HIV-infected and HIV-exposed uninfected (HEU) children have an increased risk of measles that may be due to altered immune responses or suboptimal timing of measles vaccination. We aimed to evaluate the safety and immunogenicity of measles vaccination in HIV-infected and HEU children.For this systematic review and meta-analysis, we searched PubMed, Embase, Cochrane Library, CINAHL, Global Health Library and IndMED on May 9, 2018. Studies were included if they reported on safety or seroresponse (either seroprotection/seropositivity/seroconversion) after measles vaccination in HIV-infected or HEU children. We calculated pooled estimates to compare immunogenicity outcomes between HIV-infected, HEU and HIV-unexposed children, using risk ratios [RRs] (with 95%CIs). PROSPERO registration number: CRD42017057411.I2 = 85.9%) and HEU children , which was mitigated by antiretroviral therapy and time interval between vaccination and serology. HEU and HIV-unexposed children had similar seroresponses. Vaccination at 6-months resulted in similar proportions of HIV-infected children having seroresponse compared with HIV-unexposed  and HEU children .Seventy-one studies met the inclusion criteria . Twenty-eight studies reported on safety; vaccine-associated adverse events and deaths were uncommon. Sixty-two studies reported on immunogenicity, 27 were included in the meta-analysis. HIV-infected children had lower seroresponse rates after primary vaccination compared with HIV-unexposed (RR 0.74; 95%CI: 0.61–0.90, Primary measles vaccination at 6-months of age may provide protection against measles during early infancy in settings with high prevalence of maternal HIV-infection, however, further studies are needed to evaluate this strategy in HEU children and HIV-infected children receiving antiretroviral therapy.South African Research Chairs Initiative of the Department of Science and Technology and National Research Foundation in Vaccine Preventable Diseases; Medical Research Council: Respiratory and Meningeal Pathogens Research Unit. Despite measles being targeted for elimination, outbreaks of measles continue to occur in low-middle income and high income countries. Contributing to this is under-immunization of children, as well as a shift in measles epidemiology towards infection of infants < 9 months of age, who are not generally targeted for measles vaccination. Young infants may be at increased risk of infection due to changes in maternal immunity, which nowadays is predominantly derived from vaccination rather than natural infection, thereby reducing transplacental transfer of protective antibodies from mother to fetus and lowering protection during early infancy. This might be further exacerbated in settings with a high prevalence of maternal HIV-infection, where there is waning of maternal immunity in HIV-infected women, that also results in lower concentrations of measles antibodies being transferred to their fetuses. Hence, HIV-exposed infants, including those who are HIV-exposed uninfected (HEU), are at increased susceptibility to measles infection during early infancy. This calls for a review of measles immunization strategy, particularly in settings with high prevalence of maternal HIV-infection, to inform future deliberations on alternate measles vaccine dosing schedule strategies.One previous systematic review and meta-analysis on the safety and immunogenicity of measles vaccination in HIV-infected children included studies up to February 2009. Since then, antiretroviral treatment (ART) has become widely available in many countries and the number of HEU children has increased globally due to effective Prevention of Mother-to-Child Transmission programs.ClinicalTrials.gov was accessed for ongoing and unpublished trials. The inclusion criteria were limited to observational or interventional studies in HIV-infected or HEU children that measured safety or antibody seroresponses after measles vaccination. For inclusion in the meta-analysis a comparison group was required. Case reports were included for assessment of safety.We did a systematic review and meta-analysis on the safety and immunogenicity of measles vaccination in HIV-infected and HEU children. We searched seven databases  for articles in English, French, German, Spanish, Portuguese, or Dutch published before 9 May 2018, using the key words (“measles” and “vaccine”) and “HIV”. Reference lists of the articles that were included in full-text screening were searched manually to identify additional studies. The online database The meta-analysis showed that HIV-infected children were less likely to serorespond after primary measles vaccination compared to HIV-unexposed or HEU children, while HEU and HIV-unexposed children had similar immune responses. When vaccinated at 6-months of age, similar proportions of HIV-infected and HEU children had a seroresponse compared to HIV-unexposed children. We found that vaccine-associated adverse events and deaths were uncommon.Our study builds on the previous systematic review by incorporating additional evidence published since 2009 (nine new studies on safety and 15 new studies with comparison group on immunogenicity). To our knowledge, this is the first meta-analysis on this topic which compares measles immunogenicity outcomes considering both age at vaccination and number of doses received. We further extended previous work through detailed subgroup analyses to explore heterogeneity in seroresponse estimate and improve the robustness of the evidence by using GRADE to assess quality of evidence. HIV-infected children had a reduced immune response to primary vaccination in absence of ART, when measuring immunogenicity as seroprotection and if serology was assessed more than 3 or 6-months post-immunisation.In order to sufficiently protect children born to HIV-infected mothers, primary vaccination at 6-months of age is recommended. Our findings are in line with World Health Organization recommendations to administer the primary dose of measles vaccine at 6-months of age in areas with high incidence of HIV-infection and measles, followed by two routine doses according to the national immunization schedules. However, we only identified three studies evaluating measles vaccination at 6-months of age in HIV-infected and HEU children, underlining the need for further investigation before widely adopting an early vaccination strategy. Future studies should evaluate immune responses to early measles vaccination and long-term waning of immunity in HEU children and HIV-infected children treated with ART in settings with high incidence of measles and HIV.Alt-text: Unlabelled Box1In 2015, an estimated 1.4 million births occurred in HIV-infected women, of which more than 95% lived in low- and middle-income countries (LMICs) HIV-infected children have an increased risk of severe measles disease and complications compared with HIV-unexposed children A previous systematic review and meta-analysis on the safety and immunogenicity of measles vaccination in HIV-infected children undertaken by Scott et al. included studies up to February 2009 This systematic review evaluated the safety and immunogenicity of measles vaccine in HIV-infected and HEU children, and compared immunogenicity outcomes taking age at vaccination and number of doses received into consideration.22.1This systematic review and meta-analysis adhered to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines ClinicalTrials.gov.We searched PubMed, Embase, Cochrane Library, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Global Health Library , and IndMED on 9 May 2018, for articles containing (“measles” and “vaccine”) and “HIV” (Supplementary data 1). Additional studies were identified by searching reference lists of the articles included in full-text screening and Studies were eligible for inclusion in the systematic review if they reported on immunogenicity or safety of any measles vaccination strategy in HIV-infected or HEU children aged 0–18 years. For inclusion in the immunogenicity meta-analysis, studies needed to report on primary or booster vaccination and had to include a comparator group of either HIV-uninfected children (HEU/HIV-unexposed) or HIV-infected children on a different antiretroviral therapy (ART) regimen. No restrictions regarding geographical region or year of publication were applied. Eligible study designs were interventional or observational. For assessment of safety, case reports were also included. Animal studies, systematic reviews, narrative reviews, reports of proceedings and publications not written in English, French, German, Spanish, Portuguese or Dutch were excluded.The outcomes of interest were immunogenicity and safety. Immunogenicity: studies were included if data were reported as proportions of subjects with seroprotective (≥ 330 mIU/mL or as indicated by authors), seropositive, or seroconversion (4-fold rise in titre or change from seronegative to seropositive) measles antibody responses. A composite outcome for seroresponse was created using seroprotection rates post-vaccination, and if not available, seropositivity or seroconversion rates were considered. Safety: all reported safety outcomes post-vaccination were considered, including deaths, severe adverse events (SAEs) other than death and adverse events (AEs).Two independent reviewers  screened titles and abstracts of identified studies. Articles were retained if they met the inclusion criteria according to one or both