate COVID-19 and to reduce mortality in severe cases.Oxidative stress  has been demonstrated in animal models of SARS  and servSevere COVID-19 is associated with an atypical diffuse alveolar damage, ending in the acute respiratory distress syndrome (ARDS) , most li18-ceramide, are present in (sepsis-induced) cardiac dysfunction  mammalian cells is characterized by an increase in Cceramide  and can ceramide . Further as well .According to current knowledge, in therapy inhibition of lysosomal pH dependent processes  can be obtained only through off-label use of lysosomotropic drugs. Systemic application in lysosomotropic drug concentrations and obtaining an efficacious blood level is sometimes accompanied by severe adverse effects and/or (in this case) undesirable (intrinsic) pharmacological effects. Chloroquine was among the first lysosomotropic active compounds exerting antiviral effects on SARS-CoV-2  and duriin vitro (NCT04366739). Consequently, chlorpromazine is rated as a promising candidate in COVID-19/CRS treatment. In case of treatment of people without mental illness, however, a premature termination of treatment due to severe side effects by systemic application of chlorpromazine is extremely likely. This raises the question of how to handle this issue to provide well tolerated lysosomotropic drugs in SARS-CoV-2 infection/COVID-19.Chlorpromazine displayed anti-SARS-CoV-2 effects in vitro  and protNumerous available approved drugs with lysosomotropic characteristics permit tailor-made therapy. The individual pre-existing conditions are a criterion for the selection and combination of lysosomotropic drugs. For choosing suitable lysosomotropic drugs some issues have to be considered:Various lysosmotropic drugs in in vitro anti-SARS-CoV-2 antibiotics teicoplanin, oritavancin, dalbavancin, and telavancin  and fluoxetine (NCT04377308) as lysosomotropic drugs may provoke severe and unfavorable adverse effects in mental healthy patients. Since the respiratory tract is both, the gateway for SARS-CoV-2 infection/COVID-19 and an internal surface, the expedient is a local application in the airways and/or the respiratory tract. Local application of small molecules is possible, preferably as inhalant or via nebulizers to avoid (undesirable) systemic effects. The majority of lysosomotropic drugs should be suitable for inhalation.Systemic application of chlorpromazine   and virain vitro) (in vivo as demonstrated in vitro due to poor accumulation in pulmonary tissue (Nafamostat is an approved protease inhibitor that inhibits TMPRSS2 (n vitro) , preventn vitro) , appearsn vitro) . It is dn vitro)  is suffiy tissue .Various clinical trials are currently under way using immunomodulatory IL-1 and IL-6 inhibitors or anti-IL-6R antibodies  in patients with COVID-19 ; limitedTo improve outcome, antibody cocktails consisting of anti-IL-6, IL-1 receptor blocker, IL-1 type 1 receptor, and TNF-α are suggested , irrespeAlthough lysosomotropic drugs likewise interfere with the immune defense, such adverse effects are not reported. In contrast to antibodies, however, only the resynthesis of IL-6 and thus the available amount is reduced, but not completely obstructed, still allowing a moderate immune response. Multitargeting on core processes of the viral infection addressing the formation of multinucleate syncytia and alteration of tissue structure, ceramide metabolism, and the release of virions could be a key advantage of lysosomotropic drugs compared to current strategies.Daunting results of (hydroxy)chloroquine in clinical trials are closely related to their drug profile and minor lysosomotropism, but not to the mode of action (lysosomotropism) in general. Observations in patients treated with chlorpromazine and the extensive accumulation of imipramine in alveolar macrophages and of both, imipramine and chlorpromazine in isolated perfused lung tissue supports the benefits of lysosomotropic drugs that are accumulating in pulmonary tissue in SARS-CoV-2 infection/COVID-19.Promising candidates among lysosomotropic drugs in fact require more than adequate lysosomotropism; accumulation in pulmonary tissue is a prerequisite as well. It is, however, likely irrelevant whether the drug or its metabolite(s) is accumulating given the broad structural requirements for this activity. Since a large number of compounds has not yet been evaluated for lysosomotropism, many compounds beside those listed in


The global immuno-oncology pipeline has grown progressively in recent years, leading cancer immunotherapy to become one of the main issues of the healthcare industry. Despite their success in the treatment of several malignancies, immune checkpoint inhibitors (ICIs) perform poorly in others. Again, ICIs action depends on such a multitude of clinico-pathological features, that the attempt to predict responders/long-responders with ad-hoc built immunograms revealed to be quite complex. In this landscape, the role of nuclear medicine might be crucial, with first interesting evidences coming from small case series and pre-clinical studies. Positron-emission tomography (PET) techniques provide functional information having a predictive and/or prognostic value in patients treated with ICIs or adoptive T-cell therapy. Recently, a characterization of the tumor immune microenvironment (TiME) pattern itself has been shown to be feasible through the use of different radioactive tracers or image algorithms, thus adding knowledge about tumor heterogeneity. Finally, nuclear medicine exams permit an early detection of immune-related adverse events (irAEs), with on-going clinical trials investigating their correlation with patients’ outcome. This review depicts the recent advances in molecular imaging both in terms of non-invasive diagnosis of TiME properties and benefit prediction from immunotherapeutic agents. The host immune system interacts with tumor cells through the activation of innate and adaptive immune mechanisms. Initially, transformed cells are eliminated by a competent immune system. Nevertheless, sporadic tumor cells manage to survive immune destruction and may enter an equilibrium phase during which editing occurs. Finally, immunologically sculpted tumors begin to grow progressively, establish an immunosuppressive tumor microenvironment and become clinically apparent . The immThe global I-O pipeline has grown progressively in last years. The administration of interleukin-2 (IL-2) and the adoptive transfer of antitumor T cells grown in IL-2 represented the first effective immunotherapies for cancer in humans . RecombiMoreover, the development of immune checkpoint inhibitors (ICIs) is a revolutionary milestone in the field of I-O. ICIs reinvigorate antitumor immune responses by interrupting co-inhibitory signaling pathways and promote immune-mediated elimination of tumor cells. Anti-programmed-death 1 (PD-1) nivolumab and pembrolizumab, anti-programmed death-ligand 1 (PD-L1) atezolizumab, durvalumab and avelumab, and anti-cytotoxic T-lymphocyte antigen 4 (CTLA-4) ipilimumab are the standard of care in adjuvant treatment or advanced disease treatment for many solid tumors .However, despite these substantial advances, only a subset of patients receiving ICIs derive clinical benefit. It is then crucial to identify and to develop predictive biomarkers of ICIs response. PD-L1 expression and tumor mutation burden are the only predictive factors validated in phase III clinical trials, but are still imperfect since there are ICI non-responders expressing high biomarkers levels and ICI responders with low biomarkers levels. New determinants of response are being investigated, with strategies encompassing multiple biomarkers ,50.Nuclear medicine is a branch of medicine using radionuclides in the diagnosis and treatment of diseases. Diagnostic applications consist in “functional imaging” and are based on the ability of radiopharmaceutical agents to concentrate in pathological tissues and to emit radiations that are