Drosophila, they are instrumental for segment-specific neurogenesis and myogenesis early in development. Their robust expression in differentiated neurons implied their role in assembling region-specific neuromuscular networks. In the last decade, studies in Drosophila have unequivocally established that Hox genes go beyond their conventional functions of generating cellular diversity along the AP axis of the developing central nervous system. These roles range from establishing and maintaining the neuromuscular networks to controlling their function by regulating the motor neuron morphology and neurophysiology, thereby directly impacting the behavior. Here we summarize the limited knowledge on the role of Drosophila Hox genes in the assembly of region-specific neuromuscular networks and their effect on associated behavior.Hox genes have been known for specifying the anterior-posterior axis (AP) in bilaterian body plans. Studies in vertebrates have shown their importance in developing region-specific neural circuitry and diversifying motor neuron pools. In  Feeding, locomotion, and reproduction are some of the most fundamental behaviors exhibited by bilaterians. Regional specialization of the muscles, as well as the central nervous system (CNS) along the anterior-posterior (AP) axis, is a prerequisite for the successful and reproducible execution of these behaviors . Role ofDrosophila, Hox genes are known to establish segment-specific patterns of myogenesis and neurogenesis  which are organized into two complexes-the Antennapedia Complex (Antp-C) , proboscipedia (pb), Deformed (Dfd), Sex combs reduced (Scr), Antennapedia (Antp)], and the Bithorax Complex (BX-C) (Ultrabithorax (Ubx), abdominal-A (abd-A) and Abdominal-B (Abd-B)]. Drosophila CNS consists of the brain and segmented ventral nerve cord (VNC). Hox genes pattern the VNC, which is the Drosophila equivalent of the vertebrate hindbrain and spinal cord. The embryonic VNC specified by Hox factors can be broadly divided into five regions, namely: supra-esophageal ganglia (SPG) expressing pb and labial; sub-esophageal ganglia (SEG)  expressing Hox genes Dfd, Scr and Antp; thoracic ganglia (T1-T3 segments) expressing Antp and Ubx; abdominal ganglia (A1-A7 segments) expressing Ubx, abd-A, and Abd-B; and the terminal region (A8-A10 segments) expressing Abd-B  of VNC (Hox genes are a family of homeodomain (HD) containing transcription factors (TFs), which play an important role in determining the anterior-posterior (AP) axis of bilaterian organisms . They ar(Antp-C)  . The Ub neurons . InsteadUbx and the BX-C miRNA in self-righting (SR) motor behavior in Drosophila larvae and adults . The SRN innervated the lateral transverse (LT) muscles of the larval body wall, the LT1/2  could also be controlled by miRNA. Furthermore, it is also possible that other adult movement-associated behaviors   or Hth.Importantly, these studies show that the miRNA-dependent post-transcriptional regulation of Hox gene jumping)  may alsojumping) . Therefojumping) . Since 4behavior , therefoLastly, a tempting question is whether the miRNA/Hox genetic module could also function in MNs of other behavioral circuits like feeding, mating, courtship, grooming, and virgin/mated behavioral shift. Moreover, if such control exists, it needs to be investigated whether it is executed primarily through Hox genes or other miRNA targets.Drosophila, which generate MNs innervating the adult leg muscles. In the case of LinB lineage, the Hox gene Pb and other mTFs play an instructional role in giving unique axonal and dendritic arborization to three MNs of the lineage, thereby regulating the morphological diversity of the MNs  mutants LinA NBs, MNs were reported to undergo apoptosis. When the cell death was blocked, the surviving neurons took their fate as thoracic MNs. These MNs innervate the right target muscles and exhibit subtle arborization defects  may provide a useful mean for identifying MN specific targets genes downstream to these factors. Lastly, the distinct morphological phenotypes observed in MNs in Antp and hth mutants also suggest that Antp may be using cofactors other than canonical Hox cofactors (like Exd and Hth)  . This isand Hth) , as welland Hth) . However


On January 31, 2022, FDAapproved a Biologics License Application (BLA) for use of the Moderna COVID-19 vaccine in persons aged ≥18 years (The mRNA-1273 (Moderna) COVID-19 vaccine is a lipid nanoparticle-encapsulated,nucleoside-modified mRNA vaccine encoding the stabilized prefusion spike glycoprotein ofSARS-CoV-2, the virus that causes COVID-19. During December 2020, the vaccine wasgranted Emergency Use Authorization (EUA) by the Food and Drug Administration (FDA), andthe Advisory Committee on Immunization Practices (ACIP) issued an interim recommendationfor use among persons aged ≥18 years .Overall, the benefits for the Moderna COVID-19 vaccine outweigh any observedvaccine-associated risks  in the 7 days after vaccination .Data reviewed within the EtR Framework support the use of the Moderna COVID-19vaccine. The Work Group concluded that COVID-19 remains an important public healthproblem and that the desirable effects of disease prevention through vaccinationwith Moderna COVID-19 vaccine in persons aged ≥18 years are large andoutweigh the potential harms. With 204 million doses of Moderna COVID-19 vaccineadministered to date , but an extended interval might be considered for somepopulations. An elevated risk for myocarditis and myopericarditis among mRNACOVID-19 vaccine recipients has been observed, particularly in adolescent and youngadult males . Any person who administers or receives aCOVID-19 vaccine is encouraged to report any clinically significant adverse event,regardless of whether it is clear that a vaccine caused the adverse event. Inaddition, all COVID-19 vaccine recipients are encouraged to enroll in v-safe, a CDCvoluntary smartphone-based online tool that uses text messaging and online surveysto conduct periodic health check-ins after vaccination. CDC’s v-safe(https://www.cdc.gov/vsafe) call centerfollows up on reports to v-safe that include possible medically significant healthevents to collect additional information for completion of a VAERS report.Providers are required to report adverse events that occur after receipt of anyCOVID-19 vaccine to VAERS  granted fullapproval to the Moderna COVID-19 vaccine for persons aged ≥18years.On February 4, 2022, after a systematic review of the evidence, theAdvisory Committee on Immunization Practices issued a standardrecommendation for use of the Moderna COVID-19 vaccine in personsaged ≥18 years. CDC provided guidance that an 8-week interval betweenprimary series doses of mRNA vaccines might be optimal for some persons.Use of the FDA-approved Moderna COVID-19 vaccine is recommended for personsaged ≥18 years; benefits of the prevention of infection andassociated hospitalization or death outweigh vaccine-associated risks.


Virus infection has been consistently threatening public health. The cyclic GMP-AMP synthase (cGAS)-Stimulator of Interferon Genes (STING) pathway is a critical defender to sense various pathogens and trigger innate immunity of mammalian cells. cGAS recognizes the pathogenic DNA in the cytosol and then synthesizes 2′3′-cyclic GMP-AMP (2′3′cGAMP). As the second messenger, cGAMP activates STING and induces the following cascade to produce type I interferon (IFN-I) to protect against infections. However, viruses have evolved numerous strategies to hinder the cGAS-STING signal transduction, promoting their immune evasion. Here we outline the current status of the viral evasion mechanism underlying the regulation of the cGAS-STING pathway, focusing on how post-transcriptional modifications, viral proteins, and non-coding RNAs involve innate immunity during viral infection, attempting to inspire new targets discovery and uncover potential clinical antiviral treatments. Infectious diseases are the top threats to public health. With the persistent invading of various pathogens, mammalians have developed certain strategies to protect themselves from extraneous risks. During infection, germline-encoded pattern recognition receptors (PRRs) recognize the pathogen and damage-associated molecular patterns (PAMPs and DAMPs), such as the viral nucleic acids (DNA or RNA), which initiate subsequent immune responses, and orchestrate an elaborate defense system against infection .The cGAS-STING pathway is one of the predominant and conserved mechanisms of the host to eliminate pathogens through several aspects, including IFN-I, releasing, autophagy initiation, pro-inflammatory cytokines production, and cell death pathways . VirusesThe models of most immune evasion during innate immune responses are conserved, including altering the post-transcriptional modifications (PTMs) of vital proteins to inactivate or degrade these components, eluding DNA sensing from PRRs, decreasing cyclic GMP-AMP (cGAMP) cellular abundance, and modifying metabolism approach in host cells . IntriguAt the first stage of viral invasion, virus-derived double-stranded DNAs (dsDNAs) trigger the conformational change and activation of cGAS. Activated cGAS catalyzes and releases the second messenger 2′,3′-cGAMP, which binds to STING, an adaptor located at endoplasmic-reticulum (ER)-membrane . This prAs a signaling cascade of pathogen sensing and (IFN-I) production, IFN-β binds to IFNα/β receptor 1 (IFNAR1) and IFNAR2, which launch interferon-stimulated genes (ISGs) production through Janus kinase signal transducer and activator of transcription (JAK-STAT) pathway . ActivatAdditionally, cGAS-STING signaling participates in IRF3-independent viral-sensing and triggers autophagy, an ancestral antiviral process of host defense . The intAccumulating studies show that the cGAS-STING pathway is widely implicated in the DNA-sensing process, including viral clearance and autoimmune diseases . It is iSeveral studies showed that viruses benefit from hijacking host enzymes to alter the modification of antiviral immune cascades component proteins, thus conducive to their replication . UnderstWhen the viral DNAs are released in the cytoplasm, cGAS efficiently recognizes and processes the enthetic DNA ligands to synthesize cGAMP. Virus infection has been found to induce various forms of post-translational modifications at different sites of cGAS, which manipulates the synthetase capability of cGAS and its affinity to pathogenic DNA .To maintain the homeostasis of cGAS, several PTMs play an important role in down-regulating cGAS function to prevent its excessive activation. Upon virus infection, cGAS is phosphorylated by DNA-dependent protein kinase (DNA-PK), hindering its oligomerization and enzymatic activity . The gluin vivo  rescues cGAS inhibition by removing the small ubiquitin-like modifier (SUMO) from Lys335, 372, and 382 of cGAS . The actin vivo .Acting as the initial DNA-sensor to induce robust innate immune responses, the enzymatic activity and stability of cGAS are precisely regulated. To protect the host from the threat of virus, more PTM forms and sites of cGAS need further investigation.STING, the adaptor downstream of cGAS, is also the center molecule of most DNA-sensing pathways. Multiple PTMs have been found to dominate STING intracellular trafficking, conformational change, and activation during infection . One of STING can be ubiquitinated with K48-linked polyubiquitination, which leads to its proteasome degradation. The immune system has evolved this strategy as