of the reviewers. In case of duplicate publications of the same results, the most complete reference was included.2.2Data were extracted from manuscripts using a standardised data extraction form (Supplementary data 2) and authors were contacted in case of missing data. Data of interest included: study design, study population, vaccine type, age at vaccination, time-period between vaccination and measurement of the serological response, number of vaccine doses administered, use of ART, outcome measures, laboratory methods used to detect measles antibodies, serological cut-off values, proportions with seroresponse, and number and type of (S)AEs.The Cochrane Risk of Bias Tool was adapted to enable evaluation of observational studies (Supplementary data 3) When multiple time-points were reported for immune responses after the same vaccine dose, the time-point closest to vaccination was reported, except for two studies that had a smaller sample size at the earlier time-point I2 > 50%), a random-effects model was applied. To explore statistical variation and heterogeneity between trials, pre-specified subgroup analyses were performed based on outcome (seroprotection), serological test, use of ART, study design, age at vaccination and time interval between vaccination and measurement of the serological response. Meta-regression was used to explore between-study variance not explained by the covariates and risk of publication bias was assessed using normal and contour-enhanced funnel plots if ten or more articles were included in the meta-analysis. Small study effects were evaluated using Egger's-test for asymmetry.Three different primary meta-analyses compared serological responses in HIV-infected vs. HIV-unexposed, HIV-infected vs. HEU and HEU vs. HIV-unexposed children using risk ratios (RRs) and 95%CIs stratified by vaccination dose and age at vaccination. In case of significant heterogeneity  system for rating overall quality of evidence 2.3The funder of the study had no role in study design, data collection, data analysis, data interpretation, or writing of the report. The corresponding author had full access to all the data in the study and all authors had final responsibility for the decision to submit for publication.3We identified 897 unique articles . SeventyIncluded study designs were randomised controlled trial (RCT) (n = 1) Taking all studies together, 15,363 children vaccinated against measles were evaluated, of which 4867 were HIV-infected, 2733 were HEU, and 7763 were HIV-unexposed.Thirty-five studies with comparison groups reported post-vaccination seroresponses in HIV-infected children, of which twelve administered ART (Supplementary data 5.1). HIV-infected children showed similar seroresponse rates after primary vaccination at 6-months  compared with later time points: 9-months , 12-months  and > 12-months of age . The pooled point estimate of HIV-infected children with seroresponse after booster vaccination was similar when administered at ≤ 24 months  or > 24 months . Two studies assessed the effect of different ART-regimens on the response to primary vaccination HEU children receiving primary vaccination at 12-months  or > 12-months of age  tended to have better seroresponse compared with HEU children vaccinated at 6  or 9-months  of age (Supplementary data 5.2).Similar to HEU children, a trend towards improved seroresponse was observed in HIV-unexposed children receiving primary vaccination at > 12-months  compared with 6-months  or 9-months of age  (Supplementary data 5.3).I2 = 85.9%)  A. SeroreI2 = 89.6%)  B, irrespI2 = 77.1%) (Twenty-one studies compared immunogenicity after primary measles vaccination between HIV-infected and HEU children. Nine studies reported significant RR estimates < 1 = 77.1%) A. The prHIV-infected and HEU children showed similar immune responses after booster measles vaccination  B [35], , [Twenty-eight studies reported on safety . In totaTwenty-three studies provided information on post-vaccination SAEs other than death in HIV-infected children (period of observation ranged 1–4 weeks post-vaccination). SAEs other than death were reported in 29 of 884 HIV-infected children (3.3%), 2 of 1337 HEU (0.1%), and 18 of 1898 HIV-unexposed children (0.9%). None of the verifiable SAEs were vaccine-related. One study reported a possible, but unverifiable vaccine-related SAE Of the 71 studies, 59 (83%) had unclear or high-risk of confounding bias and 55 (77%) had unclear or high-risk of attrition bias due to incomplete outcome data. The origin of data and the clarity of outcome definition had low-risk of bias in 60 (85%) and 54 (76%) studies, respectively (The funnel plot for comparisons containing ten or more studies (HIV-infected vs. HEU children after primary vaccination) had an asymmetrical appearance (Supplementary data 15a). The contour-enhanced funnel plot showed that studies were missing in regions of both low and high statistical significance (Supplementary data 15b), suggesting that the asymmetry cannot be explained by publication bias. Smaller studies were likely to have contributed to funnel plot asymmetry (Egger's test p = 0.009).4This review assessed the safety and immunogenicity of measles vaccination in 4867 HIV-infected, 2733 HEU and 7763 HIV-unexposed children. HIV-infected children had 26% (95%CI 10%–39%) lower seroresponse rate to primary measles vaccination compared with HIV-unexposed children, and 22% (95%CI 12%–31%) lower rate compared with HEU children. Differences between groups were no longer present after booster vaccination Primary measles vaccination with standard titre measles vaccine at 6-months of age resulted in similar seroresponse rates between groups of HIV-infected Pooled RRs showed no difference between HIV-infected and HIV-unexposed or HEU children after primary vaccination when limiting the meta-analysis to studies that administered ART, reported on seroprotection, or measured serology within 3 or 6-months post-vaccination. Thus, reduced seroresponse to primary vaccination may particularly be evident in HIV-infected children when using a less stringent serological cut-off (seroconversion or seropositivity instead of seroprotection), in the absence of ART, or after a longer time-period between vaccination and serology.Studies with different timing for ART initiation showed improved immune responses to booster vaccination in HIV-infected children after ART initiation HIV-exposed children showed a non-significant trend towards improved serological response when vaccinated at 6-months of age compared with HIV-unexposed children. This could be explained by reduced transplacental transfer of antibodies from HIV-infected women, resulting in lower levels of maternal antibodies in the infant and less interference with the B-cell response to vaccination HIV-infected children experienced slightly more SAEs other than death in the first 4-weeks post-vaccination compared with HEU or HIV-unexposed children. However, due to absence of direct comparisons between vaccinated and unvaccinated HIV-infected children and poor quality of reporting, limited conclusions can be drawn from this analysis. HIV-infected children may experience more SAEs due to their underlying illness, unrelated to vaccine administration.A previous systematic review and meta-analysis of 39 studies analysing safety and immunogenicity of measles vaccination in HIV-infected children searched literature up to February 2009 Strengths of this review and meta-analysis are the comprehensive search in seven databases and the large number of studies identified. Also, this is the first meta-analysis on this topic to separately analyse primary and booster dose by age at vaccination.Our results need to be interpreted in the context of the risk of bias evaluation and low to very low quality of evidence. All studies included in this review were of observational nature, except for one RCT In the different meta-analyses, substantial heterogeneity between studies was detected. Therefore, pooled results should be viewed as an average representing a wide distribution of seroresponses. Differences in the definition and cut-off points for serological outcomes partly explained the large heterogeneity. Due to inconsistent outcome reporting across studies, we used seroresponse, a composite of seroprotection, seropositivity or seroconversion. We encourage consistency in reporting to allow for comparison between studies.The findings from this review support the 2017 recommendations by the World Health Organization to administer an initial dose of measles vaccination at 6-months of age in areas with high incidence of HIV-infection and measles, followed by two routine doses EM, MCN, KKG participated in the conception, design and implementation of the study. EM and MvR performed screening and data extraction. EM did the statistical analysis. EM wrote the first draft of the report with input from MCN, MvR, KKG, DEG and SAM. All authors have approved the final manuscript.MCN reports personal fees from Pfizer and non-financial support from Sanofi outside the submitted work. SAM reports grants from Medical Research Council South Africa, grants from Department Science and Technology/National Research Foundation during the conduct of the study; grants and personal fees from the Bill and Melinda Gates Foundation, grants from GSK, grants and personal fees from Sanofi, grants from Pfizer outside the submitted work. All other authors declare no competing interests.