revealed by external detectors and then recorded. Therapeutic approaches exploit selective concentration of radiopharmaceutical agents in pathological tissues and the use of the radiations to destroy them. Techniques of nuclear medicine imaging include two-dimensional scintigraphy, three-dimensional single-photon emission computed tomography (SPECT) and positron emission tomography (PET), and hybrid techniques combining computed tomography (CT) or magnetic resonance imaging (MRI) scans with SPECT or PET .18F]fluoro-2-deoxy-d-glucose (18F-FDG or FDG) PET/CT has become an established standard nuclear imaging modality in oncology [2-[oncology  based ononcology . FDG-PEToncology ,55,56.However, some drawbacks have to be mentioned . InfectiAssessment of response represents a measure of antitumor drug activity. Radiological overall response rate is the primary endpoint of about 70% of phase II trials. Thus, positive trials are the basis of accelerated approval in refractory advanced disease, a setting where there is no available therapy . FurtherPractically, response assessment starts with summing the size of tumor lesions in a baseline CT scan before beginning a new therapy. After a specified time, the relative change of the sum of lesion sizes is registered. Finally, a threshold on this relative change is applied to decide whether treatment should be administered or discontinued. Historically, the WHO Response Criteria were first published in 1981 and were based on bidimensional measurements . As CT sNevertheless, less than half of the studies that met their response endpoint measured with RECIST criteria in a phase II clinical trial are successful in phase III clinical trials ,63,64. ITwo metabolic response criteria were thus developed. European Organization for Research and Treatment of Cancer (EORTC) PET response criteria were based on standalone PET imaging and focused on evaluation of only one lesion . PET ResResponse assessment for I-O must deal with a unique challenge, pseudoprogression, that is a temporary increase in tumor size, meeting the criteria for progressive disease based on RECIST, which is lately not confirmed. Pseudoprogression is caused by immune cells infiltrating tumor lesions. Its prevalence is up to 10%, and it has been characterized in malignant melanoma studies ,70.Four new response criteria have been proposed to solve this issue and all of these require a confirmation of progressive disease . Immune-IrRC and irRECIST were clinically validated in patients with advanced malignant melanoma treated with ipilimumab, imRECIST in patients with advanced NSCLC and metastatic urothelial carcinoma treated with atezolizumab, whereas iRECIST are based on a consensus statement. The RECIST working group recommends that recent trials should use RECIST 1.1 to define the primary and secondary efficacy-based endpoints and reserve irRC or irRECIST for exploratory endpoints .The role of already published metabolic criteria was investigated in patients treated with immunotherapy. EORTC criteria applied to PET/CT evaluation after two cycles of ipilimumab on 22 advanced melanoma patients were found to be predictive of final treatment outcome . RetrospLikewise, semi quantitative parameters were found to be predictive of outcome in patients treated with immunotherapy. This could be the case for SUV, metabolic tumor volume (MTV), and tumor total lesion glycolysis (TLG). These three variables define the entire metabolic tumor burden (MTB), that may represent a reliable value to predict treatment success, being known that I-O work better in a context of low tumor load . VariatiIn parallel, several I-O-adapted metabolic criteria have been proposed. PET/CT Criteria for Early Prediction of Response to Immune Checkpoint Inhibitor Therapy (PECRIT) are based on RECIST 1.1 to define complete response, partial response and progressive disease, whereas percent change in SULpeak by PERCIST criteria is used to distinguish patients with stable disease per RECIST 1.1 and clinical benefit from those with stable disease and no clinical benefit. The comparison of baseline PET/CT scan with a second PET/CT scan at 3–4 weeks after the start of immunotherapy based on PECRIT was able to predict the best overall response at 4 months in 20 patients with advanced melanoma patients treated with nivolumab, ipilimumab or BMS-936559 .Conversely, the number of new FDG-avid lesions was considered in PET Response Evaluation Criteria for Immunotherapy (PERCIMT). Four newly emerged avid lesions on PET/CT scan after 3 months of therapy was found to be predictive of treatment failure in 41 melanoma patients treated with ipilimumab, with the functional size of the new lesions playing an important role in predicting the clinical response . PERCIMTInflammatory response caused by immunotherapy may confound the ability of FDG-PET to identify patients with a favorable response. Thus, similarly to iRECIST, immune PET Response Criteria in Solid Tumors (iPERCIST) included the concept of disease progression confirmation in PERCIST. iPERCIST were retrospectively applied to a cohort of 28 NSCLC patients treated with nivolumab. All patients underwent a baseline PET/CT scan and a second scan after 2 months of treatment, which was evaluated with the proposed criteria. A metabolic unconfirmed progressive disease was re-evaluated with a third PET/CT scan after 4 weeks. The 1-year survival rates were significantly higher in iPERCIST responders compared with non-responders . Beyond The survival benefit from using ICIs varies considerably from patient to patient. ICIs manage to slow down cancer progression, changing completely long-term survival perspective for a subgroup of patients, that goes from 10% to 30%, depending from the tumor types and drug used ,91. The Last year, scientific community efforts in oncology were greatly conveyed to unhinge the reasons of this imbalance, trying to recognize the ideal candidate for immunotherapy; the final aim is to sharpen and potentiate ICIs tools . ICIs acThe first studied marker has been PD-L1, a protein variously expressed by tumor cells, that for some cancer types correlate well with benefits from specific anti-PD-L1 agents ; in factEvaluating US Food and Drug Administration (FDA) approvals of 45 ICIs agents across 15 tumor types, PD-L1 was found to be predictive in less than a third of them . This phThus, it would be of extreme importance to consider TiME globally in patients eligible for ICIs. Analyses of TiME on tissue could be affected both by tumor tissue sample itself in terms of quantity and/or quality and by tumor heterogeneity . NM imag68Gallium-labeled chelator-linked FAP inhibitors (FAPIs) and 18F-labeled glycosylated FAPIs [Recently, imaging of TiME by PET has gained an excellent progress by the introduction of different tracers targeting fibroblast activation protein (FAP), among which ed FAPIs . FAP is ed FAPIs . NM imaging can provide a static baseline representation of the identified marker, so having a predictive value, possibly bringing our therapeutic choices to be as careful as possible. Otherwise, these techniques can be used in a more dynamic way, by repeating NM imaging in different disease time-points in patients treated with ICIs. The aim will be to monitor treatment response, by using labeled-radiotracers conjugated with immunotherapeutic agents, with first brilliant results in this sense.89Zr-desferrioxamine-labeled cys-diabody permitted a noninvasive immuno-PET tracking of endogenous CD8+ T cells in melanoma immunotherapy models, including anti-PD-L1 agents, antigen-specific adoptive T-cell transfer and agonistic antibody therapy.The generation of anti-CD8 The great power of such imaging consists in its ability to acquire dynamic information regarding therapy-induced alterations of both PBMCs and TILs component .89Zr)-labeled PEGylated single-domain antibody fragments specific for CD8, Rashidian et al. demonstrated the ability to longitudinally detect TILs in melanoma