a negative feedback loop to balance normal immune responses and autoimmunity. TRIM29, an E3 ligase, can be induced by DNA virus stimulation, which catalyzes K48-linked ubiquitination of STING on Lys288/337 and mediates STING degradation . SimilarDistinct from K48 ubiquitination mediated degradation, K63-linked ubiquitination promotes the activation of the substrates. STING can also be modified by K63-linked ubiquitination at Lys20/150/224/236 by tripartite motif protein 32 (TRIM32), which of these are essential for STING activation and interaction with TBK1 . On the STING is standing at the crossroad of IFN-I releasing, non-classical autophagy initiation, and NF-κB activation. The ubiquitin-related regulation is quite crucial for the stability and function of STING to prevent viral immune evasion.TBK1, the downstream component of STING in the cGAS-STING axis, its kinase activity is indispensable to IFN-I generation and virus-induced autophagy initiation . Multi-cUbiquitination and acetylation are also engaged in TBK1 regulation. Virus sensing induced the expression of E3 ubiquitin ligase RNF128. K63-linked ubiquitin chains are continuously added to TBK1 sites at Lys30 and Lys401, triggering TBK1 activation and following IFN-I-β release . USP38 eSeveral enzymes engage in distinct TBK1 modifications to prevent TBK1 from excessive activation upon viral stimulation. Previous studies reveal that the Src family kinases (SFKs) Lck, Hck, and Fgr restrict IFN-I production. During virus infection, Lck/Hck/Fgr can directly phosphorylate TBK1 at Tyr354/394 to restrain TBK1 dimerization and activation as a feedback approach in antiviral immunity . RecentlIFN-β transcription upon viral infection. After translocated into the nucleus, IRF3 interacts with CREB-binding protein (CBP)/p300 to initiate downstream genes transcription  to interrupt the antiviral responses in host cells, aiming to attenuate innate immune system activation, such as the cGAS-STING axis .pUL48, a DUB encoded by human cytomegalovirus (HCMV), removes K63 ubiquitination of STING to attenuate IFN-I induction and promote carcinogenesis . TransmiThe severe acute respiratory syndrome coronavirus (Sars-CoV) membrane-anchored PLpro domain (PLpro-TM) is elucidated its function of disrupting STING-TRAF3-TBK1 complex formation, decreasing the ubiquitination level of the complex components, ultimately impairing IRF3 phosphorylation and dimerization . The papThe tegument is a protein cluster that fills the space between the nucleocapsid and the envelope expressed in all herpes viruses. The tegument proteins are essential for the viral envelope and viral DNA containing capsid transport . Apart fLately, the tegument proteins of HSV-1 have been reported to be involved in disrupting cGAS-STING induced IFN-I production. For instance, UL37 deamidates a crucial asparagine site of cGAS to eliminate cGAMP composition, deactivating IFN-I initiation cascades after infection . AnotherThe HCMV tegument protein UL23 interacts with STAT1 and hinders STAT1 phosphorylation from optimizing subsequent viral dissemination . UL94 ofAccessory proteins have different purposes and quantities in many viruses and focus more on viral spreading, evasion, and host immune regulation rather than manipulating viral replication .Upon infection, herpes simplex virus 1 (HSV-1) accessory protein Us11 prevents Hsp90 interaction with TBK1, disrupting Hsp90-TBK1 complex formation. Moreover, Us11 also induces TBK1 proteasomal degradation. These processes of Us11 facilitate HSV-1 replication by reducing IFN-I-β generation . Vpx, a Diverse viral proteins are a virus-involved strategy against host immunity. The persistent appearance of novel variants of viruses causes health emergencies to humanity. Mastering the function of viral proteins and how they manipulate the innate immune system helps us prevent viral evasion and ultimately reduce risks from virus-induced infection.RNA-centric management of host-virus interactions is increasingly causing attention . This seHost immunity engages diverse RNA-directive strategies to operate vital proteins expression in IFN-I signaling. For instance, long non-coding RNAs (lncRNAs) contain more than 200 nucleotides and regulate gene expression at transcriptional or post-transcriptional levels . After bThe miRNAs are members of small non-coding RNAs. Multiple DNA and RNA viruses are capable of encoding miRNA to accelerate their propagation or promote immune evasion in the host . CurrentRNA regulatory mechanisms that participate in virus defense immunity are potential candidates for therapeutic targeting. Numerous non-coding RNAs remain unknown. Therefore, future studies need to pay more attention to these special RNAs.There are still numerous coronaviruses that have been uncovered and need to be analyzed . In receSF, as an adjuvant for the SARS-CoV-2 vaccine. Compared with other coronaviral proteins, PLpro contributes to both virus replication and host cell signaling-cascade regulation, which is more suitable to be a target for antiviral drug design  have evolved a special vesicular trafficking method, which can translocate viral genome (vDNA) into host intranuclear environment without being detected by surveillance of abnormal DNA ligands, therefore bypassing the cGAS-STING pathway . Then, tApart from adjusting signal delivery by key proteins, several strategies regulate the intensity of antiviral response at the mRNA level. in vivo, several valuable targets in cGAS-STING signal transduction offer inspiration for antiviral drugs invention.Viruses are the mainspring of infectious diseases, several carcinogenic processes, and have caused immeasurable public health for years. Understanding how viruses adjust the innate immune system affords probabilities to cure virus-related diseases and prevent viral infection. However, simply knowing the principles and patterns is merely a beginning. Antiviral drug development is an urgent issue to humanity to diminish the impact of virus disturbance and prevent the evasion of viruses. Acting as the key checkpoints in maintaining the interferon homeostasis Recently, multiple anticancer drugs have been repurposed of their capabilities in antiviral treatments . For insNotwithstanding all these mechanisms about virus escape we have discussed above, there are still innumerable details that need further exploration. The appearance of drug-resistant variants and novel viruses with high pathogenicity are bound to immeasurable economic and public health damages. There still is a long way to go with this tug of war between humans and viruses.BioRender.com. YY, MP, QW, and YC revised the manuscript. CW edited and reviewed the final version of this manuscript. All authors contributed to the article and approved the submitted version.TH drafted the manuscript, and prepared figures with The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Hyperglycosylated cancer cells are often decorated with abundant sialic acids, which are recognized by Sialic acid binding immunoglobulin type lectins (Siglecs) expressed on immune cells. As sialic acids are normally expressed on almost all cell types, Siglecs can function as receptors for “self”. Presently, 15 human Siglecs  are known, of which most are immunoinhibitory receptors that induce strong inhibitory signaling when Siglecs bind sialic acids. Hence, tumor cells use hyper sialic acid expression in their favor to modify the immune system that can lead to immune suppression. Such engagement along the Siglec-sialic acid axis can dampen major killing modes of effector immune cells and suppress the activation of immune responses, which can lead to immune tolerance and tumor growth. In this review, we emphasized recent studies on all 15 Siglecs found in humans, many of which still remain understudied. In addition, we highlighted different strategies in disrupting the surface Siglec-sialic acid interactions, including antibodies and glycan moieties, which can steer back antitumor immune responses to reduce tumor size and improve overall survival of cancer patients.Hypersialylation is a common post-translational modification of protein and lipids found on cancer cell surfaces, which participate in cell-cell interactions and in the regulation of immune responses. Sialic acids are a family of nine-carbon α-keto acids found at the outermost ends of glycans attached to cell surfaces. Given their locations on cell surfaces, tumor cells aberrantly overexpress sialic acids, which are recognized by Siglec receptors found on immune cells to mediate broad immunomodulatory signaling. Enhanced sialylation exposed on cancer cell surfaces is exemplified as “self-associated molecular pattern” (SAMP), which tricks Siglec receptors found on leukocytes to greatly down-regulate immune responsiveness, leading to tumor growth. In this review, we focused on all 15 human Siglecs (including Siglec XII), many of which still remain understudied. We also highlighted strategies that disrupt the course of Siglec-sialic acid interactions, such as antibody-based therapies and sialic acid mimetics leading to tumor cell depletion. Herein, we introduced the central roles of Siglecs in mediating pro-tumor immunity and discussed strategies that target these receptors, which could benefit improved cancer immunotherapy. Siglecs  might hold the key to better solutions for human diseases, including cancer. Recent studies on Siglecs have shown their important roles in tumor immunosurveillance including immunosuppression, making them attractive anti-cancer molecular targets. Presently, tumor immune escape is a hallmark of tumor growth  and invoSiglecs as potential immune checkpoints are expressed on most white blood cells in the immune system  and bind+ macrophage subpopulation is found in the bone marrow, lymph node, liver, and spleen , a, a168], In contrast, Siglec-14, Siglec-15, and Siglec-16 do not have ITIM or ITIM-like motifs. These are activating Siglecs that associate with DAP12, which recruits SyK  178]. T. T178]. Tumor cells have evolved to exploit hypersialylation to trick the immune system into recognizing them as “normal self”, resulting in attenuated immune cell activity  189]. H. H189]. Despite advances in standard chemotherapy and allogenic hematopoietic stem cell transplantation , their associated non-specific toxicities and high mortality rates present obstacles for achieving durable remission in hematological malignancies. Monoclonal antibodies (mAbs) targeting Siglecs are an emerging alternative modality for passive immunotherapy to deplete tumor cells ,192. TraTraditional unlabeled  mAbs can target specific Siglec-positive cells to induce apoptosis. The expression of certain Siglec members, such as the B-cell surface-restricted Siglec-2 (CD22), has garnered growing interest for cell-directed antitumor therapies. Moreover, CD22 expression increases by 60% to 80% in B-cell malignancies , making Next, myeloid-cell-expressing inhibitory Siglec-7 and Siglec-9 play a crucial role in T-cell activation and antitumor response. Recent in vitro and in vivo studies have revealed that Siglec-7 and Siglec-9 promote tumor progression in humanized knockout murine models by supporting the immunosuppressive tumor microenvironment . MoreoveDespite improvements in the disease outcome in patients with B-cell malignancies using single-agent or tandem mAb-targeted therapy, most fail to achieve sufficient cytotoxicity and/or complete remission. Alternative therapies are needed for patients with relapse or refractory (r/r) disease (after failed standard chemotherapies or prior immunotherapies) and elderly or frail patients who are unsuitable for aggressive chemotherapeutics. To overcome these hurdles, another class of targeted therapy, tripartite antibody-drug conjugates (ADCs), was developed. ADCs harness specific mAb as a vehicle to deliver potent cytotoxic drugs to tumor cells with minimized off-target toxicity. Several Siglec-directing ADCs have been approved by the FDA for clinical use in haematological neoplasms including Inotuzumab ozogamicin  and Gemtuzumab ozogamicin , targeting CD22 and CD33 respectively C 198,19. Both AD−, Ph+) patients suffering from CD22 positive relapsed extramedullary-ALL [Motivated by the success of Mylotarg, a similar immunoconjugate targeting CD22, Inotuzumab ozogamicin, was recently approved as monotherapy for adults with r/r B-cell precursor CD22-positive acute lymphoblastic leukemia (ALL). InO’s success was partly contributed to by CD22’s ability to undergo constitutive endocytosis into B-cells following antibody ligation (for details refer to the excellent review ). InO delary-ALL . Similarlary-ALL ,207,208.lary-ALL .Immunotoxins (RITs) are recombinant proteins which express a variable fragment (Fv) of monoclonal antibody fused to either bacterial or animal toxins. Contrary to drug conjugates, immunotoxins’ cytocidal activity relies on the targeting antigen’s specificity as well as the toxin moiety’s activity to induce cell death, and with reduced dose-limiting toxicity . A CD22-+ AML are being evaluated in preclinical and early clinical trials  to determine their efficacy as immunotherapeutic agents.Bispecific antibodies (bsAbs) are a fusion of two different antibodies corresponding to two different antigens or distinct epitopes of a single antigen in various formats. One particular format targets the cytotoxic potential of immune effector cells (IECs) by bridging an activating receptor on IEC  and an antigen  on malignant cells for subsequent tumor cell lysis ,213. SupNPMI mutations were associated with high CD33 expression. Clinical trials are ongoing (NCT02520427) to evaluate the safety and tolerability of CD33/CD3 bispecific antibody in 55 patients with r/r AML. Although preliminary results from this study indicate that AMG330 showed potent anti-leukemic activity, their rapid renal drug clearance required continuous intravenous infusion (CIV) for up to several weeks [AMG330, a human BiTE (Bispecific T-cell engager) directed against CD33/CD3 led to T-cell expansion E, differal weeks . + AML cell lines [To reduce renal clearance and subsequently reduce CIV frequency, two approaches are introduced. In the first approach, AMV564, a tetravalent bispecific tandem diabodies (TandAbs) with a molecular weight (106 kDa) exceeding the renal clearance threshold showed a longer half-life than AMG330 . CD33/CDll lines . Prelimill lines . Despitell lines . + HL60 cell line and de novo AML cells independent of the E:T ratio [de novo CD33+ AML cells were enhanced in the presence of the ADAM17 inhibitor, which inhibited CD16 shedding [With a better understanding of the cytotoxic potential of NK cells in cancer therapy, tumor-specific NK cell-mediated ADCC using novel bi- and tri-specific killer engager antibodies  were generated ,220. Thr:T ratio . Additioshedding . The speshedding . The effshedding . The smaAdoptive cell transfer (ACT) is a rapidly emerging form of immunotherapy which utilizes the patient’s own immune cells  to detect and destroy malignant tumor cells. One of the ACT techniques, CAR-T cell-based treatment, is described as “a living drug" since it involves genetically modified T cells to express synthetic receptors called CARs on their surface F. CARs hBuilding on the success of CAR-T therapy in the treatment of B-cell malignancies, efforts have been made towards developing similar CAR-T strategies to improve clinical outcomes in AML patients. Despite the preclinical success of CD33-directed CAR-T-therapy in inducing anti-leukemic effects, in vivo targeting of AML blasts is difficult due to the overlapping expression of AML associated-antigens on progenitor and mature hematopoietic cells of the myeloid lineage. AEs such as hepatic damage and an immunosuppressive milieu that hindered T cell-mediated killing were also unfavorable. To circumvent these challenges, Cartellieri et al. developed the UniCAR platform, which consists of a tumor-specific inactive CAR and a targeting module (TM) that works as an on-off switch for CAR-T antitumor activity ,229. TheOver the years, specific, high-affinity sialosides-ligands have been developed to target tumor-related Siglecs as an alternative immunomodulatory strategy to halt cancer progression. Natural sialic acid-containing glycans serve as important regulators of the Siglec checkpoint in immune cell responses, involving cytoplasmic regulatory motifs (ITIM/ITAM), Siglec microdomain localization, and cis/ trans interaction via sialoglycans as ligands ,230,231.The low affinities of natural Siglec ligand binding posed a challenge in synthetic sialoside ligands for modulating Siglec function. Kelm et al. revolutionized this field by designing the first high-affinity synthetic analogues targeting CD22 derived from its natural sialic acid scaffold . SpecifiAdvances in the structural information on Siglecs further aided in the rational design of high-affinity sialic acid mimetics (SAMs). The crystal structure of Siglec-1’s  V-set domain bound to its endogenous ligand 3’sialyllactose Lac) revealed the minimal sialic acid recognition site organized by three conserved Siglec residues, W2 (strand A), R97 (strand F), and W106 (strand G) within the ligand-binding domain . AdditioOver the years, the roles of glycans in cancer have become valuable in developing antitumor therapies. SAMs are synthetic ligands which mimic the oligoantennary oligosaccharides found on endogenous glycoconjugates, and have shown potential as targeting moiety to selectively bind Siglec-expressing tumor cells . VariousSiglecs are protein receptors in glyco-immunology, which are fundamental in understanding the biology of various cancer progression in patients. Considering that hypersialylation and tumor immune evasion are in a close relationship with poorer OS in patients, more work on other understudied Siglecs, soluble Siglecs, and their tumor-associated sialic acid ligands are needed. For example, the roles of inhibitory Siglec-11 and Siglec-XII in myeloids still remain an enigma in many cancers. Additionally, new findings on the Siglec-10-CD24 axis in TAMs revealed possible sialic-acid independent pathways in the tumor microenvironment . Thus, gSiglecs are becoming widely recognized as potential targets to modulate the effector cell-signaling responses for therapeutic applications, such as in hematological malignancies ,249. Man


A total of 17 phase-II flavanone-derived metabolites were identified. No significant biomarker changes were observed. Plasma and urinary concentrations of all metabolites, including hesperetin metabolites, were higher after lemon extract intake. Total plasma metabolites showed significantly mean lower Tmax (6.0 ± 0.4 vs. 8.0 ± 0.5 h) and higher Cmax and AUC values after lemon extract intake. We provide new insights on hesperetin-eriodictyol interconversion and naringenin formation from hesperidin in humans. Our results suggest that regular consumption of a soluble and eco-friendly eriocitrin-rich lemon extract could provide a circulating concentration metabolites threshold to exert health benefits, even in the so-called low-producers.The intake of hesperidin-rich sources, mostly found in orange juice, can decrease cardiometabolic risk, potentially linked to the gut microbial phase-II hesperetin derivatives. However, the low hesperidin solubility hampers its bioavailability and microbial metabolism, yielding a high inter-individual variability (high vs. low-producers) that prevents consistent health-related evidence. Contrarily, the human metabolism of (lemon) eriocitrin is hardly known. We hypothesize that the higher solubility of (lemon) eriocitrin vs. (orange) hesperidin might yield more bioavailable metabolites than hesperidin. A randomized-crossover human pharmacokinetic study ( Epidemiological studies and meta-analyses have associated the intake of flavonoid-rich citrus fruit consumption with lower stroke risk  and the There is evidence supporting that citrus flavonoids may represent promising bioactive agents for treating metabolic and cardiovascular diseases ,11. In t® 500 mg (micronized purified flavonoid fraction of 50 mg flavonoids (expressed as hesperidin) plus 450 mg of diosmin, a semisynthetic analog) that are used in the treatment of venous insufficiency and has shown its effectivity in improving leg symptoms, edema, and quality of life in patients with the chronic venous disease . Th. Thorthool (M12) .O-glucuronyl-sulfate in humans after orange juice intake [Our study also shows the formation of eriodictyol and homoeriodictyol conjugates after hesperidin intake. Another recent study in rats highlighted the presence of these metabolites upon demethylation of hesperetin by phase-I enzymes . Other ae intake . Howevere intake , althouge intake  reportedWe have also identified naringenin metabolites  in most The high interindividual variability observed in the production of flavanone metabolites was not abolished after lemon extract intake. Although the gut microbial metabolism seemed to be favored, however, phase-I and phase-II enzymes also contribute to the high interindividual variability of polyphenols metabolism as previously suggested ,46,55. NWe acknowledge some limitations in our study. Although post-prandial vs. baseline changes were compared in each individual, we lacked a control group with no citrus extract consumption to draw unequivocal conclusions on the differences in metabolic, biochemical, and oxidative markers. Moreover, the exact comparison of the quantitative production of some flavanone metabolites was not possible since the orange extract only contained hesperidin, unlike the richer flavanone profile of the lemon extract. In this regard, it could have also been interesting to conduct a parallel study with pure compounds to evaluate how much the extract matrix influences these molecules’ pharmacokinetics. On the other hand, the intake of hesperidin allowed us to confirm some metabolic steps unequivocally. We have focused on eriocitrin and hesperidin metabolism and their phase-II flavanone metabolism. However, much more information remains to be elucidated regarding the microbial-derived ring fission phenolic catabolites upon comparing lemon extract vs. hesperidin metabolism.Taking into account the results and our limitations, we have reported new insights into the human metabolism of eriocitrin and hesperidin. Our findings support that eriocitrin rich-lemon extract intake provides more circulating phase-II flavanone metabolites and higher concentrations than after consuming hesperidin-rich extracts. Our results also suggest that lemon extract intake could provide a sufficient circulating concentration metabolites threshold to exert health benefits upon long-term consumption, even in the so-called low-producer individuals. This could be relevant to overcome the disparate flavanones-related health-effects observed in human subjects. The investigation of health-related effects after regular consumption of this aqueous-soluble and eco-friendly eriocitrin-rich extract is warranted.