The discovery of highly active antiretroviral therapy (HAART) in 1996 has significantly reduced the global mortality and morbidity caused by the acquired immunodeficiency syndrome (AIDS). However, the therapeutic strategy of HAART that targets multiple viral proteins may render off-target toxicity and more importantly results in drug-resistant escape mutants. These have been the main challenges for HAART and refinement of this therapeutic strategy is urgently needed. Antibody-mediated treatments are emerging therapeutic modalities for various diseases. Most therapeutic antibodies have been approved by Food and Drug Administration (FDA) mainly for targeting cancers. Previous studies have also demonstrated the promising effect of therapeutic antibodies against HIV-1, but there are several limitations in this therapy, particularly when the viral targets are intracellular proteins. The conventional antibodies do not cross the cell membrane, hence, the pathogenic intracellular proteins cannot be targeted with this classical therapeutic approach. Over the years, the advancement of antibody engineering has permitted the therapeutic antibodies to comprehensively target both extra- and intra-cellular proteins in various infections and diseases. This review aims to update on the current progress in the development of antibody-based treatment against intracellular targets in HIV-1 infection. We also attempt to highlight the challenges and limitations in the development of antibody-based therapeutic modalities against HIV-1. The development of HAART has significantly reduced AIDS-related death cases . This stAntibody-mediated therapy has emerged as a promising therapeutic strategy against various infectious diseases and cancers to counteract the abovementioned limitations. In the past 15 years, considerable attention has been drawn to the development of monoclonal antibodies (mAbs) against cancers ,10. SimiThis section describes the important functions of viral proteins essential for viral replication and the accessory proteins that play roles in virus dissemination, which make them ideal targets for the development of various antiviral treatments. The therapeutic antibodies targeting these intracellular proteins will be discussed individually in HIV-1 Gag precursor (Pr55Gag) is composed of N-terminal matrix (MA/p17), capsid CA/p24), nucleocapsid (NC/p7), C-terminal p6, and spacer peptides Sp1 and Sp2 . Followi, nucleocHIV-1 Nef is a viral accessory protein. Nef proteins are not directly involved in the viral replication, but they play essential roles in viral pathogenesis at both early and late phases. The functions of Nef in disease progression has been extensively reviewed ,48,49,50HIV-1 Tat is known to play a pivotal role in HIV-1 replication and infectivity by mainly activating the transcription from the viral long terminal repeat (LTR) promoter by binding to the transactivation-responsive region (TAR) hairpin in the viral RNA transcript . Das andenv gene [b]pyridine-2-carboxamide and 4-amino-6-methoxy-2-(trifluoromethyl)-3-quinolinecarbonitrile (termed as 103833 and 104366) . F. F128]. Moreover, other arising issues of application of small-fragment antibodies as an antiviral therapy include the production cost and potential cytotoxicity . The higThe rise of HIV-1 resistance towards HAART is a global issue . CurrentAlthough HAART remains the standard therapeutic intervention to control HIV-1 infection, the treatment is unable to clear the virus and requires life-long administration which may eventually lead to drug resistance . AntibodIn addition, current advancement in antibody engineering also allows the development of single antibody molecules that can target more than one viral antigen known as bi-, tri-, multi-specific antibodies, these may contribute to future antibody therapy compared to the use of multiple single-target antibodies ,151,152.While HIV-1 infection is still prevalent in developing countries, the advancement of technology plays an important role in addressing these problems to bring affordable therapy into primary care in the most remote settings. Other limitations of antibody-mediated therapies include off-target toxicities, poor bioavailability and stability, high renal clearance rate, and high individual-to-individual variation as mentioned above. Continuous efforts in discovering novel and robust viral targets must be continued to counter-act newly emerging viral resistance. HIV-specific cell-penetrating antibodies are more effective than small molecule inhibitors. These therapeutic modalities can simultaneously target both extra- and intra-cellular targets, and efficiently block and neutralize HIV-1 activities, hence, resulting in a more comprehensive antiviral action. As highlighted in this review, several challenges remain to be addressed and advanced technologies are anticipated to kick in to improve the current antiviral strategies. Continuous efforts are needed in seeking novel antibody-based therapies against HIV-1 and combination therapy is one of the promising approaches based on the current findings to control the emerging resistant HIV-1 variants and to prolong the feasibility of antibody-mediated HIV treatment.


Generating a knowledge base of new activity cliffs (ACs) defined on the basis of compound set-dependent potency distributions, also taking confirmed inactive compounds into account.Different AC definitions, representations and search criteria were rationalized and applied.For nearly 100 different target proteins, for which medicinal chemistry and biological screening data were available, target set-dependent ACs were identified. More than 20,000 target set-dependent ACs and associated information are made freely available.As more compound data become available for new targets, the search for target set-dependent ACs, including confirmed inactive compounds will continue. Second-generation ACs will be subjected to systematic structure–activity relationship analysis. In medicinal chemistry, activity cliffs can be rationalized just like geographical landscape features. Highly potent compounds occupy the top of mountains in activity landscapes and weakly potent compounds the surrounding valley floors. A small chemical change may be sufficient to push a highly potent compound off a steep cliff and transform it into a weakly potent or inactive valley floor compound. Hence, by studying activity cliffs, one can often understand which chemical modifications have large-magnitude effects on biological activity. We report a systematic search for a new type of activity cliffs using compound data from medicinal chemistry and biological screening. Shown are two activity cliffs involving the same highly potent compound (top). One cliff is formed with a weakly potent structural analog  and the other with an inactive analog . The analogs are active and confirmed inactive, respectively, against matrix metalloproteinase 14. In medicinal chemistry, activity cliffs (ACs) are defined as pairs or groups of structurally similar compounds  having large differences in potency . AccordiFor the assessment of substructure-based similarity, that is, the presence or absence of well-defined substructures in compounds that are compared, the concept of matched molecular pairs (MMPs) ,5 is par50 or (assay-independent) equilibrium constants . For AC analysis, equilibrium constants are strongly preferred to avoid the use of assay-dependent readouts when determining potency differences. An at least 100-fold difference in potency (two orders of magnitude) has often been applied as a general threshold for AC formation MMP-cliffs in the following) were defined by applying a constant similarity and variable target set-dependent potency difference criterion according to :Similariset-dependent) ACs (sd(R)MMP-cliffs). By definition, sd(R)MMP-cliffs were exclusively formed by active compounds. Only target sets with statistically significant potency variations according to  ACs) and corresponding isd(R)MMP-cliffs involving inactive analogs are introduced.Results of a systematic search for these target set-dependent ACs are summarized.The relevance of sd(R)MMP-cliffs and smaller numbers of isd(R)MMP-cliffs for structure–activity relationship exploration is emphasized.An open access deposition containing >20,000 target set-dependent ACs is described.Different frequencies of sd(R)MMP- and isd(R)MMP-cliffs are rationalized.The number of sd(R)MMP- and isd(R)MMP-cliffs is expected to increase.Second-generation ACs will be subjected to systematic structure–activity relationship analysis.


More than 200,000 Veterans transition between hospital and skilled nursing facility (SNF) annually. Capturing outcomes of these transitions has been challenging because older adult Veterans receive care at VA and non-VA hospitals, and four different kinds of SNFs: VA-owned and -operated Community Living Centers (CLCs), VA-contracted community nursing homes (CNHs), State Veterans Homes (SVHs), and non-VA community SNFs. We used a novel data source which concatenates VA, Medicare, and Medicaid data into longitudinal episodes of care for Veterans, to calculate the rate of adverse outcomes associated with the transition from hospital to SNF in all enrolled Veterans age 65 and older undergoing this transition 2012-2014. The composite primary outcome included Emergency Department (ED) visits, rehospitalizations, and mortality (not in the context of hospice) within 7 days of hospital discharge to SNF. We used multivariable logistic regression to adjust for Veteran and hospital characteristics and hospital random effects. In the 388,339 Veterans discharged from 1502 hospitals in our sample, we found more than 4 in 5 Veteran transitions (81.7%) occurred entirely outside the VA system. The overall 7-day outcome rate was 10.7%. After adjustment, VA hospitals had lower adverse outcome rates than non-VA hospitals . VA hospital-CLC transitions had the lowest adverse outcome rates; in comparison, non-VA hospital-CNH  and non-VA hospital-CLC  had the highest rates. These findings raise important questions about the VA’s role as a major provider and payer of post-acute care in SNF.