models and to distinguish responding tumors from those that do not respond to anti-CTLA4 therapy [Similarly, with an immuno-PET with Zirconium-89( therapy .123I-interleukin-2 (IL-2) single-photon emission computed tomography (SPECT) well depicted TILs in cancer patients, also giving a preliminary response to cytokine treatment [99mTc-IL-2 in melanoma patients treated with I-O permits to acquire information about pseudo-progression and prognosis [89Zr [64Cu-DOTA-anti-CTLA-4 mAb [64Cu or 89Zr can detect very precisely PD-1-expressing TILs [89Zr using the bifunctional chelator p-SCN-Bn-Deferoxamine (DFO) led to an immune-PET detection of LAG-3 expressing TILs [reatment ,104, whirognosis ,106. Prois [89Zr . RecogniLA-4 mAb ,109, whiing TILs ,112,113.ing TILs . NM indi68Ga-labeled single-domain antibody tracer called 68Ga-NOTA-Nb109, and 89Zr-DFO-PD-L1 monoclonal antibody [89Zr was demonstrated in tumors responding to DS-5573a therapy [89Zr-DFO-daratumumab provides successful whole-body PET visualization of myeloma, and predicts the effectiveness of daratumumab therapy [A similar usefulness may derive from PET-imaging of some proteins expressed directly by tumor cells. High affinity to PD-L1 has been guaranteed by specifically synthetized antibody ,116. Mor therapy . 89Zr-DF therapy .All these applications represent in a certain way an evolution of the most widely used FDG-PET, already proved as seen to acquire response to ICIs earlier than through the classical CT scan imaging ,78,79.The therapeutic approach cannot ignore inter-patients and intra-patient cancer heterogeneity. The latter depends on a complex phenotypic evolution that involve (epi)genetic transformations, surface markers selections and cell-cycle modifications in response to environmental stimuli . What is18F-FDG PET imaging can point out the presence of aggressive tumor foci, which may divert patients’ prognosis [18F-FDG PET and to enhance tumor uptake [NM may be helpful to visualize tumor heterogeneity, with interesting applications in the field of I-O. Quantitative analyses of canonical rognosis ; CAFs har uptake .The recognition of TiME and cancer cells markers heterogeneity, through specific tracers, could guide in the future daily therapeutic choices. PD-L1 is the forerunner marker in this sense, with different contributes about its possible discordant expression in different tumor lesions ,125,126,Immune-related adverse events (irAEs) are unique adverse events caused by activation of an immune response against healthy tissues due to immunotherapy. IrAEs occur in about two thirds of patients treated with IO, of which 14% is grade 3 or superior . Timely Diagnosis of irAEs is made through clinical, biohumoral, and radiological investigations. Early radiological detection of irAEs could improve their correct management. In a recent study, medical imaging detected 74% of irAE in patients treated with anti-PD1. Among typical irAEs, interstitial lung disease is mainly diagnosed with CT scans, as well as enterocolitis and pancreatitis, whereas thyroiditis with ultrasound and hypophysitis with magnetic resonance imaging .Beyond tumor lesions, FDG uptake can be induced by the inflammatory reaction related to irAEs. Thus, sarcoid-like reaction ,133 and Furthermore, irAEs could predict response to immunotherapy . A retroNM imaging has historically contributed in a decisive way to expand the knowledge about cancer management and cancer response to treatment, providing several functional data. In the context of modern I-O, this could become quite challenging, because each day millions of cancer patients worldwide are treated with I-O therapies, and lots of crucial aspects remain to be clarified. Last decade scientific reports added progressively important pieces to the definition of the complex immune-cancer cycle, finding out favorable clinical and pathological patterns of response to I-O drugs, and even creating complex predictive algorithms.Main open questions include the optimal candidate for I-O therapy, the determination of response and appropriate duration of treatments, the mechanisms that underlie primary and secondary resistance to I-O. The goals are I-O empowerment in terms of efficacy and healthcare resources optimization.As seen, PET-CT scans has shown to predict responders and non-responders to I-O earlier than the conventional CT scan, through the acquisition of both anatomical and metabolic data. Moreover, several PET-CT scan parameters, acquired at baseline or during treatment, have been found to be predictive of response to I-O. NM applications could well depict several predictive TiME features by using different radiotracers and/or drug-radiotracer conjugates, and also precociously recognize drug-induced irAEs usually correlated with I-O benefit. This would permit ideally to change in the near future the approach of cancer care, by selecting the right I-O drug for the right patient, revealing the immunologic reasons of innate and acquired resistance to I-O, and laying the foundations to get around them.In the last few years, I-O imaging field has been enriched by radiomics, a set of methods used to extract a large amount of quantitative data from biomedical images—called features—aiming to personalize medicine even more. The typical radiomics workflow is a multi-step process, that starts with the acquisition of measurable features and, through different phases of data processing, concludes with data analysis . Radiomi


A 37-year-old Burmese woman presented with an incidentally found retroperitoneal fat-containing tumor. The tumor was 9 cm in the longest diameter, surrounding the right kidney, and composed of homogenous adipose tissue with thickened septum-like structures and spotty nonadipose structures, which were enhanced on contrast-enhanced computed tomography and magnetic resonance imaging. The tumor did not show either a beak sign or synchronous angiomyolipoma-like lesion in the kidneys. The tumor had irregular septa, thin blood vessels, and spotty small soft-tissue nodules. The tumor did not contain any heterogeneously enhanced solid lesions suspicious for dedifferentiated liposarcomas. Based on these imaging findings, a clinical diagnosis of a well-differentiated liposarcoma was made. Under the consensus of a multidisciplinary cancer board, she was recommended to undergo core-needle biopsy to confirm the clinical diagnosis. However, she declined to undergo biopsy for financial reasons. She underwent kidney-sparing retroperitoneal tumor resection. Histopathologically, the tumor was an angiomyolipoma with positive immunostaining for HMB45 and Melan A. The present case suggests the importance of core-needle biopsy prior to surgical intervention for retroperitoneal fat-containing tumors. Several types of fat-containing tumors arise in the retroperitoneum. Of these, the differential diagnosis between well-differentiated liposarcomas (WDLs) and large renal angiomyolipomas (AMLs) is sometimes challenging , 2. AlthA 37-year-old Burmese woman presented with an incidental retroperitoneal tumor. Contrast-enhanced computed tomography (CT) revealed a tumor measuring 9 cm in the longest diameter surrounding the right kidney . The tumWe encountered a case of a renal AML mimicking a WDL. It is occasionally challenging to differentiate AMLs from WDLs because both tumors might appear as similar large fat-containing perinephric masses. To distinguish them radiologically, it has been reported that renal AMLs often exhibit characteristic findings of a “beak sign” and “embedded organ sign” on CT and MRI . AdditioWDLs and DDLs are known to originate from retroperitoneal soft tissue and harbor adipose tissue and thickened irregular septum-like structures exhibiting strong enhancement , 8. AlthWe encountered a case of a renal AML mimicking a WDL on imaging examinations. The important role of core-needle biopsy is suggested for accurate clinical diagnosis and proper management of retroperitoneal fat-containing tumors.