Tissue donors are evaluated for communicable disease in order to minimize the risk of transmission to recipients. Although there are data suggesting SARS-CoV-2 viremia across a wide spectrum of illness, prevalence in deceased tissue donors and the potential for transplant transmission are unknown.Eight tissue banks participated in a retrospective analysis of samples from eligible deceased tissue donors from Oct 2019 through June 2020, one participant in Canada and the remainder located in the United States. All four Census regions of the continental US and all major racial-ethnic groups were represented. EDTA or sodium citrate plasma aliquots were tested in singlicate with the Research Use Only Procleix SARS-CoV-2 Assay on the Procleix Panther System, which uses transcription-mediated nucleic acid amplification (TMA) technology for detection of the SARS-CoV-2 RNA. Plasma  aliquots were sent to Grifols for an alternate SARS-CoV-2 nucleic acid amplification (NAT) test to verify reactivity and also sent for antibody testing using the emergency use authorization Ortho VITROS Immunodiagnostic Products Anti-SARS-CoV-2 Total test. The VITROS assay uses immunometric technology for qualitative measurement of total antibody  to SARS-CoV-2. The proportion of donors with confirmed RNAemia  and 95% confidence intervals were computed.Of 3,455 donor samples with valid final results, 26 (0.76%) were initially positive for SARS-CoV-2 RNA; of these, 3 were confirmed by alternate NAT. Of donor samples collected in 2019 0.00%  were confirmed RNAemic, while of those collected in 2020, 0.12%  were confirmed RNAemic. One of 26 initial positive, and none of the three samples confirmed by alternate NAT, tested positive for anti-SARS-CoV-2 Spike antibodies by serology. Infectivity studies are pending on one sample with sufficient available volume.The rate of SARS-CoV-2 RNAemia in deceased tissue donors is approximately 1 per 1,000, and it is unknown whether this RNAemia reflects the presence of infectious virus. Given these results, the risk of transmission through tissue is most likely to be low.Melissa Greenwald, MD, Alamo Biologics (Consultant)Eurofins VRL Laboratories (Consultant)Right Cell Biologics  Eduard Grebe, PhD, Gilead Sciences (Consultant)Sedia Biosciences Corporation Vitalant (Employee) Alyce Linthurst Jones, PhD, LifeNet Health (Employee) Matthew Kuehnert, MD, American Association of Tissue Banks (Board Member)ICCBBA (Board Member)Musculoskeletal Transplant Foundation (Employee)


Paraneoplastic neurologic syndromes(PNSs) caused by immune checkpoint inhibitors(ICIs) is rare and requires clinicians to differentiate between disease progression and immune-related adverse effects(irAEs). We hereby report the case of immune-related myelitis accompanied by positive paraneoplastic autoantibodies following durvalumab treatment for extensive-stage small cell lung cancer (ES-SCLC). A 70-year-old Chinese woman with ES-SCLC was administered durvalumab with etoposid-platinum(EP) as first-line treatment. Four cycles after treatment with EP plus ICI, she developed immune-related myelitis with positive paraneoplastic autoantibodies . Spinal MRI showed diffuse abnormal signal shadow in the cervicothoracic spinal cord. She was discontinued for chemotherapy, and treated with high-dose steroids, intravenous immunoglobulin and plasmapheresis, maintenance therapy with steroids resulted in a favorable neurologic outcome. This is the first report of durvalumab-related PNSs. We supposed that the development of paraneoplastic myelitis was causally related to immune activation by durvalumab. Prompt diagnosis and therapeutic intervention are essential for the effective treatment of paraneoplastic myelitis. Small cell lung cancer (SCLC) is a deadly disease that represents about 15% of all lung cancers . The staImmune checkpoint inhibitors (ICIs) have significantly increased survival and they are becoming the standard of care in many malignancies . The finMeanwhile, SCLC is most frequently associated with paraneoplastic neurologic syndromes (PNSs) . PNSs caHere, we report a patient with ES-SCLC treated with first-line durvalumab combined with etoposide-platinum for 4 cycles and developed immune-related myelitis with positive paraneoplastic autoantibodies.A 70-year-old Chinese women without any previous neurological illnesses was diagnosed with small cell lung cancer with synchronous adrenal metastases. The gene detection indicated that PD-1 was positive. She underwent four cycles of durvalumab plus etoposid-platinum chemotherapy as first-line chemotherapy. After 3 cycles therapy, computed tomography (CT) evaluation showed an almost complete tumor response Fig. . Brain MImmune checkpoint inhibitors (ICIs) is associated with a range of neurological immune-related adverse effects, including myasthenia gravis (MG), infammatory myositis, encephalitis/myelitis, meningitis, or Guillain–Barré syndrome (GBS) . Most ofRecent studies suggested that ICIs can trigger both antibody-mediated and T-cell mediated paraneoplastic neurologic syndromes . AnotherIn a series of 468 patients with and without PNS, 61 were ZIC-4 positive, including 92% of SCLC . SOX1 anFor severe neurotoxicity , current guidelines suggest management in the following order: ICI cessation, high-dose steroids, other T cell suppressive medications and intravenous immunoglobulin (IVIG) or plasmapheresis , 22. In In conclusion, we present a case with PNSs affecting the spinal cord in a ES-SCLC patient treated with durvalumab combined with etoposide-platinum chemotherapy. This is the first report of durvalumab-related PNSs. We should measure paraneoplastic autoantibodies of SCLC patients before ICIs therapy and determine whether the use of ICIs in patients with serum onconeural antibodies but without neurological symptoms in order to decrease the risk of neurological irAEs. Differentiation between disease progression and side effects may be difficult and treatment decisions should be discussed in a multidisciplinary team. Immune checkpoint inhibitors should be discontinued and treatment with corticosteroids should be initiated early as the drug of first choice. Therapy may be escalated by intravenous immunoglobulin and plasmapheresis.


Xenopus laevis eggs are a model system for studying chromosome biology. Xenopus egg extracts can be synchronised in different cell cycle stages, making them useful for studying DNA replication, DNA repair and chromosome organisation. Combining single-molecule approaches with egg extracts is an exciting development being used to reveal molecular mechanisms that are difficult to study using conventional approaches. Fluorescence-based single-molecule imaging of surface-tethered DNAs has been used to visualise labelled protein movements on stretched DNA, the dynamics of DNA–protein complexes and extract-dependent structural rearrangement of stained DNA. Force-based single-molecule techniques are an alternative approach to measure mechanics of DNA and proteins. In this essay, the details of these single-molecule techniques, and the insights into chromosome biology they provide, will be discussed.Cell-free extracts from  Xenopus laevis, have historically been used for studying vertebrate development and the cell cycle, but the number of possible manipulations was improved with the development of cell-free egg extracts . A. A22]. AVisualisation of single DNA molecules using this assay elucidated details of the eukaryotic replication reaction. Similar replication efficiencies for single- and double-tethered DNAs provided evidence that CMG helicases from the same origin do not have to remain physically associated with one another for replication . ModificThe replication of tethered DNAs in flow cells with extracts enabled development of real-time single-molecule imaging of DNA replication. A lagging-strand processing enzyme, Fen1, was fused to a photo-switchable fluorescent protein, mKikGR, and included in replication mix. Fen1-mKikGR competes with endogenous Fen1 for binding to nascent DNA and Fen1-mKikGR retention on DNA allows replication fork progression to be imaged in real-time B 25]. B. B25]. BReal-time imaging of DNA replication with Fen1-mKikGR was adapted to probe dynamics of individual nucleosomes after replication fork collision . HistoneXenopus egg extracts. Cohesin loading in extracts is dependent on licensing of DNA with Mcm2-7 DH complexes [Xenopus cohesin on to tethered λ DNAs showed that cohesin dynamically translocates after loading [The mechanism of sister chromatid cohesion establishment can be studied with omplexes . Single- loading . Cohesin loading . When grlag) following a short pause, consistent with results from ensemble experiments [lead could also be bypassed following a longer pause , possibly reflecting CMG uncoupling from the leading-strand DNA polymerase that remains stalled at the unrepaired DPClead [lead and DPClag, before, during and after collision, in the same experiment.Other encounters with DNA-bound proteins, like DNA–protein cross-links (DPCs), during replication are potentially toxic. DPCs caused by common agents  are proteriments ,32, but er pause C. In bul DPClead . In someIn new work, single molecule assays with labelled GINS have been used to investigate the signal for CMG removal during DNA replication termination . Timely Xenopus egg extracts contain components for vertebrate DNA repair pathways [pathways  and havepathways . Both LSpathways . NHEJ bepathways .To investigate the order in which protein factors synapse DSBs, single-molecule Förster resonance energy transfer (smFRET) assays were developed in the Walter and Loparo laboratories using HSS extracts . In an iA hurdle that NHEJ must overcome is ligation of non-compatible DNA ends after DSB, for instance non-complementary overhangs or ends with damaged nucleotides. Egg extracts were used to investigate the timing of end processing enzymes making DNA ends compatible for NHEJ . DNA polXenopus egg extracts are a powerful in vitro system as large volumes of cell-free extract synchronised in the cell cycle can reconstitute complex reactions involving chromosomes. Extending assays in extracts to provide single-molecule resolution gives an ability to follow discrete interactions in a single experiment [periment , measureperiment ,30 and mperiment ,15,40. Tin vivo, can be used in reconstituted eukaryotic systems, like those developed to load cohesin complexes onto DNA [Control of the cell cycle, with separation of origin licensing and firing in the HSS/NPE system, makes egg extracts useful . Althougonto DNA ,44, replonto DNA ,45–47 oronto DNA ,49. Indionto DNA . Furtheronto DNA ,33.X. laevis eggs can be used to study reactions in chromosome biology in vitro in combination with single-molecule approaches.Extracts from Surface-tethered DNAs can be replicated with the HSS/NPE system and be visualised in real-time, which has been useful for analysing the consequences of collisions between replication forks and DNA-bound proteins.smFRET assays have shown during DSB repair by NHEJ a long-range synaptic complex protects DNA ends before end-processing and ligation in a short-range synaptic complex.Extracts can be used for studying DNA organisation into chromatin and higher order structures by SMC complexes. Use of extracts to combine the study of chromatin organisation with other reactions on DNA promises to be exciting.