Optical imaging plays an indispensable role in biology and medicine attributing to its noninvasiveness, high spatiotemporal resolution, and high sensitivity. However, as a conventional optical imaging modality, fluorescence imaging confronts issues of shallow imaging depth due to the need for real-time light excitation which produces tissue autofluorescence. By contrast, self-luminescence imaging eliminates the concurrent light excitation, permitting deeper imaging depth and higher signal-to-background ratio (SBR), which has attracted growing attention. Herein, this review summarizes the progress on the development of near-infrared (NIR) emitting self-luminescence agents in deep-tissue optical imaging with highlighting the design principles including molecular- and nano-engineering approaches. Finally, it discusses current challenges and guidelines to develop more effective self-illuminating agents for biomedical diagnosis and treatment. Imaging techniques such as single-photon emission computed tomography (SPECT), positron emission computed tomography (PET), magnetic resonance imaging (MRI), computed tomography (CT), optical imaging, ultrasound imaging, and photoacoustic (PA) imaging have become powerful tools to detect and monitor the physiological or pathological processes at the molecular, subcellular, cellular, tissue and body levels  to generate high-energy peroxides such as 1,2-dioxetanone, 1,2-dioxetanedione, and 1,2-dioxetane firstly. Then the peroxides dissociate directly or modulating by an acceptor molecule (F) through energy transfer, leading to the production of an excited state (an excited carbonyl or acceptor molecule) and subsequent light emission.Self-luminescence imaging, which does not rely on real-time light excitation and thus eliminates the tissue autofluorescence and photo-bleaching, has attracted increasing attention in recent years. By virtue of the merits, self-luminescence imaging displays higher imaging sensitivity, higher SBR, and deeper imaging depth relative to fluorescence imaging . To dateNear-infrared (NIR) self-luminescence imaging has attracted increasing enthusiasm due to higher tissue penetration depth of NIR light than visible light, which dramatically expands the visualization scope of physiological or pathological processes in living subjects in a noninvasive way , Renilla luciferase (Rluc), and Nanoluc (Nluc)  to achieve higher imaging depth due to the decreased light scattering and tissue absorption of NIR photons through living tissues relative to that of visible light , 2) by inserting a carbon-carbon double bond between the thiazole group and benzothiazole group of luciferin (LH2). The substrate iLH2 produced light in the NIR range (λmax = 670 nm) in presence of a native Fluc   in the presence of luciferase  that were matched with a mutant luciferase (CBR2), generating NIR bioluminescence (730 nm for NH2-NpLH2 and 743 nm for OH-NpLH2)  system enabled stable and highly resolved NIR bioluminescence imaging of the migration of cells in the brain  served as an energy donor and quantum dots (QDs) acted as an energy acceptor  were developed for in vivo NIR bioluminescence imaging by combining BRET and fluorescence resonance energy transfer (FRET)  is an alternative approach to red-shift the light emission from visible into the NIR region. To achieve this, a NIR-emitting dye or nanoparticle was commonly selected to covalently conjugate with the natural bioluminescence system as an acceptor to allow for efficient energy transfer  by integrating CRET and FRET for detecting peroxynitrite (ONOO−) and hydrogen peroxide (H2O2) -alt-4,7-bis(thiophen-2-yl)benzo-2,1,3-thiadiazole)  acted as both the CRET acceptor and the FRET donor. A cyanine dye  was embedded into the nanoparticles to serve as the FRET acceptor, inducing the NIR chemiluminescence. SPN-CF was anchored with galactose-attached copolymer  favoring hepatocytes-targeting  of 5 and 10 nM, respectively , which performs a great deal of versatility and flexibility in experimental systems  as an energy acceptor was doped into SPN-PFPV to obtain SPN-PFPV-NIR with 778 nm emission by CRET. The SPN-PFPV-NIR allowed the ultrasensitive detection of H2O2in vivo in both peritonitis and neuroinflammation models  to pair with the substrate bis oxalate (TCPO) . in vivo FA imaging in mice, which confirmed that this probe could detect clearly endogenous FA released from the folate metabolism  with dicyanomethylene-4H-chromene (DCMC) as an electron acceptor was synthesized  possesses less light-tissue interactions  than those in NIR-I window, permitting imaging with higher SBR and sensitivity   was generated by the light irradiation of PPVs and then oxidized the vinylene bond to form an unstable PPV-dioxetane intermediate, which could spontaneously decompose into PPV-aldehyde fragments along with photonic efflux  that could be unfavorable for efficient accumulation in tumor. To solve it, Pu et al. prepared an amphiphilic PPV polymer with grafted poly(ethylene glycol) that could self-assemble into smaller nanoparticles (24 nm) in aqueous solution  (Jiang et al., in vivo detection of biological and pathological processes in living systems. With the development of new self-luminescence agents, the selectivity and sensitivity of bioluminescence, chemiluminescence and afterglow imaging tools have been dramatically improved.Self-luminescence including bioluminescence, chemiluminescence and afterglow have permitted imaging with higher sensitivity and deeper penetration depth than that obtained by fluorescence because it avoids the requirement of real-time light excitation and eliminates tissue autofluorescence. As NIR light possesses improved penetration depth than that obtained by visible light, many strategies via molecular- and nano-engineering approaches have been adopted to develop NIR self-luminescence. Stimulated by the merits, NIR self-luminescence imaging has been applied for Because bioluminescence, chemiluminescence and afterglow agents for deep-tissue optical imaging in living system is in the infancy, some challenges still need to be addressed. Firstly, the luminescence intensity of NIR self-luminescent agents is relatively low due to the limited efficiency to red-shift luminescence emission from visible to NIR window. Thus, it is necessary to explore new agents with evolving molecular- or nano-engineering strategy to obtain brighter luminescence. Secondly, compared with NIR-I light, NIR-II light has substantially lower light scattering, and thus increases the imaging sensitivity and tissue penetration depth (Jiang and Pu, QM and MG conceived this manuscript. QL, QM, JZ, and MG wrote and edited this manuscript.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.