Numerous potyvirus studies, including virus biology, transmission, viral protein function, as well as virus–host interaction, have greatly benefited from the utilization of reverse genetic techniques. Reverse genetics of RNA viruses refers to the manipulation of viral genomes, transfection of the modified cDNAs into cells, and the production of live infectious progenies, either wild-type or mutated. Reverse genetic technology provides an opportunity of developing potyviruses into vectors for improving agronomic traits in plants, as a reporter system for tracking virus infection in hosts or a production system for target proteins. Therefore, this review provides an overview on the breakthroughs achieved in potyvirus research through the implementation of reverse genetic systems. Reverse genetics of RNA viruses refers to the generation of recombinant viruses through site-directed mutagenesis such as substitution, deletion or insertion . This coPotyvirus is a genus of virus that belongs to the family Potyviridae. Potyviruses are among the most economically important and widely spread groups of plant viruses [Potato virus Y as the type species [ viruses . The gen species . The pic species  is trans species . Potyvir species . The mem species . The sin species ,16 gives species ,18. Addi species ,20.Brome mosaic virus [Cauliflower mosaic virus 35S promoter in a binary vector. Cells transformed with the plasmids containing virus cDNA clones are then introduced into plants through agroinfiltration, particle bombardment, or rubbing onto the leaf’s surface [Infectious clone technology represents the most commonly applied reverse genetics system. Cloning of an infectious clone was first described for ic virus . Since tic virus ,23. On t surface ,25. In tPoint mutation refers to a change occurred within a gene due to a single base pair alteration in the DNA sequence. During translation, conversion of RNA copied from the DNA into a sequence of amino acids would take place and point mutations usually cause a variety of effects on the protein synthesized at the final stage .Potato virus A (PVA). However, aphid transmissibility of the virus was still retained [Clover yellow vein virus, pClYVV cDNA clones containing no poly(A), poly(A) with shorter A residues, or various oligonucleotide sequences downstream were constructed. All RNA progenies obtained from the pClYVV cDNA constructs had poly(A) tails with infectivity varies along the sequences introduced. This suggested the addition of poly(A) to be independent of template, yet essential to maintain the infectivity of potyviral cDNA [Plum pox virus (PPV) revealed that the region between nucleotides 39 and 145 contributes to competitive fitness of the viral population [Turnip mosaic virus, TuMV-pXBS7/TuMV-pXBS8 chimeric viruses have allowed the mapping of a symptom severity determinant in the 3′-terminal UTR of pXBS8-derived virus [Mutagenesis studies aimed to characterize the phenotypic consequences resulting from genome modification, based on how the progenies differ from their respective wild type . To dateretained . Furtherral cDNA . Besidespulation . A pointed virus .Papaya ringspot virus (PRSV) were changed to aspartic acid and lysine respectively. Consequently, PRSV mild mutants with a potential to cross-protect Cucumis melo plants against wild type PRSV-W were obtained [Bean yellow mosaic virus (BYMV) was generated by exchanging an amino acid within the large ORF, leucine with serine. BYMV-M11 mutant conferred a complete cross protection against BYMV isolates from gladiolus, incomplete against BYMV isolates from other hosts, partial against a CIYVV isolate [Zucchini yellow mosaic virus (ZYMV) HC-Pro were substituted with isoleucine and asparagine respectively. The attenuated mutants were found to induce only mild symptoms with recovering and protected squash plants completely against the severe Taiwan strain, ZYMV TW-TN3 [Cross protection is conferred by pre-infecting the host crops with a mild virus strain to prevent the infection by another severe or closely-related strain of that virus . This stobtained . Similar isolate . An argiV TW-TN3 . AnotherV TW-TN3 .Mutagenesis studies on potyvirus infectious clones allowed the mapping of viral determinants responsible for viral genome replication, local and systemic movement of virus, symptomatology and host species range  Table 2Table 2. Potato virus Y, PVY-SON41 was demonstrated as the avirulence factor corresponding to Pvr resistance gene in pepper. A single substitution of adenosine nucleotide to guanosine at position 8424 in that region is sufficient for the virulence [The differential infectivity of UK 1 and JPN 1 strains of TuMV in Ethiopian mustard was studied to map the viral determinants involved . Isolateirulence .Brassica oleracea L.). Meanwhile, Tu-3 induced systemic chlorotic and ringspot symptoms in infected cabbage but not radish. The genomic region encoding C terminus of HC-Pro, all of P3 and N terminus of 6K1 was exchanged between pTuC and pTuR1 chimeras. The only difference between pTuR1 and pTuC was identified within the amino acid sequence of P3 gene. Hence, P3 gene was suggested to be involved in the differential phenotypes caused by TuMV isolates in both hosts.The differential responses of UK 1 and JPN 1 strains of TuMV in different hosts; brassicas  and radish cultivars , Daikon) were investigated . AlthougJohnsongrass mosaic virus (JGMV) strains, Johnsongrass infecting JGMV-Jg and Krish strain JGMV-Kr, was studied. Transcripts of JGMV-Jg full-length chimera containing coat protein sequences from JGMV-Kr were infectious in Krish resistant sorghums, thus confirming the role of JGMV-Kr coat protein in its host specificity [Chenopodiaceae and Cucuribitaceae families whereas PRSV P infects plants in papaya family (Caricaceae) additionally [Host specificity of two different cificity . Similartionally . Host astionally . A singltionally .Bean common mosaic virus (BCMV) interaction with its host, bean genotypes carrying different combinations of resistance genes . The experimental plants were agroinoculated with an infectious clone of RU1 strain, pCA-RU1-GUS containing UidA gene. In situ histochemical GUS assays revealed that DW, The Prince, CRM, SP, and SGR cultivars (carrying bc-u resistance gene) were all systemically infected. However, the genotypes carrying the I gene alone (Widusa) or in combination with bc-1 , bc-12/bc-22 (IVT-7233), and bc-3  showed only a weak blue staining. These resistance responses against BCMV-RU1 are largely in agreement with previous results obtained through immunological method and symptom descriptions based analysis [I conferred a complete protection against BCMV-RU1 while bc-genes do not provide any. Besides that, the host defence response of cucurbit genotype, Dina-1 against ZYMV-NAA potyvirus has also been focused on previously [Naderpour and Johansen  studied analysis . The stueviously . Switche2 vesicles too. TuMV 6K2 fused to photoactivable GFP (PAGFP) was used to monitor the vesicle movement. Viral egress is shown to begin with the budding of 6K2 vesicles at ER export site in the globular structure. The virus then travels to plasma membrane and plasmodesmata to be delivered into neighboring cells. Some peripheral vesicles were also observed to be recycled back to the globular structure. This indicated a functional linkage between the peripheral vesicles and perinuclear structure. Although the Golgi apparatus and ER lost their organization due to the amalgamation, they were still connected to the host secretory pathway (ER-to-Golgi transport). The importance of this connection was investigated by inhibiting the pathway. As a result, the disruption enhanced the clustering