ABSTRACT IMPACT: Optimization of primary septal-hippocampal co-cultures facilitates studying central cholinergic synapse formation and dysfunction OBJECTIVES/GOALS: Septal cholinergic innervation to the hippocampus is critical for normal learning and memory and is severely degenerated in Alzheimer’s disease. To understand the molecular events underlying this loss, we optimized a primary septal-hippocampal co-culture system that facilitates the study of central cholinergic synapses. METHODS/STUDY POPULATION: We developed an optimized in vitro septal-hippocampal co-culture system modified from previously published protocols. Briefly, hippocampal and septal tissue were harvested from embryonic day 19 (E19) Sprague-Dawley rats, digested with 0.1% trypsin, and an equal number of cells from each region plated onto coverslips coated with poly-D-lysine and laminin at a final density of 300 cells/mm2. We use immunostaining with validated primary antibodies and a fluorescent binding assay, together with confocal microscopy, to determine the structure of cholinergic synapses that are 1) native, 2) mammalian, 3) CNS derived, 4) comprised of physiological synaptic partners, and 5) developmentally mature. RESULTS/ANTICIPATED RESULTS: After DIV21, co-cultures maintained a healthy morphology. A subpopulation of neurons strongly expressed the cholinergic markers vesicular ACh transporter (vAChT), choline acetyltransferase (ChAT), and the high-affinity choline transporter (ChT1), whereas most neurons lacked vAChT expression and were presumably glutamatergic or GABAergic. The percentage of cholinergic neurons attained in the co-culture is ˜5-7%. The size of these cholinergic neurons is strikingly similar to that reported for BFCNs in the intact brain . All sampled cholinergic neurons (28/28 neurons) also expressed molecular machinery necessary for GABA release. Staining for a cholinergic postsynaptic marker shows that 63% of the contacts made with are synaptic. DISCUSSION/SIGNIFICANCE OF FINDINGS: Primary septal-hippocampal co-cultured neurons have not been exploited extensively in the field, perhaps due to the difficulty in maintaining such cultures for extended periods. Here, we optimized an in vitro septal-hippocampal co-culture system, a powerful tool to comprehensively analyze central cholinergic synapse formation and dysfunction.


Patients with myelodysplastic syndromes (MDS) often experience chronic anemia and long-term red blood cell transfusion dependence associated with significant burden on clinical and health-related quality of life (HRQoL) outcomes. In the MEDALIST trial (NCT02631070), luspatercept significantly reduced transfusion burden in patients with lower-risk MDS who had ring sideroblasts and were refractory to, intolerant to, or ineligible for prior treatment with erythropoiesis-stimulating agents. We evaluated the effect of luspatercept on HRQoL in patients enrolled in MEDALIST using the EORTC QLQ-C30 and the QOL-E questionnaire. Change in HRQoL was assessed every 6 weeks in patients receiving luspatercept with best supportive care (+ BSC) and placebo + BSC from baseline through week 25. No clinically meaningful within-group changes and between-group differences across all domains of the EORTC QLQ-C30 and QOL-E were observed. On one item of the QOL-E MDS-specific disturbances domain, patients treated with luspatercept reported marked improvements in their daily life owing to the reduced transfusion burden, relative to placebo. Taken together with previous reports of luspatercept + BSC reducing transfusion burden in patients from baseline through week 25 in MEDALIST, these results suggest luspatercept may offer a treatment option for patients that reduces transfusion burden while providing stability in HRQoL. Myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic neoplasms characterized by ineffective hematopoiesis, progressive cytopenias, and risk of progression to acute myeloid leukemia ,2. At diTreatments for patients with lower-risk MDS are largely aimed at mitigating anemia and thereby improving their HRQoL ,10. For For patients with transfusion-dependent lower-risk MDS, a therapeutic option that can address the underlying causes of chronic anemia and reduce transfusion burden is imperative, particularly for patients who are refractory to, intolerant to, or ineligible for treatment with ESAs. Luspatercept is a first-in-class erythroid maturation agent providing clinically meaningful reduction in transfusion burden in patients with transfusion-dependent anemia due to lower-risk MDS ,19.The phase 3 MEDALIST trial (NCT02631070) compared treatment with luspatercept and best supportive care (+ BSC) to placebo + BSC in patients with transfusion-dependent anemia due to lower-risk MDS. In the first 24 weeks of the trial, transfusion independence for ≥8 weeks was observed in 38% of patients in the luspatercept + BSC arm and only 13% of patients in the placebo + BSC arm. Furthermore, a greater proportion of patients (28%) in the luspatercept + BSC arm achieved transfusion independence for ≥12 weeks compared with 8% of patients in the placebo + BSC arm . With thIn the double-blind, placebo-controlled, randomized, phase 3 MEDALIST trial (NCT02631070), patients were randomized in a 2:1 ratio to receive luspatercept (1.0–1.75 mg/kg) or placebo subcutaneously every 3 weeks for 24 weeks, plus BSC including RBC transfusions given at the investigator’s discretion . The priHRQoL data were collected prior to the administration of study drugs at each scheduled visit, independent of RBC transfusion events, using electronic tablets as the primary method and paper-and-pencil as a supplemental method. Specifically, HRQoL was assessed at screening, cycle 1 day 1 , and every other 3-week cycle B during Patients with a Revised International Prognostic Scoring System (IPSS-R) score of very low-risk, low-risk, or intermediate-risk MDS with ring sideroblasts who had been receiving regular RBC transfusions were included, as described previously . EligiblPatients were included if they received an average of ≥2 units/8 weeks of packed RBC transfusions during the 16 weeks before randomization. Patients with hemoglobin (Hb) levels ≤ 10 g/dL at the time of or within 7 days prior to administration of an RBC transfusion were included; RBC transfusions administered when Hb levels were >10 g/dL and/or RBC transfusions administered for elective surgery did not qualify as a required transfusion to meet eligibility criteria. Patients were excluded if they had a consecutive 56-day period (≥8 weeks) that was RBC transfusion-free during the 16 weeks before randomization.The intent-to-treat (ITT) population included all subjects who were randomized in the study, whereas the HRQoL-evaluable population included all subjects in the ITT population who completed the HRQoL assessments at the baseline visit  and had at least one post-baseline assessment visit.HRQoL was assessed using the European Organisation for Research and Treatment of Cancer’s Core Quality of Life Questionnaire (EORTC QLQ-C30) version 3.0  and the The QOL-E questionnaire, an MDS-specific assessment, was of exploratory interest. QOL-E domains of physical well-being, functional well-being, social and family life, sexual well-being, fatigue, and MDS-specific disturbances were included in the exploratory analyses; specific patient-reported impact of transfusion dependence and treatment side effects were reported using the MDS-specific disturbances domain. QOL-E summary scales included the treatment outcome index (TOI), which was the summary of physical well-being, functional well-being, and MDS-specific disturbances domain scores; “General”, which was the summary of all domain scores except for MDS-specific disturbances; and “All”, which was the summary of all domain scores. Scores ranged from 0 to 100. Higher scores represent better QoL across all domains and summary scales.The data-cutoff date was 1 July 2019. Descriptive statistics of the baseline HRQoL domain scores of each PRO measure and key demographic and clinical characteristics were summarized by treatment group and overall for the HRQoL-evaluable populations. Continuous variables were summarized using means and standard deviations, while categorical variables were summarized using percentages.Least-squares (LS) mean difference in change in domain scores from baseline to week 25  between luspatercept and placebo was determined using mixed-effects repeated-measures analysis. A minimal clinically important difference (MCID) within each treatment arm was defined as a ≥10-point change in score from baseline for all EORTC QLQ-C30 domains  and ≥0.5The ITT population consisted of a total of 229 patients who were randomized: 153 patients to luspatercept + BSC and 76 to placebo + BSC. The HRQoL-evaluable population, consisting of patients with baseline and at least one post-baseline EORTC QLQ-C30 assessment, was 149 patients in the luspatercept + BSC arm and 76 patients in the placebo + BSC arm. Baseline demographics and clinical characteristics of the HRQoL-evaluable population are shown in EORTC QLQ-C30 questionnaire compliance rates among ITT patients remaining on treatment were similar between luspatercept + BSC (83.6–98.0%) and placebo + BSC (79.4–100.0%) treatment groups from baseline through week 25. Baseline scores were similar between luspatercept + BSC and placebo + BSC treatment groups; the overall baseline scores for each domain are shown in The primary EORTC QLQ-C30 domains of interest of global health status/QoL, physical functioning, emotional functioning, fatigue, and dyspnea are shown in QOL-E questionnaire compliance rates among ITT patients remaining on treatment were similar between luspatercept + BSC (82.8–98.7%) and placebo + BSC (77.9–100.0%) treatment groups from baseline through week 25. Baseline scores were similar between luspatercept + BSC and placebo + BSC treatment groups; the overall baseline scores for each domain and summary scales are shown in The impact of treatment-related side effects on patients was comparable between luspatercept + BSC and placebo + BSC at week 25; 53.8% and 60.4% of patients receiving luspatercept and placebo, respectively, reported that side effects of the treatment did not disturb their daily life at all . At weekLuspatercept is a novel treatment option for patients with lower-risk MDS who have ring sideroblasts and require regular RBC transfusions . It was These findings have implications for patients with MDS who are transfusion-dependent and refractory to, intolerant to, or ineligible for treatment with ESAs. Long-term dependence on RBC transfusions may have detrimental clinical consequences, including iron overload and its associated complications of cardiac and hepatic organ failure ,15,16, wIn the present analysis, comparing luspatercept + BSC and placebo + BSC arms, no clinically meaningful differences in HRQoL through week 25 were observed within groups and between groups across all primary and exploratory domains of the EORTC QLQ-C30 and QOL-E assessments. This shows that in reducing RBC transfusions while on luspatercept, patients did not experience negative impacts on their HRQoL, contrary to what might be presumed, since transfusions provide transient relief from anemia-related symptoms. Furthermore, luspatercept significantly reduced patient-reported disturbances from RBC transfusions on daily life relative to placebo, likely owing to the reduced number of transfusions required in luspatercept-treated patients.Although the analysis was not powered to detect statistically significant differences, the maintenance of HRQoL observed in the MEDALIST trial from the present analysis is consistent with historical clinical trials which evaluated changes in HRQoL in patients with lower-risk MDS ,39,40,41There are some limitations in this analysis that should be noted. First, the majority of concepts covered by the EORTC QLQ-C30 (cancer-specific) and QOL-E (MDS-specific) instruments were not specific to the luspatercept treatment effect, particularly concepts directly capturing benefits from RBC transfusion reduction. Future research is needed in the development of specific HRQoL instruments in order to thoroughly understand the benefits of new treatments on transfusion burden in patients with MDS. Second, HRQoL endpoint data collection was set on a fixed schedule, independent of RBC transfusion events. RBC transfusions provide temporary relief and improvement in anemia-related symptoms that could positively impact HRQoL in the days following transfusion. However, these beneficial effects of RBC transfusions on HRQoL would likely have impacted the results of the placebo group, as these patients received more RBC transfusions, and the time between PRO administration and the preceding RBC transfusion was shorter. This could have further limited detection of HRQoL improvements when comparing the luspatercept and placebo arms. Third, patient Hb levels in the MEDALIST trial were maintained at a range where HRQoL changes may be insensitive to detection. It has been shown  that theLuspatercept + BSC reduced RBC transfusion burden  and tran