Cerebral amyloid angiopathy-related inflammation (CAA-ri) is a rare variant of CAA with autoimmune inflammation. A 77-year-old female experienced light-headedness during walking and mild ataxic gait without any other objective neuropsychological deficits. Brain magnetic resonance imaging (MRI) revealed an area of abnormal signal and mild parenchymal swelling in the right parietal lobe, indicating vasogenic edema. T2⁎-weighted gradient echo imaging revealed some subcortical microbleeds in the same lesion. Based on the proposed criteria for CAA-ri, she was diagnosed with probable CAA-ri. After 4 months, the spontaneous improvement was noted in the patient's clinical and radiological findings. This report presents a rare and atypical case of CAA-ri in which the diagnosis was established after the patient underwent neuroimaging for only mild neurological symptoms, and the patient's clinical and radiological findings displayed spontaneous improvement. Despite typical and striking MRI findings of CAA-ri, this patient only presented a minimal symptom; this dissociation could highlight the significance of not misinterpreting any new neurological symptoms. Thus, increased availability of MRI and growing awareness of CAA-ri might result in more incidentally diagnosed cases in the future. Furthermore, this case suggests that it would be better to strictly monitor the clinical-radiological findings of patients with probable CAA-ri who only present with minimal symptoms without the initiation of immunosuppressive therapy. Clinical symptoms of CAA-ri include subacute mental status changes (59%), headaches (35%), focal deficits/seizures (24%), dementia (12%), hallucinations (12%), and stroke-like signs [β autoantibodies are reportedly elevated during the acute phase of CAA-ri, which is considered to result from an inflammatory response to Aβ peptides in the cerebrovascular walls [Cerebral amyloid angiopathy (CAA) is an aging-related disease often reported in the elderly population; in this disease, sel wall . CAA causeizures 4%, demeneadaches 5%, focal∗-weighted gradient echo (T2∗-GRE) imaging revealed some subcortical microbleeds in the right parietal lobe, and postgadolinium T1-weighted images exhibited no enhancement. Considering her clinical characteristics and MRI findings and excluding the differential diagnosis as infection and brain tumor, she was diagnosed as probable CAA-ri based on the proposed criteria for CAA-ri [A 77-year-old female with hypertension and hyperlipidemia experienced staggering while walking 3 months ago. She fell and bruised her head 2 months ago. At that time, head computed tomography revealed hypodensity in the right parietal region. She presented with a feeling of light-headedness during walking to our hospital. Neurological examination revealed only mild ataxic gait without any other objective neuropsychological deficits. In addition, no cognitive impairment was recognized. Brain MRI revealed an area of abnormal signal and mild parenchymal swelling in the bilateral and asymmetric parietal lobe. Precisely, subcortical hyperintensity was noted in the bilateral and asymmetric parietal lobe on T2-weighted and fluid-attenuated inversion recovery (FLAIR) images and apparent diffusion coefficient (ADC); however, these lesions were not recognized in diffusion-weighted imaging (DWI), indicating vasogenic edema . Moreover CAA-ri . We neitHere, we present the case of a patient with CAA-ri who was diagnosed after undergoing neuroimaging for mild neurological symptoms; however, this diagnosis was probable based on the proposed criteria . Moreoveβ-amyloid deposits of precursor protein categorize CAA into sporadic and inherited types, with the sporadic Aβ-type CAA accounting for the majority of cases. Although several symptoms are asymptomatic, CAA causes cerebral hemorrhage, cerebral infarction, and transient neurological symptoms, such as abnormal sensation and weakness. Reportedly, sporadic Aβ-type CAA is not a rare disease as its prevalence increases with aging with approximately 30% of autopsy cases of asymptomatic elderly patients exhibiting the sporadic feature [CAA implies amyloidosis in the small- and medium-sized blood vessels in the meninges and brain.  feature .Even more rarely, CAA causes CAA-ri, an inflammatory reaction that is clinically characterized by acute to subacute progressive cognitive dysfunction, headache, positive visual symptoms, coma, seizures, and focal neurological deficits that may be severe . There i∗-GRE imaging, diagnosis of CAA-ri is facilitated through frequent occurrence of microbleeds below the cortex and subcortex [∗-GRE imaging revealed some punctate low-signal lesions in the cerebral subcortex, and microhemorrhages have remained after the resolution of inflammatory reaction, as previously reported [Diagnosis of CAA-ri without invasive brain biopsy is facilitated by MRI, which would typically reveal asymmetrical swelling of the white matter and T2 hyperintense abnormalities, both of which adequately mimic brain tumors, necessitating their clinical differentiation. On T2ubcortex . In our reported . In centreported . As ADC β immunotherapy for AD and represents vasogenic edema and/or sulcal effusion on FLAIR sequences, as well as hemorrhagic findings [β autoantibodies play a pivotal role in the etiopathogenesis of ARIA, and CAA-ri is broadly accepted as a human spontaneous model of the therapeutic-induced ARIA [Remarkably, this case presents a dissociation between mild symptoms and notable and typical brain MRI findings of CAA-ri, indicating the significance of not misinterpreting any new neurological symptoms in patients who have already experienced an episode of CAA-ri to receive timely diagnosis and therapy, resulting in better clinical outcome , 13. Hadfindings , 15. Repced ARIA , 10. Howced ARIA . In our ced ARIA . Perhapsced ARIA , 19. Thuβ peptides resulting in the formation of Aβ autoantibodies in the cerebrovascular walls, the underlying cause of this autoimmune response remains unclear [β autoantibodies from the vasculature to the brain parenchyma. Antibody binding to vascular Aβ peptides triggers inflammation by trapping Aβ during efflux from the brain. Consequently, it becomes increasingly vulnerable to blood–brain barrier (BBB) damage, microhemorrhage, and vasogenic edema [Remarkably, both clinical and radiological findings demonstrated spontaneous improvement in our patient. Typically, patients with CAA-ri are highly responsive to immunosuppressive therapies, including corticosteroids, or others such as methotrexate, mycophenolate mofetil, cyclophosphamide, and intravenous immunoglobulin if promptly diagnosed and medicated –22. Alth unclear . Howeveric edema , 24. Theic edema . Likewisic edema , it is iThis report describes a rare and atypical case of CAA-ri, in which patient was diagnosed after undergoing neuroimaging because of only mild neurological symptoms, and the patient's clinical and radiological findings showed spontaneous improvement without immunosuppressive therapy. Despite typical and striking MRI findings of CAA-ri, this patient only presented a minimal symptom; this dissociation between mild symptoms and remarkable and typical brain MRI findings of CAA-ri highlights the significance of not misinterpreting any new neurological symptoms. Thus, increased availability of MRI and growing awareness of CAA-ri might result in more incidentally diagnosed cases in the future. Furthermore, this case suggests that it would be better to strictly monitor the clinical-radiological findings in patients with probable CAA-ri who only present with minimal symptoms without the initiation of immunosuppressive therapy.


Laser-Stimulated Fluorescence (LSF) is used to identify fully fledged feathering in the hatchling enantiornithine bird specimen MPCM-LH-26189, supporting precocial nesting behavior in this extinct group. The LSF results include the detection of a long pennaceous wing feather as well as cover feathers around the body. The LSF technique showed improved detection limits over and above synchrotron and UV imaging which had both been performed on this specimen. The findings underscore the value of using a wide range of analytical techniques. This previous study found no feathers or chemical evidence for plumage  with faint ribbing visible in a yellowish stain suggested to be more consistent with the morphology of vegetal material than with feathers (see Supplementary Note 1 of1). MPCM-LH-26189 is reasonably well articulated and has some soft-tissue-associated chemistry1. These lines of evidence were used to suggest that MPCM-LH-26189 might have been largely featherless when it died . While unconfirmed  this implies a developmental strategy towards the altricial end of the altricial-precocial developmental spectrum, which would be unexpected evidence of enantiornithine reproductive behavior. Given the exceptional preservation of the Las Hoyas fossil record2, and of feathers in particular4, here we evaluate the developmental strategy of MPCM-LH-26189 using Laser-Stimulated-Fluorescence (LSF)5 to identify possible additional evidence of preserved feathers. For instance, in MPCM-LH-26189 yellowish stains are preserved in various positions across the entire body , but their structural details were not apparent using those techniques. Inspection of the matrix . This suggests the possibility that there is too low a percentage of carbon to be easily visible in the matrix, but under LSF, the residual carbon (or possibly the detected iron1) quenches the fluorescence25 in the local area revealing the feather filaments.Synchrotron generated x-ray beams were applied to MPCM-LH-261895, has helped to clarify the developmental strategy of MPCM-LH-26189 and of enantiornithines more generally. This example underscores the range of data that remains undiscovered in important fossils, and the value of adopting a broader analytical repertoire.In bridging detection limits in synchrotron and UV analytical techniques, Laser-Stimulated Fluorescence (LSF), a rapid, low-cost techniqueet al.5, which was developed in29. Thus, only an abbreviated version is provided here. A 405 nm laser diode was used to fluoresce the specimen follow standard laser safety protocol. 30 second time-exposed images were taken with a Nikon D810 DSLR camera and a 425 nm laser blocking filter. Post processing applied uniformly across entire images  was performed in Photoshop CS6 software.Laser-Stimulated Fluorescence (LSF) imaging followed the protocol of Kaye


This report describes a case of elderly patient with right iliac artery fissured pseudoaneurysm, leading to inferior vena cava arteriovenous fistula that was treated by covered stent device deployed at the level of the right iliac pathologic segment. A 77-year-old male patient underwent open repair aorto-bi-iliac graft for abdominal aortic aneurysm (AAA) 7 years ago. On clinical follow-up, an abdominal bruit was observed. The patient underwent contrast medium-enhanced multidetector computed tomography (MD-CT) that revealed in the arterial phase, at the level of the right iliac anastomosis, the presence of fissured pseudoaneurysm (arrowhead inIliac artery–inferior vena cava or iliac vein AVF, due to iliac anastomotic pseudoaneurysm, is a rare clinical complication after AAA open repair.