of peripheral 6K2 vesicles with COPII coatamers, leading to an inhibition of cell-to-cell movement of virus. This suggests the requirement of a functional secretory pathway for a successful intercellular propagation of TuMV.To learn more on mechanisms of interrelation between potyvirus and host, the impact induced by TuMV infection on the endomembranes of host early secretory pathway was examined . TuMV inTobacco etch virus (TEV) and Arabidopsis thaliana proteins was also investigated [Arabidopsis thaliana proteins targeted by viral proteins were identified through affinity purification followed by mass spectrometry analysis (AP-MS). VPg and NIaPro specifically targeted 89 and 76 of these proteins, respectively. Overall, a total of 67 proteins targeted by both domains were considered to be the targets of full-length NIa.Along with these host-viral factor interactions, the interplay between stigated . An affiApart from the mature proteins, proteolytic processing of the potyviral polyprotein also produced multiple partially processed intermediates . The difTobacco vein mottling virus, TVMV-WT. The mutation suggested might have altered the motif structure and thus the binding property of HC-Pro too [r lacking 207 nucleotides in HC-Pro sequence exhibited an aphid-non-transmissible phenotype. However, its transmission activity was restored partially by pre-feeding the aphids on active HC-Pro from PVY. This confirms the helper activity of the N-terminal domain of HC-Pro [Potyviral HC-Pro and CP proteins were reported to have a crucial role in efficient aphid transmission . In line-Pro too . Likewisf HC-Pro . A mutatf HC-Pro . The motf HC-Pro .Soybean mosaic virus, SMV-G7H. A highly conserved histidine in the CP C-terminus and an arginine near the cleavage site at HC-Pro C-terminus were mutated and the results obtained showed that both amino acids are necessary to maintain the interaction for a successful transmission of SMV by aphids. Moreover, an amino acid substitution in the DAG motif was found to have disrupted the CP–HC-Pro interaction in YTHS.Seo et al.  conducteTobacco vein banding mosaic virus, TVBMV-HN39 infectious clone, pCaTVBMV-GFP were deleted. As a consequence, the mutant caused no systemic infection in inoculated Nicotiana benthamiana plants. According to the RNA secondary structures prediction, the deleted region is able to form a stem-loop (SL) like structure. Progenies derived from TVBMV mutants lacking nucleotides between positions 1 and 20 and 15 and 35 within 3′-UTR were found to have restored the 5′-end SL like structures and systemically infected tobacco plants. Hence, the 5′-terminal stem loop was proposed to be neccessary for TVBMV systemic infection [Clover yellow vein virus (CYVV-NZ) has been reported previously [In order to figure out the function of 3′-UTRs of potyviruses, the sequences between nucleotide positions 8–42 in the 3′ UTR of a nfection . Formatieviously .Apart from bacterial and yeast expression systems, the plant viral vectors also provide a fast and efficient approach for synthesis of specific proteins in plant cells . In thisStrawberry vein banding virus (SVBV) viral promoter was inoculated into experimental host plants [Viral vectors are usually tested through an expression of well-analyzable reporter genes in plants . With ret plants  (Table 3t plants  (Table 3Brome mosaic virus (BMV) to be developed into a plant virus vector was successfully demonstrated in 1986 [in planta (N. benthamiana using a Pepper mottle virus, PepMov-based vector led to an increased senescence along with milder symptoms [Rosea1 tagged infectious clone of PVY was developed, conferring benefits to molecular farming by rapidly produced larger amounts of anthocyanins in biofactory crops [The potential of  in 1986 . Since tn planta . The maisymptoms . Anothersymptoms . For an symptoms . Likewisry crops .Reverse genetics in virology relies on cDNA intermediates in order to genetically manipulate RNA viruses and further produce biologically active RNA molecules. Likewise, the available infectious full-length cDNA clones of potyviruses enable countless reverse genetics studies on phenotypic alteration and cross-protection by mutated viruses as well as in determining viral elements responsible for a particular biological characteristic of the virus. Apart from improving our understanding on the complexities of interactions between host and viral factors, reverse genetics strategies have also made various applications of recombinant vectors possible. Furthermore, the approach contributed immeasurably to the elucidation of basic functions of potyviral proteins, certain motifs or genome sequences in viral replication, transmission, and cell-to-cell movement. All these findings, in conjunction with different approaches such as transciptomics and proteomics analyses, would lead to the building of a larger network that helps to further explore potyviruses, especially in the identification of more durable resistance genes. Hence, reverse genetics technologies of potyviruses are believed to hold great promise for commercial applications in the future. Despite these breakthroughs, although various viral determinants have been identified through plant–potyvirus pathosystems, the host targets of those determinants are yet to be characterized in the future.


Traditional anti-arrhythmic drugs are classified by the Vaughan-Williams classification scheme based on their mechanisms of action, which includes effects on receptors and/or ion channels. Some known anti-arrhythmic drugs do not perfectly fit into this classification scheme. Other medications/molecules with established non-anti-arrhythmic indications have shown anti-arrhythmic properties worth exploring.In this narrative review, we discuss the molecular mechanisms and evidence base for the anti-arrhythmic properties of traditional non-antiarrhythmic drugs such as inhibitors of the renin angiotensin system (RAS), statins and polyunsaturated fatty acids (PUFAs).In summary, RAS antagonists, statins and PUFAs are ‘upstream target modulators’ that appear to have anti-arrhythmic roles. RAS blockers prevent the downstream arrhythmogenic effects of angiotensin II – the main effector peptide of RAS – and the angiotensin type 1 receptor. Statins have pleiotropic effects including anti-inflammatory, immunomodulatory, modulation of autonomic nervous system, anti-proliferative and anti-oxidant actions which appear to underlie their anti-arrhythmic properties. PUFAs have the ability to alter ion channel function and prevent excessive accumulation of calcium ions in cardiac myocytes, which might explain their benefits in certain arrhythmic conditions.Clearly, whilst a number of anti-arrhythmic drugs exist, there is still a need for randomised trials to establish whether additional agents, including those already in clinical use, have significant anti-arrhythmic effects. Th. Thphasediastole . Other cotential .Fig. 2Il2The Renin-Angiotensin System (RAS) is an established key player in the regulation of fluid and electrolyte balance and blood pressure. Angiotensin II (Ang II) is the main effector peptide of RAS, acting primarily via angiotensin II type 1 (AT1R) and 2 receptors (AT2R) . Angiote2.1Anti-arrhythmic roles for ACE-Is and AT1R blockers appear plausible on consideration of the molecular effects of some components of RAS. Chronic AF is associated with the down-regulation and up-regulation of AT1 and AT2 in human atrial tissue, respectively . SimilarRAS could also influence arrhythmogenicity via modulation of extracellular matrix protein expression and cardiac remodelling. Ang II leads to proliferation, while Ang-(1−7) leads to anti-proliferation. Progressive accumulation