Bloodstream infections are associated with considerable morbidity and mortality in the United States. Most patients initially receive parenteral antibiotics for gram-negative bacteremia, and more data is emerging supporting de-escalation to oral (PO) antibiotics to complete treatment. Previous studies evaluating PO antibiotics for gram-negative bacteremia often exclude or have underrepresented immunocompromised patients. This study evaluated clinical failure in immunocompromised patients receiving intravenous (IV) antibiotics compared to patients transitioned to PO antibiotics for gram-negative bacteremia.E. coli, Klebsiella spp., Citrobacter spp., Serratia spp., Enterobacter spp., Proteus spp., P. aeruginosa. between November 4, 2017 to November 4, 2020. Patients were excluded from this study if they had polymicrobial bacteremia, no source control within the first 5 days, or an indication for prolonged duration of treatment. The primary endpoint of this study was clinical failure defined as an escalation from PO to IV antibiotics, worsening clinical status, or readmission for the same infection within 30 days of discharge. The secondary endpoints included 30-day mortality, 90-day mortality, 30-day readmission, and time to microbiologic clearance.A single center, retrospective cohort study was conducted at 446-bed academic medical center. Patients were included if they were immunocompromised and admitted with a positive blood culture for A total of 31 immunocompromised patients were included in the study with 26 patients receiving PO step-down therapy and 5 patients being continued on IV treatment for gram-negative bacteremia. There was no difference in the primary outcome of clinical failure between the PO step-down group versus the IV therapy group . The most common immunocompromised state in both groups was being HIV positive. Patients in the PO step-down group had a significantly shorter hospital length of stay .Oral step-down therapy for gram-negative bacteremia showed similar clinical failure rates to continuous IV therapy in the immunocompromised patient population and may be an option to shorten hospital length of stay.All Authors: No reported disclosures


The continuous emergence of infectious pathogens along with antimicrobial resistance creates a need for an alternative approach to treat infectious diseases. Targeting host factor(s) which are critically involved in immune signaling pathways for modulation of host immunity offers to treat a broad range of infectious diseases. Upon pathogen-associated ligands binding to the Toll-like/ IL-1R family, and other cellular receptors, followed by recruitment of intracellular signaling adaptor proteins, primarily MyD88, trigger the innate immune responses. But activation of host innate immunity strongly depends on the correct function of MyD88 which is tightly regulated. Dysregulation of MyD88 may cause an imbalance that culminates to a wide range of inflammation-associated syndromes and diseases. Furthermore, recent reports also describe that MyD88 upregulation with many viral infections is linked to decreased antiviral type I IFN response, and MyD88-deficient mice showed an increase in survivability. These reports suggest that MyD88 is also negatively involved via MyD88-independent pathways of immune signaling for antiviral type I IFN response. Because of its expanding role in controlling host immune signaling pathways, MyD88 has been recognized as a potential drug target in a broader drug discovery paradigm. Targeting BB-loop of MyD88, small molecule inhibitors were designed by structure-based approach which by blocking TIR–TIR domain homo-dimerization have shown promising therapeutic efficacy in attenuating MyD88-mediated inflammatory impact, and increased antiviral type I IFN response in experimental mouse model of diseases. In this review, we highlight the reports on MyD88-linked immune response and MyD88-targeted therapeutic approach with underlying mechanisms for controlling inflammation and antiviral type I IFN response.• Host innate immunity is activated upon PAMPs binding to PRRs followed by immune signaling through TIR domain–containing adaptor proteins mainly MyD88.• Structure-based approach led to develop small-molecule inhibitors which block TIR domain homodimerization of MyD88 and showed therapeutic efficacy in limiting severe inflammation-associated impact in mice.• Therapeutic intervention of MyD88 also showed an increase in antiviral effect with strong type I IFN signaling linked to increased phosphorylation of IRFs via MyD88–independent pathway.• MyD88 inhibitors might be potentially useful as a small-molecule therapeutics for modulation of host immunity against inflammatory diseases and antiviral therapy.• However, prior clinical use of more in-depth efforts should be focused for suitability of the approach in deploying to complex diseases including COPD and COVID-19 in limiting inflammation-associated syndrome to infection. Host sense variety of danger signals including exposure to microbial pathogens or pathogen-associated molecular patterns (PAMPs) using a family of innate immune receptors, known as pathogen recognition receptors (PRRs) such as Toll-like receptors (TLRs), the retinoic acid-inducible (RIG-I)–like receptors (RLRs)/melanoma differentiation–associated gene-5 (MDA-5), and C-type lectin receptors –5. Most −/−) were resistant to staphylococcal enterotoxin A (SEA) [MyD88 was originally known to be involved in signal transduction from TLR and IL-1R family members with exception of TLR3 , 29, 30. A (SEA)  or SEB-i A (SEA)  and show A (SEA) . In huma A (SEA) . These d1 [1 were covalently linked by a non-polar cyclohexane ring so that a six-member heterocycle ring will increase flexibility for binding to the exposed BB-loop of the TIR domain to interact with the target domain of MyD88. Compound 4210 strongly inhibited the production of pro-inflammatory cytokines in human primary cells to SEB or LPS extracted from Francisella tularensis, Escherichia coli, or Burkholderiamallei. Consistent with cytokine inhibition, in a ligand-induced cell-based reporter assay, compound 4210 inhibited B.mallei or LPS-induced MyD88-mediated NF-kB-dependent expression of reporter activity. Furthermore, results from a newly expressed MyD88 revealed that dimeric compound 4210 inhibits MyD88 dimer formation which is critical for pro-inflammatory signaling and a single administration of compound 4210 in mice showed complete protection from lethal toxin challenge. BB-loop mimetic inhibited homodimerization of MyD88 through TIR domain interaction and blocked MyD88-mediated signaling [F.tularensis [While recruitment of MyD88 is a prerequisite for inflammatory signaling and convergence point of multiple pro-inflammatory cytokines, dysregulation or overstimulation leads to inflammation-associated syndromes with severe pathological consequences to host; thus, MyD88 appears to be a unique target for therapeutic intervention of severe pro-inflammatory cytokine signaling. It has been shown that the BB-loop region acts as the mediator of the homo (adaptor-adaptor)- and hetero (receptor-adaptor)dimerization that is necessary for the function of TIR domains to induce MyD88-mediated signaling , 17, 43.1 , 46 by c1 , 48. Dimignaling . In addilarensis . It appeBesides, BB-loop mimetic using an additional alternative approach by building a protein–protein dimeric docking model of the TIR-domain of MyD88, Olson et al. identified a binding site for docking small molecules. Computational screening of 5 million drug-like compounds led to identifying a molecule T5910047 that inhibits the TIR-TIR domain interaction and attenuates pro-inflammatory cytokine production in human primary cell cultures . SubsequMario M et al. reported a synthetic peptido-mimetic compound (ST2825) modeled after the structure of a hepta-peptide in the BB-loop of the MyD88-TIR domain, which inhibited IL-1R/TLR signaling by interfering with MyD88 homodimerization of the TIR domains and did not affect homodimerization of the death domains. Oral administration of ST2825 dose-dependently inhibited IL-1beta-induced production of IL-6 in treated mice . ST2825 A spontaneous and sustained activation of MyD88-mediated via NF-κB signaling is associated with inflammation-induced cancer. Xie et al. reported that TLR/MyD88 signaling may be a therapeutic target for colitis-associated cancer (CAC) intervention and designed a MyD88 inhibitor TJ-M2010-5, which was shown to bind to the TIR domain of MyD88 to interfere with its homo-dimerization, and the TLR/MyD88 signal pathway. In a mouse model of azoxymethane/dextran sodium sulfate (AOM/DSS)—induced (CAC) in combination with TJ-M2010-5 administration, the anti-inflammation-related cancer effect of MyD88 inhibitor was investigated in vivo, and MyD88 inhibitors may be a promising therapeutic modality for treating patients with CAC .A mutation associated with nearly 1/3 of human diffuse large B cell lymphomas (DLBCLs) has been identified within MyD88 DLBCLs bearing MyD88L265P. This mutation correlates with tumor cell proliferation and survival involving spontaneous and sustained activation of NF-κB signaling. Protein–protein docking model and in silico identified MyD88 inhibitor derived small molecule compound T6167923 showed inhibition MyD88 dimer formation and were able to inhibit cell proliferation of DLBCLs bearing the MyD88L265P as measured by iso-thermal shift assay. Future studies defining the molecular mechanism of this mutation with additional human patient tumor isolates will inform and propel development of novel therapeutics to counteract both inflammation as well as tumor formation .Chronic obstructive pulmonary disease (COPD) is characterized by emphysema, small airway remodeling, pulmonary hypertension, and chronic bronchitis. Treatment with corticoids has no effects on COPD. Acute exacerbations triggered by infections play an important pathogenic role in COPD. Recent pandemic with COVID-19 disease associated with SARS-CoV-2 infections causes respiratory failure with severe lung