Cancer immunotherapies that remove checkpoint restraints on adaptive immunity are gaining clinical momentum. Approaches aimed at intrinsic cellular immunity in the tumor microenvironment are less understood, but are of intense interest, based on their ability to induce tumor cell apoptosis while orchestrating innate and adaptive immune responses against tumor antigens. The intrinsic immune response is initiated by ancient, highly conserved intracellular proteins that detect viral infection. For example, the RIG-I-like receptors (RLRs), a family of related RNA helicases, detect viral oligonucleotide patterns of certain RNA viruses. RLR activation induces immunogenic cell death of virally infected cells, accompanied by increased inflammatory cytokine production, antigen presentation, and antigen-directed immunity against virus antigens. Approaches aimed at non-infectious RIG-I activation in cancers are being tested as a treatment option, with the goal of inducing immunogenic tumor cell death, stimulating production of pro-inflammatory cytokines, enhancing tumor neoantigen presentation, and potently increasing cytotoxic activity of tumor infiltrating lymphocytes. These studies are finding success in several pre-clinical models, and are entering early phases of clinical trial. Here, we review pre-clinical studies of RLR agonists, including the successes and challenges currently faced RLR agonists on the path to clinical translation. Regs), tumor associated macrophages (TAMs) and myeloid derived suppressor cells (MDSCs)   , are expDDX58) . When inDDX58) –26 and aDDX58) –29. A haDDX58) , leadingDDX58) . TogetheDDX58) . InteresDDX58) –35. In t] Figure . SynthetRIG-I was first identified as a cytosolic DExD/H box RNA helicase activated in response to certain RNA viruses . RIG-I iOnce engaged, MAVS signaling activates three kinases that serve as regulators of inflammation, Inhibitor of κB-Kinase (IKK)-γ, TANK-Binding Kinase (TBK)-1 and IKK-ε –58. These.g. polyinosinic-polycytidylic acid, or poly(I:C)], a ligand that also activated some Toll-Like Receptors (TLRs). In contrast to RIG-I and MDA-5, the other RLR known as Laboratory of Genetics and Physiology (LGP)-2 lacks the CARD domain shared by RIG-I and MDA-5, but is otherwise similar to the other RLRs  , 80 was BCL2. Delivery of this construct to cells potently stimulated IFN production and NK activation, while enhancing tumor cell killing through Bcl-2 ablation [MYC) that are currently ‘undruggable.’Exciting innovations within the field of RIG-I agonists are emerging. For example, a powerful, bimodal application of RNAi-based silencing of intra-tumoral gene targets using a 5′-triphosphate modified dsRNA sequence would allow for RIG-I activation and simultaneous gene targeting. This approach was demonstrated in melanomas, using 5′-3p-siRNA sequences specific to the anti-apoptotic gene ablation . This coablation . A similablation . DefininDespite the potential success of RIG-I and RLR agonists, the immune system is powerful and incompletely understood, warranting cautious optimism and thorough examination of the caveats associated with innate immune activation, including possible on-target induction of autoimmunity, or induction of a cytokine ‘storm’ which could pose a threat to patient safety –94. It iin vivo have been reported only recently. For example, a study employing a minimal 5′-triphophosphorylated stem-loop RNA (SLR) sequence delivered by intra-venous delivery to mice activated in RIG-I signaling, IFN induction, and expression of genes required for potent anti-viral immunity in vivo. A recently described ‘conditional’ RIG-I ligand, in which the 5′-triphosphorylated terminus of the RNA duplex remained shielded until release by predetermined molecular cues in vivo, could enhance delivery of RIG-I agonist to tumors, and minimize RIG-I activation outside of the TME [Delivery of small nucleotide sequences to tumor cells and leukocytes within the TME is another major obstacle to the widespread utility of RIG-I or RLR-based therapeutics in the cancer setting. Studies aimed at generating stable, specific and potent RIG-I ligands that retain functionality  the TME . However the TME .Therapeutic RIG-I and RLR agonists are emerging as a novel approach to engage the immune system in the fight against cancer. Importantly, RIG-I signaling directly promotes tumor cell killing through three distinct modes of action: intrinsic apoptosis, extrinsic apoptosis, and pyroptosis. Further, simultaneous activation of the innate and adaptive arms of the immune system may generate durable therapeutic responses. The multi-faceted mechanisms by which RLR agonists eliminate cancer cells represent the well-rounded arsenal of weapons required to fight aggressive and metastatic cancers effectively.


Adoptive T-cell immunotherapies, including chimeric antigen receptor-modified T-cells (CAR-T cells), have revolutionized cancer treatment, especially for hematologic malignancies. Clinical success of CAR-T cell monotherapy in solid tumors however, has been only modest. Oncolytic viruses provide direct cancer cell lysis, stimulate systemic immune responses, and have the capacity to provide therapeutic transgenes. Oncolytic virotherapy has shown great promise in many preclinical solid tumor models and the first oncolytic virus has been approved by the FDA for the treatment of advanced melanoma. As monotherapies for solid tumors, oncolytic virotherapy provides only moderate anti-tumor effects. However, due to their complementary modes of action, oncolytic virus and T-cell therapies can be combined to overcome the inherent limitations of each agent. This review focuses on the aspects of oncolytic viruses that enable them to synergize with adoptive T-cell immunotherapies to enhance anti-tumor effects for solid tumors. Clinical use of adoptive cell therapies to treat cancer has gained great interest in recent years, adding new treatment options to the paradigm of surgery, radiotherapy, and chemotherapy. Bolstering a patient's immune system with infused T-cells that have been genetically modified to specifically target tumor cells holds great promise and has demonstrated clinical efficacy in hematologic malignancies . These TAdoptive cell transfer of autologous CAR-T cells targeting B-cell antigen CD19 have resulted in profound remission in patients with refractory B-cell malignancies. Recently, the first chimeric antigen receptor CAR-T cells, Tisagenlecleucel , have beOncolytic viruses (OVs) have been designed to selectively replicate in and kill cancer cells. It is well established that OVs can stimulate adaptive immune responses to tumor cells due to the release of tumor associated antigens (TAAs), pathogen-associated molecular patterns (PAMPS), and danger-associated molecular patters (DAMPS) from lysed tumor cells. These responses also shift tumors from cold (immune desert) to hot (inflamed) tumors . Once pr+ T cell infiltration and resulted in 50% survival within 30 days, compared to treatment with heat-inactivated oVSV or mice left untreated whose median survival was approximately 20 days. Similarly, infusion of OT-I (OVA-specific) T-cells resulted in 50% survival within 30 days. To enhance the antitumor effect, the authors combined oVSV treatment with systemic infusion of OT-I T-cells resulting in a more potent anti-tumor response than either single agent treatment, approximately 70% survival at 50 days  . In heteBispecific T cell engagers (BiTEs) are molecules consisting of a CD3-scFv linked to another scFv specific for an antigen expressed on the surface of tumor cells. By utilizing these molecules, tumor resident/infiltrated T-cells can be redirected toward additional specific antigens expressed on cancer cells. Blinatumomab is an FDA approved CD19 BiTE for the treatment of relapsed or refractory B-ALL  which fuTo increase the efficacy of BiTE molecules and decrease unwanted side effects due to constant systemic administration, local constitutive expression of BiTEs at the tumor site would provide stimulation for tumor resident T-cells without systemic toxicity. To this end, OVs have been used to express various BiTE molecules, providing a retargeting moiety to T-cells together with virus mediated oncolysis. To target tumor cells expressing the EphA2 antigen, an oncolytic vaccinia virus (VV) was engineered to express an EphA2 BiTE, called T-cell engager armed VV (TEA-VV). In an orthotopic lung tumor xenograft model, when human PBMCs were delivered together with the EphA2.TEA-VV, tumor growth was significantly reduced compared to mice receiving only oncolytic VV or unarmed oncolytic VV with PBMCs .ex vivo activated, adoptively transferred T-cell accumulation and proliferation in a subcutaneous model of colorectal carcinoma. Administration of unarmed Onc.Ad provided oncolysis and reduced tumor growth which was significantly enhanced by the addition