of fibrotic tissue in the myocardium is a major contributor to structural cardiac remodelling, along with dilatation and myocardial hypertrophy. Structural remodelling includes changes in both the cellular components  and the extracellular matrix. Ang II has direct proliferative effects on atrial and ventricular fibroblasts and smooth muscle cells . Ang II 2.2AF is the commonest type of arrhythmia; and it is associated with remodelling , which facilitate the occurrence of the arrhythmia. In many cases, the electrical remodelling is thought to be mediated by rate-induced intracellular calcium overload in the short term ,21, and INa) density and the atrial conduction velocity [INa by promoting the formation of hydrogen peroxide (H2O2), which instigates the binding of nuclear factor kappa B (NF-κB) to the SCN5A promoter [INa [Chronic rapid atrial activation (by pacing) promotes the induction of sustained AF. In a canine model, this was shown to occur from a reduction in both the sodium current  in AT1R knock-out mice was much less compared to the number in wild-type controls in ischaemia-reperfusion injury; and that selective treatment with an AT1R blocker in wild-type mice prior to ischaemia ameliorated the occurrence of the PVCs .Reduction in atrial fibrosis may also contribute to reduction in structural remodelling, and hence modulation of the pro-arrhythmic substrate. By virtue of their antifibrotic properties, RAS inhibitors are believed to play a role here by reducing the synthesis of collagen I molecules as well as promoting the degradation of collagen I fibres already formed , and red2.3Potential beneficial effect of RAS inhibition on arrhythmias (e.g. AF) could be direct or secondary to neurohumoral modulation and antihypertensive effects. RAS inhibition has also been associated with indirect anti-arrhythmic effects – as identified for some clinical trials set-up for non-arrhythmic primary outcomes . Randomised controlled trials are weightier measures on the evidence scale; hence, a few trials have sought to prove a direct anti-arrhythmic effect for RAS inhibitors with rather mixed results .Table 2RTRACE) study generated very promising results. Patients with reduced left ventricular function (LVF) secondary to acute myocardial infarction (AMI) in sinus rhythm (SR) received either trandolapril or placebo and were followed up for 2–4 years for the primary outcome measure of the development and time to occurrence of AF in one 12-lead ECG recorded at an outpatient visit. The study showed a significant difference in the development of AF in favour of the ACE-I with a relative risk of approximately 50 % albeit with a broad confidence interval) in patients with left ventricular dysfunction (LVD) secondary to AMI (95 % CI: 0.26−0.76 [GISSI-3) there was a 24 % reduction in AF incidence  in patients with both an ACE-I (lisinopril) and nitrates compared to controls [SOLVD) involving patients with LVD in SR revealed a significant difference in the incidence of AF in patients who had received enalapril compared to those who received placebo after mean follow up of nearly 3 years. On multivariate analysis, enalapril was the most powerful predictor for risk reduction of AF  [Val-HEFT) showed a significant difference in AF incidence in patients with HF randomised to either valsartan or placebo on top of HF treatment. [HOPE trial, however, found that over 4.5 years of follow-up ramipril (compared to placebo) did not significantly reduce the incidence of AF in patients without known HF or left ventricular systolic dysfunction (LVSD) [The TRAndolapril Cardiac Evaluation (.26−0.76 . In largcontrols . Furthercontrols . A furth11−0.44) . Similareatment. . A meta-eatment. . In contn (LVSD) . Studiesn (LVSD) ,43.GISSI-AF trial, valsartan was not associated with a reduction in the incidence of recurrent AF [ONTARGET, ACTIVE I, and J-RHYTHM II have found little additional beneficial effect against AF recurrence using RAS blockade along with an AAD  [However, a number of studies are less promising; in the rrent AF  and trearrent AF . Furtherafenone) .3Statins are important medications with established indications in primary and secondary prevention of coronary heart disease. They decrease the cellular cholesterol content by selectively inhibiting the rate-limiting enzyme of cholesterol synthesis pathway : 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase. As well as limiting cholesterol biosynthesis, they lower serum and hepatic cholesterol and serum triglyceride concentrations.3.1A counter-regulatory mechanism of reduced cholesterol biosynthesis is upregulation of low-density lipoprotein (LDL) receptor expression in the hepatocyte membrane, with accelerated hepatic clearance of circulating LDL cholesterol . FurtherBy reducing the production of mevalonate (a key intermediate compound during cholesterol biosynthesis) through the inhibition of HMG-CoA reductase, statins impair the downstream production of isoprenoid intermediates (e.g. farnesylpyrophosphate and gerany-geranylpyrophosphate). This implies inhibition of post-translational modifications , that hydrolyse guanosine triphosphate, i.e. GTPases ). Rho an-atherosclerotic plaque stabilisation and regression ,61, (i.e-inhibition of oxidised LDL (oxidised LDL upregulates AT1R possibly via a nuclear factor kappa B- NFκB-dependent pathway) ,63,-anti-oxidant, anti-inflammatory and anti-proliferative effects ,-regulation of nitric-oxide-dependent coronary arterial tone and endothelial function ,68,-attenuation of arrhythmogenic structural remodelling via mechanisms including upregulation of transcription factor GATA-6 expression and inhibition of the Rho pathway ,-improvement in the heterogeneity of cardiac repolarisation   and-changes in transmembrane ion channel conductance .Briefly, the possible underlying mechanisms of anti-arrhythmic effects of statins include:3.2MADIT-CRT  was a randomised trial that examined the potential anti-arrhythmic benefit of statin use in non-ischaemic cardiomyopathy (NICM). Statin use was associated with a significant (77 %) decrease in the risk of cardiac death or life-threatening ventricular arrhythmias (VT/VF) [MADIT-II trial also showed beneficial anti-arrhythmic effect of statin use in implantable cardioverter-defibrillator (ICD) patients by lowering the risk of VT/VF and cardiac death [AVID) trial [GRACE), there was association with reduced VT, VF or cardiac arrest (as well as AF) [TRACS) – associated statins with decreased incidence of ventricular arrhythmias (VA) [ (VT/VF) . Similarac death . Statin D) trial ; while il as AF) . Furtherias (VA) .In summary, statins appear to show beneficial protective effect against life-threatening ventricular arrhythmias. The possible mechanisms underlying this protective effect may be related to effects including improved coronary artery tone, atherosclerotic plaque stabilisation, anti-inflammatory, anti-oxidative, and improved cardiac repolarisation heterogeneity .3.2.1Inflammation and oxidative stress have been of key interest among the possible mechanisms that underlie AF pathogenesis. In acute inflammatory states, such as cardiac or non-cardiac surgery and myo-pericarditis, new-onset AF coincides with peak levels of inflammatory biomarkers . Theoret2 score ≥ 2 than CHADS2 = 1  [Statins decreased the risk of new-onset AF by 19 % - more so with CHADSatabase) . On the atabase) .A meta-analysis by Fang et al., showed an overall significant reduced risk of AF incidence/recurrence .Inflammation and abnormal oxidative stress may be the main pathophysiological features related to atrial remodelling and enhanced myocardial tissue inflammation, resulting in AF onset, recurrence, and persistence . Statins4Polyunsaturated fatty acids (PUFAs) are best known as constituents of fish oils, but they are also found in soybean and certain rapeseed oils but not ordinary olive oil . N-3 and4.12+ overload [Ito, Ik, voltage-dependent sodium current) [Experimental studies show numerous potential molecular mechanisms by which PUFAs could potentially alter cardiac rhythmicity. At the level of the heart itself, PUFA exposure can alter myocardial membrane phospholipid composition and can, in turn, be anti-inflammatory or decrease potential Caoverload . Either current) . Ca2+ recurrent)  and thercurrent) ,129.  