inflammation. Thus, there is a clear unmet medical need that requires effective and safe anti-inflammatory or disease-modifying therapies for these critical conditions which are lacking. The critical role of MyD88 in pro-inflammatory signaling associated with severe inflammation especially in chronic lung diseases such as chronic obstructive pulmonary disease (COPD) is summarized in a recent review . The disViruses are highly infectious pathogens and the physiological response to virus infection is generally initiated at the cellular level following replication . After vAlthough transcriptional factors such as IRFs are the crucial transcription factors that control expression of IFN, with a viral infection, and NF-kB regulates the pro-inflammatory cytokines and type 1 IFNs during microbial infection, other cellular factors such as adaptors  and downstream kinases such as IKK-e/TBK or IKB upstream of IRFs or NF-kB are also important host factors enable transcriptional activation of IRFs and NF-kB. Viral products, such as double-stranded RNA (dsRNA), an important byproduct of replication of many viruses, are recognized by more than one PRRs such TLRs and (RLRs)/ (MDA-5), wherein activation of IRF-3 and IRF-7 is critically involved in the regulation of IFN-α/β gene induction –70. At t−/− mice were shown to have a significant higher survival rate (86%) in contrast to MyD88+/+mice (35%) after CVB3 infection or HSV-1 [−/− DCs compared to the responses in MyD88+/+ DCs following exposure to EBOV virus-like particles (eVLPs), a significant increase in IFN α/β, IRF1, and IRF7 along with increased expression of interferon-stimulated genes (ISG) observed [Host adaptor proteins play a critical role with TLR pathway of immune signaling in inducing type 1 IFN response, and particularly MyD88 is used by all TLRs, except TLR3, and is shared by IL-1 and IL-18 receptors, while TIR domain–containing adaptor-inducing IFN-β (TRIF) is solely engaged by TLR3 and TLR4. Furthermore, while recruitment and activation of the adaptor proteins MyD88 or TRIF are critical for the induction of innate immunity, activation of both MyD88 and TRIF has been demonstrated following virus infection  and ofteor HSV-1 , 80. Alsobserved . Basicalobserved . MyD88 i+ and CD8+ T cells is associated with COVID-19 [reg), and Treg cell counts in patients with COVID-19 have been shown to inversely correlate with disease severity [Since the discovery of MyD88, a considerable progress has been made on the understanding of MyD88-linked antiviral type I IFN response and other pro-inflammatory cytokine responses with many viral and bacterial infections including its spatiotemporal regulation and function. The consequence of early strong and effective innate immune responses largely leads to overall host immunity including activation of various immune cells. The balance between host immune control and viral immune evasion is pivotal to viral pathogenesis. An unbalanced immune response, characterized by a weak production of type I IFNs and an intensified release of unbalanced pro-inflammatory cytokines, contributes to the severe forms of the many viral diseases including the current COVID-19 pandemic with the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) infection , 86. TheCOVID-19  which caseverity , 94. Whiseverity . Our labseverity . Similarseverity , 97 haveThe proof-of-concept on MyD88-targeted therapeutic approach in controlling inflammatory diseases has been validated, and even first phase clinical trial against COPD was proved to be successful ; revieweIn addition, conventional antimicrobials that directly target pathogens must continue, but additional alternate approaches are also required particularly in the absence of broad-spectrum therapeutics or vaccination strategies effective for bacterial and viral infections. Development of novel host-targeted therapeutic treatment models, a complementing strategy with small molecules targeting host factors, offers a benefit for broad-spectrum host-directed therapeutic approach. Over the years, the development of such therapeutic agents targeting the host factor(s) that are critically involved in regulating the function of the immune system and/or other cellular processes are very limited. The critical barrier toward the development of such host-directed therapeutic is the identification and/or validation of appropriate therapeutic target(s) that play a critical role during a broad range of emerging/re-emerging as well as opportunistic viral/bacterial infections. In the case of targeting hyper-inflammation and imbalanced immune responses, treatment should be focused on symptomatic rather than causal, which likely reduces exacerbated tissue damage in infectious diseases. Therefore, targeting host factor critical regulating host overall immunity would likely enhance the immune response by stimulating mechanisms that are involved in host defense against the pathogen, particularly target pathways that are perturbed as a consequence of pathogen exposure and contribute to hyper-inflammation that lead to dysbalanced responses at the site of pathology . In addiAttenuation of the MyD88 signaling pathways as an anti-inflammatory strategy is clearly beneficial. In this review, our discussion focused mostly on MyD88, and its role in the regulation of host-immune signaling pathways in the context of reducing exacerbated inflammation to enhance immunomodulation and/or balance host reactions at the site of pathology likely holds promise for the selective and symptomatic treatment of infectious diseases. Based on the available published evidence, the role of immune response to SARS-CoV-2 infection indicates that the host immune response plays an important role in controlling SARS-CoV-2 infection and the immune dysregulation can significantly modify the clinical outcomes of affected patients . RecentlTaken together, the data presented in this review suggest that the pharmacological blockade with a short-term treatment with MyD88 inhibitors showed type I IFN induction while reducing acute exacerbation of various inflammatory cytokines. Given that overall imbalance in host immune system with inflammatory diseases including toxic shock, COPD as well as COVID-19 which is more to do with controlling inflammation, MyD88-targeted therapy would likely provide an advantage in treating severe inflammatory responses. At present, available data suggest the benefit of MyD88-targeted therapy with broad-spectrum antiviral type I IFN inducing properties; future in-depth efforts should be focused for the suitability of this approach in deploying to complex diseases like COPD and COVID-19 in limiting inflammation-associated syndrome to infection.


Nucleotide addition cycle (NAC) is a fundamental process utilized by nucleic acid polymerases when carrying out nucleic acid biosynthesis. An induced-fit mechanism is usually taken by these polymerases upon NTP/dNTP substrate binding, leading to active site closure and formation of a phosphodiester bond. In viral RNA-dependent RNA polymerases, the post-chemistry translocation is stringently controlled by a structurally conserved motif, resulting in asymmetric movement of the template-product duplex. This perspective focuses on viral RdRP NAC and related mechanisms that have not been structurally clarified to date. Firstly, RdRP movement along the template strand in the absence of catalytic events may be relevant to catalytic complex dissociation or proofreading. Secondly, pyrophosphate or non-cognate NTP-mediated cleavage of the product strand 3′-nucleotide can also play a role in reactivating paused or arrested catalytic complexes. Furthermore, non-cognate NTP substrates, including NTP analog inhibitors, can not only alter NAC when being misincorporated, but also impact on subsequent NACs. Complications and challenges related to these topics are also discussed. RNA viruses are a large collection of diverse, rapidly evolving viruses with a wide host range covering bacteria and eukaryotes . In rece1-S4) are usually used to help depict these microsteps (2 to S3) and translocation (S4 to S1) (Thermus aquaticus (Taq) DNA polymerase close the active site through a large-scale conformational change of their fingers domain , and translocation, while four structural reference states (S4 to S1) . In vira4 to S1) . By conts domain . S3 and reaction . Two typreaction . Similarreaction , suggestAlthough numerous NAC-state-related structures from various viral RdRPs have provided a relatively complete structural view of the cognate NTP-driven NAC, several aspects directly or indirectly related to NAC have rarely been addressed structurally in RdRPs including nucleotide addition-free translocation, intrinsic product cleavage activities, and perturbation of NAC by non-cognate NTPs and NTP analogs. As discussed below, these events are possibly related to important events including but not limited to RdRP catalytic complex dissociation, proofreading, reactivation, fidelity control, and effective intervention. In the following sections, a synoptic introduction of these events and structural challenges to approach them are discussed.1 and S4 represent the post- and pre-translocation states, respectively. In a classic NAC, a post-translocation S1 complex directs the binding of the incoming NTP, while the polymerase in the pre-translocation S4 complex needs to translocate to the next register and reopen its active site for the next NAC . In DdRPs, there is a channel to accommodate the single-stranded 3′-portion of the product RNA and transcription factors such as bacterial GreA/GreB and eukaryotic TFIIS can facilitate intrinsic endonuclease activity of DdRP to reactivate transcription . ProofreExcept for recently reported backtracked SARS-CoV-2 RdRP structures obtained using RNA scaffolds with designed mismatches at the 3′-portion of the product RNA , structui) is the byproduct of the NTP/dNTP-driven phosphoryl transfer reaction in NAC. Under certain circumstances , PPi can reverse the reaction through pyrophosphorolysis  reverse transcriptase (RT), a non-cognate NTP can induce the cleavage of the product strand 3′-nucleotide, forming a dinucleotide with a 5–5′ poly-phosphate (tetra- or tri-phosphate) linkage when the incoming cognate dNTP is not available . Similari- and NTP-mediated cleavage activities by RdRP itself may play key roles in maintaining viral genome stability and keeping the virus away from the error-catastrophe threshold , a high-fidelity DNA polymerase  at enzymology and structural levels is a key not only to identify repurposed NA drugs, but also to design new NA drugs (The successful usage  of nucleNA drugs . As an eNA drugs . A delayNA drugs , demonstNA drugs . The nonNA drugs . For exaNA drugs . Unlike To date, structures of RdRP-RNA complexes containing non-cognate NTP-derived mismatches are rarely reported, while related structures depicting intervention mechanisms of representative NAs have been emerging with the fast growing of RNA virus research . NAs witRepresentative NAC-related RdRP-RNA complex structures have been reported in picornaviruses, HCV, influenza viruses, bacteriophage ϕ6, and more recently in bunyaviruses and CoVs . However