of the BiTE molecule in the presence of activated T-cells  expressing an EGFR-BiTE (Onc.Ad-EGFR.BiTE), derived from cetuximab which is used clinically to treat colorectal  and head T-cells . However T-cells . Treatmereatment   induces high expression of multiple pro-inflammatory chemokines  resulting in increased accumulation and persistence of adoptively transferred OT-I T-cells in both immune competent and incompetent models .The suppressive tumor microenvironment is depleted of pro-T cell cytokines which is a significant inhibitory mechanism tumors develop to evade cytotoxic T-cells. OVs can deliver molecules to stimulate T-cells at the tumor site and reverse this anergy. Administration of an Onc.Ad expressing TNFa and IL-2 (Onc.Ad-IL2/TNFa) in five consecutive doses has significant antitumor effect in an immune competent Syrian hamster model of pancreatic cancer . SubsequTo increase the efficiency of CAR-T cell trafficking and persistence within tumors, production of proinflammatory chemokines and cytokines from the tumor mass has been investigated. An Onc.Ad expressing both IL-15 and RANTES (Onc.Ad-IL15/RANTES) has demonstrated that combining both molecules can have a profound effect on adoptively transferred GD2.CAR-T cells, which have accomplished remission in patients with neuroblastoma . IntratuOne of the strongest barriers to successful T-cell therapy for solid tumors is the expression of inhibitory immune checkpoint ligands expressed on tumor cells  Figure . These lPDL1) enhances the antitumor effect of HER2.CAR-T cells, which were recently reported to be safe in patients with sarcoma  and screened them for their ability to enhance HER2.CAR-T mediated killing of head-and-neck squamous cell carcinoma (HNSCC) targets. We found that only IL-12p70 mediated tumor regression in conjunction with PD-L1 blocking antibody in both HPV positive and negative HNSCC xenograft models. We then generated a single vector expressing IL-12p70 with the PD-L1 blocking antibody which was co-injected with an oncolytic adenoviral vector (CAdVECIL12_PDL1). This treatment combined with HER2.CAR-T cells was able to control both primary and metastasized tumors in an orthotopic model of HNSCC causing lymph node metastasis similar to those seen in HNSCC patients. This superior anti-tumor effect leads to 100% survival of animals treated with the combination of CAR-T cells and CAdVECIL12_PDL1 for more than 120 days without xenogenic GVHD after single treatment  will be needed to understand how host immune responses  contribute to this combinatorial therapy.Armed OVs can be rationally designed to provide T-cells with optimal synergistic molecules for specific tumor targets and therefore represent an ideal platform for targeted cancer therapies. Based on clinical trial data with CAR-T cells for solid tumors, we may be able to identify appropriate molecule(s) expressed by OVs for each CAR construct and for each target tumor/tissue to maximize the anti-tumor effect of CAR-T cells. Since safety of both agents as monotherapy have been demonstrated in numerous clinical trials, and our data indicate that we can obtain durable responses with 1-2 log lower dosages of each agent used as monotherapy in preclinical models , 36, comAll authors listed have made a substantial, direct and intellectual contribution to the work, and approved it for publication.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Recent studies in cancer diagnostics have identified microRNAs (miRNAs) as promising cancer biomarkers. Single nucleotide polymorphisms (SNPs) in miRNA binding sites, seed regions, and coding sequences can help predict breast cancer risk, aggressiveness, response to stimuli, and prognosis. This review also documents significant known miR-SNPs in miRNA biogenesis genes and their effects on gene regulation in breast cancer, taking into account the genetic background and ethnicity of the sampled populations. When applicable, miR-SNPs are evaluated in the context of other patient factors, including mutations, hormonal status, and demographics. Given the power of miR-SNPs to predict patient cancer risk, prognosis, and outcomes, further study of miR-SNPs is warranted to improve efforts towards personalized medicine. Breast cancer is the second most common form of cancer diagnosed in American women, after skin cancer. It is among the most frequently diagnosed cancers worldwide, with 2.08 million new cases in 2018  . These mThe first documented involvement of microRNAs (miRNAs) in human cancers was when miR-15 and miR-16-1 were found to be downregulated in chronic lymphocytic leukemia (CLL) , revealimiRNA expression and targeting can be significantly altered by the presence of single nucleotide polymorphisms (SNPs). SNPs are the most frequently encountered point mutations in the genome, with the first genomic SNP being identified as early as 1976 . Since tmiR-SNPs located in regulatory regions of genes, at miRNA binding sites, or at quantitative trait loci play a major role in gene function and phenotype . Such miSNPs affecting proteins responsible for post-transcriptional processing of miRNAs can significantly affect miRNA function. Briefly, miRNAs go through two rounds of enzymatic processing after initial transcription-processes, which implicate miRNA-specific exonucleases, transport proteins, and signaling cascades . The resDROSHA: The nuclear RNase III, Drosha, is the core nuclease that executes the initiation step of miRNA processing, cleaving primary miRNAs to release hairpin-shaped pre-miRNAs. A case-control study of the SNPs rs644236 (C > T) and rs7737174 (A > G) within the DROSHA gene demonstrated increased risk of developing breast cancer  in Korean post-menopausal women . No. Nop = 0KRAS-variant-associated breast cancer risk appears to be impacted by estrogen withdrawal. McVeigh and colleagues observed that withdrawal from hormone replacement therapy (HRT) was linked to a higher rate of TNBC in KRAS-variant patients (p < 0.001). Furthermore, patients who previously used HRT and had the KRAS-variant tended to be diagnosed with higher grade tumors compared to non-variant patients who had previously been on HRT , indicating that overall the KRAS-variant is predictive of an aggressive tumor biology in patients with low estrogen or experiencing abrupt estrogen withdrawal  + and HR-cases, 3 times as many variant patients had HER2 + (42.9%) versus HER2-(13.3%) tumors, and KRAS-variant tumors had significantly higher histopathological grades . In. InmiR-4Use of miRNA-binding site SNPs as predictive biomarkers holds immense potential as it allows risk stratification for cancer development and cancer biology. This opens up additional avenues of designing efficient prevention strategies, and ensuring cost-effective applications of advanced diagnostic tests . Work re


High low-density lipoprotein cholesterol (LDL-C) levels have been established as a risk factor for fatal cardiovascular (CV) events, including acute coronary syndrome (ACS), whose principal pathogenesis is the disruption of culprit plaques composed of a large lipid core under a thin fibrous cap. This structure, termed thin-cap fibroatheroma (TCFA), is considered vulnerable. Statins or 3-hydroxy-methylglutaryl coenzyme A reductase inhibitors can stabilize vulnerable coronary plaques by lowering LDL-C levels, and this stabilization prevents the occurrence of ACS. Clinical trials involving more than 170,000 participants conducted over two decades have demonstrated the beneficial effects of statins in reducing the risk of CV events by 25%; however, 75% of coronary events manifest despite treatment, possibly because of underlying residual risk . Bec. Bec3]. ApoC-III is present on the surface of TGRLs together with ApoC-II. ApoC-II is an obligate activator of LPL . ConversFibrates, which activate peroxisome proliferator-activated receptor alpha (PPARα), suppress the mRNA expression of ApoC-III and reduce serum ApoC-III levels as well as serum TG and ApoB-48 levels in the fasting and postprandial states. Fibrates are the most effective agents for lowering elevated TGRL levels. Prespecified and post-hoc subgroup analyses in a clinical trial using fenofibrate revealed a significant reduction in CV events in patients with high TG levels and low high-density lipoprotein cholesterol levels. Contrarily, the use of fibrates is frequently associated with adverse effects (AEs), such as an increase in liver enzyme and serum creatinine levels. Recently, pemafibrate, a novel selective PPARα modulator, has been developed. It induces AEs less frequently than fenofibrates . The ongElevated serum ApoC-III levels might impair the postprandial metabolism of TGRLs and reduce the stability of coronary plaques. To prevent the incidence of ACS, reducing serum ApoC-III and TGRL levels in the fasting and postprandial states is an important and effective therapeutic strategy.