linking reduced AF with high fish consumption, the clinical trials investigating whether n-3 PUFA protect against AF are again disappointing. In the large OPERA multicentre trial, fish oil failed to reduce post-operative AF  and a siIn summary there is still no consensus to what the anti-arrhythmic potential of PUFA really are, if any, despite approaching 40 years of investigation .5In summary, RAS blockers, statins and PUFAs appear to have beneficial roles in decreasing the incidence and recurrence of arrhythmias. The foregoing discussions illustrate that while some insight has been gained in the potential molecular pathways underlying their plausible anti-arrhythmic roles, further clarification of these mechanisms and additional direct evidence of antiarrhythmic effects through randomised controlled trials are necessary.Only a few randomised trials to date have examined the potential anti-arrhythmic role of RAS blockers with no clear significant beneficial effects. However, there might still be some justification for future attempts aiming to identify direct anti-arrhythmic evidence through larger randomised controlled trials in patients with or without underlying cardiac disease. Future studies with statins could attempt to elucidate further the underlying molecular pathways explaining their pleiotropic effects. Furthermore, the identification of direct evidence for their anti-arrhythmic beneficial effects and the specific underlying pleiotropic effect at play are necessary. Further research into the differing effects of the major components of PUFAs (DHA and EPA), as well as clarification of the molecular mechanisms underlying their potential anti-arrhythmic roles are also required.Based on the evidence examined above, it may not be possible to propose RAS blockers, statins or PUFA (EPA and DHA) are primary AADs. However, there appears to be strong molecular plausibility and evidence base supporting their use for obtaining adjunctive anti-arrhythmic benefits alone or in combination with already established AADs.None.


Long non-coding RNAs (lncRNAs) are a diverse class of longer than 200 nucleotides RNA transcripts that have limited protein coding capacity. LncRNAs display diverse cellular functions and widely participate in both physiological and pathophysiological processes. Aberrant expressions of lncRNAs are correlated with tumor progression, providing sound rationale for their targeting as attractive anti-tumor therapeutic strategies. Emerging evidences support that lncRNAs participate in tumor-stroma crosstalk and stimulate a distinctive and suitable tumor microenvironment (TME). The TME comprises several stromal cells such as cancer stem cells (CSCs), cancer-associated endothelial cells (CAEs), cancer-associated fibroblasts (CAFs) and infiltrated immune cells, all of which are involved in the complicated crosstalk with tumor cells to affect tumor progression. In this review, we summarize the essential properties and functional roles of known lncRNAs in related to the TME to validate lncRNAs as potential biomarkers and promising anti-cancer targets. The expression of lnc-Tim3 is up-regulated in HCC patients, which is negatively correlated with the production of IFN-γ and IL-2 by tumor-infiltrating CD8+ T cells. Mechanistically, lnc-Tim3 interacts with Tim-3 to release Bat3, and thereby suppresses downstream Lck/NF-AT1/AP-1 signaling, resulting in the exhaustion of CD8+ T cells and HCC immune evasion + T by suppressing the expression of indoleamine 2,3-dioxygenase 1 (IDO1) and therefore suppresses the tumorigenesis of CRC CD8+ T cells, in response to initiate and sustain anti-tumor T cells immunity et al. et al. Dendritic cells (DCs) in the TME exhibit an important role in cross-priming CD8et al. + NKs through regulating miR-544/RUNX3, hence, enhancing the killing effects of NKs and inhibiting tumor growth. Moreover, down-regulation of GAS5 is associated with liver cancer progression, conferring a worse overall patient survival Natural killer cells (NKs) play a critical role in the anti-tumor immune response and participate in controlling tumor progression and metastasis, due to their cytotoxic potential and ability to release immunoregulatory cytokines Tumor-associated macrophages (TAMs) are key regulators of the TME, and orchestrate various aspects of tumor progression. In response to microenvironmental signals, TAMs undergo the polarization of pro-inflammatory M1 or anti-inflammatory M2 and, therefore, have anti-tumor or pro-tumor abilities Furthermore, tumor-derived CCL2 is released into the TME and recruits macrophages to promote tumor progression 6-methyladenosine (m6A)-modified regulatory network Myeloid-derived suppressor cells (MDSCs) are the central cell population with potent immunosuppressive activity on T cells Cancer stem cells (CSCs), a rare sub-population within tumor, are key components of the TME, that have the ability of self-renewal and limitless proliferation Oct4, Sox2) and the sphere-forming capacity, uncovering a role for H19 as a potential stemness regulator lncRNA H19 is one of the first non-coding RNAs identified as a cancer-related lncRNA Hox transcript antisense intergenic RNA (HOTAIR) is an oncogenic lncRNA, the expression of which is elevated in multiple CSCs The lncRNA activated in renal cell carcinoma with sunitinib resistance (ARSR) was recently identified as a novel lncRNA. ARSR is highly expressed in primary renal CSCs and predicts poor prognosis. ARSR processes self-renewal capacity and promotes the metastasis of renal CSCs. Mechanistically, ARSR interacts with Yes-associated protein (YAP) to block its phosphorylation by LATS1, thus facilitating YAP nuclear translocation, which is required to sustain CSCs self-renewal The lncRNA urothelial cancer associated 1 (UCA1) is highly expressed in multiple human cancers, including hepatocellular cancer, gastric cancer, colorectal cancer and lung cancer, which confers a worse overall patient survival Cancer-associated endothelial cells (CAEs), important components of the TME, are responsible for angiogenesis and regulation of tumor growth and metastasis + liver cancer cells package H19 inside exosomes, which is released to influence endothelial cells by promoting angiogenesis and stimulating their adhesive properties H19 expression is significantly up-regulated in glioma microvessels and glioma-induced endothelial cells (GECs). H19 regulates the proliferation, migration and tube formation of GECs by targeting miR-29a, which decreases the expression of vasohibin 2 (VASH2), an angiogenic factor MALAT1 can also regulate the properties of CAEs. Researchers have found that knockdown of MALAT1 expression significantly inhibits the proliferation of human umbilical vein endothelial cells (HUVECs), which is mediated by upregulation of miR-320a and hence, downregulation of cell cycle regulator FOMX1 expression in HUVECs HOTAIR is extremely abundant in nasopharyngeal carcinoma cells (NPC) and functions as an angiogenesis activator. Specifically, HOTAIR promotes endothelial cell tube formation and angiogenesis through directly activating the transcription of