A progressive series of chronic microhemorrhages and inflammatory insults on the cerebrovascular system and neural-network integrity can lead to significant neurological and cognitive impairments following mild-to-moderate traumatic brain injury (mTBI). Evidence-based effective treatment is needed to stop this progression. Seventeen studies in this Research Topic  have investigated the mechanisms, diagnosis, and novel therapies of TBI-induced neuropathology and cognitive impairments. Activated inflammatory microglia, elevated Inflammatory cytokines , oxidative stress, blood-brain barrier (BBB) disintegration, microvascular inflammatory injury-induced de-coupling between high metabolic demands and compromised cerebral blood flow (CBF)/perfusion, and brain network senescence/dysfunction associated with hyper-phosphorylated tau tangling appear to be the important pathological mechanisms following TBI. These disruptions of normal brain function underlie the disorders of consciousness, insomnia, depression, and neurological and cognitive impairments that follow mTBI. These pathological changes can now be determined by innovative assays and neuroimaging technologies. The advances in this field are leading to innovative treatments for the disorders which follow mTBI.Developing approaches include intraneuroendoscopic removal of subdural hematomas, protecting BBB integrity by methylene blue or by medicinal herbs, controlling neuroinflammation by endocannabinoids, improving CBF/hypoperfusion by photobiomodulation or by executive function training (including computerized training and playing mahjong) or by transcranial direct current stimulation (tDCS), improving insomnia by cognitive-behavioral therapy. All of these approaches appear to be promising interventions to improve post-traumatic symptoms and cognitive deficits associated with mTBI.As long-term repetitive sub-concussive impacts induce subtle neuromotor dysfunction in animals, similarly, clinicopathological changes and dementia in a 93-year-old street boxer are likely due to the brain concussions that occurred many years before. Accordingly, early diagnosis and treatment could mitigate the damage from repeated sub-concussive impact-induced chronic brain inflammation and resulting subjective cognitive decline (SCD) that had occurred in early life. Measures are developing which will likely provide effective prophylactic approaches that prevent dementia.Si et al. reviewed recent progress in SCD research and its relevance to TBI-induced cognitive deficits. SCD is characterized by early self-perceptive cognitive decline before the detection by objective tests. If identified, SCD can be treated/reversed more easily than later stages of progression from mild cognitive impairment (MCI) to dementia.SCD, now recognized as an occasional preclinical sign of Alzheimer's disease (AD) and dementia, may be common in people with a history of TBI. Sun et al. showed that serum CCL2 level was elevated over 3 months and was associated with the severity of post-concussion symptoms in 52 patients with acute mTBI compared to 54 healthy subjects, whereas elevated IL-1β was associated with worsened working memory in the acute phase of TBI. Thus, elevated inflammatory CCL2 and IL-1β could be predictors of post-concussion symptoms and cognitive outcomes of mTBI.Shang et al. showed that IL-1β effectively induced cellular senescence in rat astrocytes that was accompanied by increased total and phosphorylated tau. Similarly, both oligomerized amyloid β (Aβ) and H2O2 induced cellular senescence in rat astrocytes by NLRP3-mediated IL-1β secretion. They proposed that adverse stimuli-induced NLRP3 activation and IL-1β production are potentially diagnostic biomarkers and therapeutic targets for AD/dementia.By studying inflammatory factors, Shang et al. reported reduced serum miR-451 levels in patients with intracerebral hemorrhage (ICH) compared to healthy controls. They further evaluated the role of miR-144/451 cluster in ICH mice model. Deletion of miR-144/451 cluster exacerbated neurological deficits and brain edema, significantly promoted TNF-α and IL-1β secretion and oxidative stress in the perihematomal region of knockout mice compared with wild-type ICH animals, supporting a neuroprotective role of miR 144/451against ICH-induced inflammation and oxidative stress.Yang et al. presented clinicopathological evidence of the coexistence of AD with hyper-phosphorylated tau-pathology, chronic traumatic encephalopathy (CTE), dementia with Lewy bodies, and hippocampal sclerosis with TDP-43 pathology in a 93-year-old former street boxer with a pre-mortem diagnosis of severe dementia. CTE is a tauopathy in which multifocal perivascular phosphorylated tau aggregates accumulate in astrocytes, neurons, and neurites at the depths of the cortical sulci. These results suggest that early repetitive sub-concussive impacts may trigger pre-mortem pathologies of CTE, AD, and dementia.Lavender et al. report that highly repetitive sub-concussive impact with a lightweight drop (25 g) onto anesthetized female rats for 12 weeks induced detectable/subtle neuromotor dysfunction i.e., increased foot slips in beam-walk and rotarod tests. Thus, long-term repetitive sub-concussive impacts may also be a mechanism underlying clinicopathological changes of the 93-year-old street boxer reported by Yang et al..Tanaka et al. reviewed how modulation of the endocannabinoids (eCBs) system  could facilitate the homeostatic microglia to adopt either the (M1) state, which secretes mediators of the proinflammatory response, or to the (M2) state, which mediates the resolution of neuroinflammation and tissue repair/remodeling in neuropathological conditions. They propose that cannabinoid 2 receptor signaling pathway plays a critical role in shifting the microglia from the pro-neuroinflammatory (M1) state to the anti-inflammatory/neuroprotective (M2) state.Du et al. showed in a non-randomized controlled trial that the intraneuroendoscopic technique (INET) is significantly better than traditional burr hole drainage (BHD) in the treatment of subacute and chronic septal subdural hematoma in terms of hematoma recurrence rate and subdural drainage tube (SDT) placement duration at 6-months post-operative follow-up.Wang et al. proposed MRI as the first choice for suspected VCI to evaluate brain atrophy, infarction, white matter hyperintensity, and hemorrhage. Resting-state functional magnetic resonance imaging (rs-fMRI) can detect spontaneous brain functional activity, probe the pathogenesis of VCI in-depth and provide a reference for early diagnosis and prognostic evaluation.Chronic microvascular injury/microbleeding following TBI may lead to inflammation and vascular cognitive impairment (VCI) which is difficult to diagnose. In their review paper, Shen et al. demonstrated that methylene blue (MB) treatment can reverse mitochondrial dysfunction following oxygen glucose deprivation/reoxygenation (OGD) in PC12 cells, reduce neuronal apoptosis and improve blood-brain barrier (BBB) integrity in a fluid percussion TBI mouse model. MB also inhibited ROS production, stabilized neuronal mitochondrial membrane potential (MMP), increased ATP production, and preserved brain functions following TBI as demonstrated by Morris water maze, rotarod, and modified Neurological Severity Score (mNSS) tests.Quinn et al. applied pseudo-continuous arterial spin labeling (pCASL) magnetic resonance imaging techniques in measuring CBF in 24 subjects with mild and moderate TBI before and after 10 days of computerized executive function training (CEFT) combined with active or sham anodal transcranial direct current stimulation (tDCS). Robust improvements in depression, anxiety, attention, and executive function were observed in both active and sham tDCS groups. tDCS stimulation was associated with static/increased CBF in the right inferior frontal gyrus whereas sham tDCS reduced CBF. Neuropsychological performance and behavioral symptoms, however, were not associated with changes in CBF, probably due to the lack of a control for CEFT. Thus, cerebral perfusion measured with MRI presents a potential pathophysiological target for rehabilitation paradigms of TBI.Reduced cerebral blood flow (CBF) and perfusion may underlie persistent post-traumatic symptoms of mild-moderate TBI, but these parameters might be difficult to measure. Pape et al. showed in a randomized controlled trial that familiar auditory sensory training  facilitated changes in neural connectivity in patients with TBI-induced disorder of consciousness. They highlighted a relation between language-related neural networks and improvement in awareness.Zhang et al. showed that playing mahjong for 12 weeks improved the executive function of elderly (74.3 ± 4.3) with mild cognitive impairment (MCI). Since mahjong is a popular 4-people game, further study should determine if mahjong can improve cognitive deficits in people with mild TBI.In a randomized control trial and by using MoCA, STT, and Functional Activities Questionnaire (FAQ), Dietch and Furst provide evidence supporting effectiveness of cognitive-behavioral therapy for insomnia (CBT-I)  and propose more CBT-I studies in patients with mTBI because insomnia is a highly treatable symptom of mTBI that could have broad positive impacts on the recovery of TBI symptoms including cognitive deficits.Up to 50% of TBI patients suffer from insomnia which may underlie chronic inflammation and is unlikely to subside even after symptoms of mTBI remit. Chao et al. reported that 8 weeks of PBM intervention [810-nm light-emitting diodes at 10 or 40 Hz by an intranasal and four transcranial modules targeting nodes of the default mode network (DMN)] increased brain volumes and cerebral perfusion, improved functional connectivity and neuropsychological test scores in a professional hockey player with a history of concussions and chronic symptoms of headaches, mild anxiety, and difficulty concentrating.Photobiomodulation (PBM) is a therapy that uses red-to-near-infrared (NIR) light to heal injured tissue. Lee et al. showed that herbal medicine (HM), more commonly used in China, Asian, and other developing countries as either monotherapy or as adjunctive therapy, are significantly better than conventional treatment (CT) on post-concussion syndrome , activities of daily living, neurological dysfunction, and safety profile. However, no differences in traumatic brain edema, posttraumatic hydrocephalus or cognitive dysfunction are found. As most HM studies reported were low-quality due to heterogeneity of the clinical populations, in diagnosis and in the use of active herbal ingredients for TBI treatment, further rigorous standardized RCTs of HB are needed.By meta-analysis of 36 randomized controlled trials, Adamson et al. in collaboration with Enhancing Neuro Imaging Genetics Through Meta-Analysis (ENIGMA) have emphasized the importance of international collaboration and domestic capacity building for innovative research, training clinicians and researchers and databases development to bring developing countries into the international platform for dementia and TBI research.As most TBI studies are done in wealthy countries with few data from developing countries, As we learn more about the evolving neuropathological processes and mechanisms underlying the post-traumatic syndromes following mTBI, better targeted interventions are more likely to be developed. The innovative work published in this Research Topic of Advances in Diagnosis and Treatment of TBI-Induced Neurodegeneration and Cognitive Deficits suggest that evidence-based anti-inflammatory, anti-hypoperfusion, and anti-anxiety/insomnia therapies show promise for TBI-induced post-traumatic symptoms and cognitive deficits.All authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.Biotech.com LLC. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.GX was employed by company Lotus All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. 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