angiogenic factor VEGFA Cancer-associated fibroblasts (CAFs), the activated fibroblasts, are one of the most dominant components of the tumor microenvironment TGF-β1 secreted by CAFs induces the metastatic activity of cancer cells by regulating the expression of lncRNAs. For example, CAFs-mediated upregulation of lncRNA ZEB2NAT transcription in bladder cancer cells that promotes EMT via the secretion of TGF-β1, which is linked to poor clinical outcome In addition, CAF-promoted lncRNAs are also involved in radio-resistance and chemo-resistance. In esophageal cancer cells, CAFs-promoted expression of lncRNA DNM3OS confers significant radio-resistance via regulating DNA damage response in a FOXO1-dependent manner Here, we have summarized recent advancement involving of lncRNAs within the tumor microenvironment and their roles in the crosstalk between infiltrated immune cells, CSCs, CAEs, CAFs and tumor cells, as well as some of the underlying molecular mechanisms. lncRNAs exert their functions in different ways to modulate tumor growth and progression. As yet, only a few lncRNAs have been well-studied in the tumor-stroma crosstalk, warranting further studies on the identification of more new types of lncRNAs and their mechanisms involved in the future. A better understanding of the role of lncRNAs within the tumor microenvironment may lead to the discovery of potential biomarkers and development of novel targeted therapies.At present, the pressing issue is to systematically elucidate the key aspects of lncRNAs, including the expression, structure, function and regulatory mechanism. The improvement of analytical technologies for the specific biological functions of lncRNAs may help explore its greatest relevance to various cancers. Such information will provide a basis for considering lncRNAs as ideal diagnostic markers or even therapeutic targets. In addition, approaches of targeting lncRNAs should be considered and optimized, such as the use of siRNA to induce lncRNA degradation and CRISPR/Cas9 mediated gene editing. How to specifically deliver the respective molecules into targeted cells is still a great challenge.In summary, the functional importance of lncRNAs within tumor microenvironment is gradually characterized; clinical application of lncRNAs still needs to be studied further. With the deep-going research, lncRNAs-associated tumor-stroma crosstalk will open up a new era of anti-tumor therapy.


Hypertension is one of the major risk factors for arteriosclerosis. Anti-hypertensive peptides derived from animal proteins, such as milk, eggs and fish, are well studied. Anti-hypertensive peptides have also been identified from plant proteins such as soybeans. Rice bran, a byproduct of white rice polishing, is rich in protein and its high protein efficiency ratio is well known. This review discusses the anti-hypertensive peptides identified from rice bran protein and their mechanisms. In addition, we describe protease-digested rice bran from which functional peptides have not been isolated. Furtne (ODQ) E and itsne (ODQ) . These rThe NO-mediated vasodilation pathway is well studied and a typical pathway is shown in The origin of LRA was identified ; it was Tyr-Tyr (YY) was also identified from thermolysin-digested rice bran as an anti-hypertensive peptide ; orally ACE is in a membrane-bound form in endothelial cells, neuroepithelial cells and the brain . A solubIn addition, the renin inhibitory activity of YY was previously reported . Renin (The origin of YY was previously identified . It was 50 = 76 μM [Tyr-Ser-Lys (YSK) was identified from trypsin-digested rice bran and its ACE inhibitory activity was measured as IC = 76 μM , being sIt is well known that peptides exhibiting potent ACE inhibitory activity do not always exert strong anti-hypertensive effects . An in vAt present, no other anti-hypertensive peptides have been identified from rice bran. As described below, several reports demonstrated that protease-digested rice bran has anti-hypertensive activity, but its functional peptides are not well understood. Considering other effects, several other peptides were identified as functional peptides from rice bran. For example, VAGAEDAAK was isolated as an antioxidant peptide , LQPSHY As described above, there are few anti-hypertensive peptides identified from rice bran protein. However, the anti-hypertensive effects of protease-digested rice bran have been reported even though functional peptides were not identified. In this section, the methods for producing anti-hypertensive food material from rice bran protein, regardless of whether the functional peptides were isolated, are summarized.Bacillus thermoproteolyticus) preferentially cleaves at the N-terminal side of hydrophobic or bulky amino sidechains such as Leu, Phe, Ile and Val [Thermolysin . Anti-hypertensive effects of TRB were demonstrated in an SHR study and human clinical study. A single oral administration of TRB reduced the systolic blood pressure (SBP) of SHRs at a dose of 30 mg/kg. Repeated oral administration for 4 weeks also reduced the SBP of SHR at a dose of 50–500 mg/kg/day. The administration of TRB at 500 mg/kg/day also reduced serum troponin I levels .Administration of 1 g TRB (43 μg of LRA) for 12 weeks reduced the SBP in humans with high-normal blood pressure (SBP: 130–139 mmHg and/or diastolic blood pressure (DBP): 85–89) and grade 1 hypertension (SBP: 140–159 mmHg and/or DBP: 90–99 mmHg) . This isProtease G6, an alkaline serine endoprotease, is reported as a commercial proteolytic enzyme . The antAnti-hypertensive effects of trypsin-digested rice bran (TRP) were previously demonstrated by Wang  using a Lactobacillus helveticus and Saccharomyces cerevisiae [Aspergillus kawachii and a mixture of lactic acid bacteria  were demonstrated in an Stroke-Prone Spontaneously Hypertensive Rat (SHRSP) model and ACE inhibitory activity was observed in vivo [Fermentation is one method to produce anti-hypertensive peptides from food proteins. Anti-hypertensive peptides Ile-Pro-Pro (IPP) and Val-Pro-Pro (VPP) are cleaved from milk casein fermented by revisiae . The ant in vivo . Ferment in vivo . Intake  in vivo  and feru in vivo , are theThis was an overview of the anti-hypertensive effects of processed rice bran and the summary is presented in In conclusion, this review discussed the anti-hypertensive peptides derived from rice bran protein. LRA and YY were previously identified and predicted to originate from the same protein—vicilin-like storage protein. LRA exhibited strong vasodilating effects and promoted NO production in the endothelial layer. The mechanism underlying the anti-hypertensive effects of YY was reported as ACE inhibitory activity. Although the mechanisms of the anti-hypertensive pathway were different, these peptides demonstrated potent anti-hypertensive effects after oral administration. YSK was found from trypsin-digested rice bran and its ACE inhibitory activity was similar to that of YY. The anti-hypertensive effects and cleavage site of this peptide need to be clarified.Crude processed rice bran, which was reported to have anti-hypertensive effects, was also reviewed. This is the first review especially dedicated to the anti-hypertensive effect of rice bran digestion. TRB was demonstrated to have anti-hypertensive effects in human clinical studies and anti-hypertensive peptides were identified as a functional substance. G6RB was also reported to possess anti-hypertensive effects and to promote eNOS expression in the endothelial layer. Furthermore, TRP exerts anti-hypertensive effects. Fermented rice bran has multiple effects other than anti-hypertensive effects, such as improving glucose tolerance and adiponectin production.