Recent evidence has suggested that extracellular microRNAs have crucial roles in intercellular communications and are promising as minimally invasive biomarkers for various diseases including cancers. Oxidative stress also plays an essential role in homeostasis and disease development. This systematic review aims to clarify the current evidence on the interaction between oxidative stress and extracellular microRNAs. We identified 32 studies that provided information regarding the association between oxidative stress and extracellular microRNAs: 9 focused on the central nervous system, 11 focused on cardiovascular diseases, and 4 focused on liver injury. Endothelial cell-specific miR-126-3p was the most studied extracellular miRNA associated with oxidative stress. In addition, we highlight some reports that describe the mechanisms of how oxidative stress affects extracellular microRNA profiles in liver injury. In liver injury, the levels of miR-122-5p, miR-192-5p, miR-223-3p, and miR-1224-5p were reported to be elevated in the sera. The release of miR-122-5p, miR-192-5p, and miR-1224-5p from hepatocytes may be attributed to oxidative stress. miR-223-3p could be released from neutrophils and suppress oxidative stress in the liver. Elucidation of the mechanisms of the interaction between extracellular microRNAs and oxidative stress would improve our pathophysiological understanding as well as future medical practice. They modulate target gene expression at the post-translational level by guiding the RNA-induced silencing complex to miRNA target sites in the 3' untranslated region of mRNAs, leading to mRNA degradation or the inhibition of translation. Currently, 2,588 mature human miRNAs are listed in the miRNA registry (miRBase release 21). Among them, 300–500 miRNAs can be detected as stable extracellular miRNAs in circulation. In addition to those encapsulated within EVs, some extracellular miRNAs are bound to either RNA-binding proteins or high-density lipoproteins. As changes in extracellular miRNA profiles are associated with various disease conditions, they are attractive candidates for minimally invasive biomarkers. Furthermore, studies on extracellular miRNA have revealed functional cell-to-cell miRNA transfer.,12In multicellular organisms, cells can exchange information using single molecules such as hormones and cytokines. In addition, recent evidence has revealed that extracellular vesicles  can transport various molecules, such as nucleic acids, proteins, and lipids, between cells and play pathophysiological roles.Oxidative stress resulting from the increased production or inadequate removal of reactive oxygen species (ROS) plays a key role in the pathogenesis of aging, atherosclerosis, Alzheimer’s disease, cancer, etc. As both extracellular miRNAs and oxidative stress are essential in homeostasis and disease development, knowledge about the interaction between them could provide new physiological insights. Here, we systematically reviewed the literature to clarify the current progress in the field and highlight some notable studies on digestive diseases, which can enhance our understanding of the field and point out future directions for investigation.We searched MEDLINE for studies published until 21 December 2017 using a search strategy (("oxidative stress"[MeSH Terms] OR ("oxidative"[All Fields] AND "stress"[All Fields]) OR "oxidative stress"[All Fields]) AND ("micrornas"[MeSH Terms] OR "micrornas"[All Fields] OR "microrna"[All Fields])) AND (circulating[All Fields] OR ("serum"[MeSH Terms] OR "serum"[All Fields]) OR ("plasma"[MeSH Terms] OR "plasma"[All Fields])) OR ("exosomes"[MeSH Terms] OR "exosomes"[All Fields] OR "exosome"[All Fields]) OR extracellular[All Fields] OR salivary[All Fields] OR ("urinary tract"[MeSH Terms] OR ("urinary"[All Fields] AND "tract"[All Fields]) OR "urinary tract"[All Fields] OR "urinary"[All Fields]) OR juice[All Fields] OR fluid[All Fields] OR ("ascites"[MeSH Terms] OR "ascites"[All Fields]) OR effusion[All Fields] OR ) NOT ("review"[Publication Type] OR "review literature as topic"[MeSH Terms] OR "review"[All Fields]).in silico prediction, and papers that did not focus on human health or diseases.We included English-written original articles that described the association between circulating miRNAs and oxidative stress in their assessment. The exclusion criteria employed are as follows: papers that were not written in English, review articles, papers that only concerned intracellular miRNAs, papers that mentioned the association between circulating miRNAs and oxidative stress only based on –44 These were published after 2013, and the number of studies increased every year. More than half of the identified studies were focused on cardiovascular diseases and/or the central nervous system.We obtained 150 published studies from the initial search, 32 of which were identified as focusing on the association between extracellular miRNAs and oxidative stress   was found to prevent the maximal exercise-induced increase of circulating miR-126-3p in patients with intermittent claudication. Wang et al. reported that miR-126-3p contained in endothelial progenitor cell-derived microvesicles (EPC-MVs) can suppress oxidative stress and promote angiogenesis of endothelial cells via the PI3K/eNOS/NO pathway. They also showed that miR-126-3p was increased in MVs released from EPCs cultured in a serum deprived medium (starvation stress), whereas it was decreased in MVs released from EPCs cultured in a serum-deprived medium containing tumor necrosis factor-α (TNFα) (apoptotic stress). This example clearly showed that oxidative stress can be a regulator of extracellular miRNAs, and can be regulated by extracellular miRNAs.Among these 32 studies, the most frequently documented miRNA was miR-126-3p. As miR-126-3p is located within intron 7 of the EGF-like domain 7 (Egfl7), which is an endothelial-cell-derived secreted peptide, miR-126-3p is specifically expressed in endothelial cells.Regarding the digestive system, 4 studies were focused on liver injury as indicated in Table  Mice lacking the gene encoding miR-122-5p are viable but develop temporally controlled steatohepatitis, fibrosis, and hepatocellular carcinoma. In healthy individuals, the number of exosomes containing miRNA-122-5p significantly increases in the serum after alcohol binge drinking. Exosomes derived from ethanol-treated human hepatoma cells (Huh7.5 cells) are taken up by monocytes/macrophages and Kupffer cells and horizontally transfer miR-122-5p. In monocytes, exosome-transferred miR-122-5p inhibits the HO-1 pathway and increases the levels of pro-inflammatory cytokines such as IL-1β and TNFα. Mosedale et al. reported that early increases in exosomal miR-122-5p tend to be associated with mitochondrial-induced apoptosis and oxidative stress during idiosyncratic drug-induced liver injury. Taken together, alcohol- or drug-induced cellular stress would promote the release of miR-122-5p-containing exosomes in hepatocytes in the absence of overt necrosis. Circulating miR-122-5p could be an early biomarker for damaged hepatocytes.miR-122-5p is one of the most investigated miRNAs; it is abundantly expressed in hepatocytes but is absent or expressed at very low levels in other cell types.et al. discovered that it is also up-regulated in the serum of patients with acute liver failure. Intracellular miR-1224-5p was found to be up-regulated in hepatocytes following in vivo and in vitro ischemia-reperfusion or H2O2 stimulation. They also demonstrated that miR-1224-5p could suppress the anti-apoptotic gene Nfib, leading to impaired proliferation and elevated apoptosis. More importantly, increased serum levels of miR-1224-5p were found to be associated with survival in acute liver failure .Although miR-1224-5p is not a liver-specific miRNA, Roy  Supernatant levels of miR-192-5p were also found to be increased in a hypoxia/reoxygenation model of in vitro hepatocyte injury. However, in contrast to the up-regulation of miR-122-5p and miR-1224-5p in hepatocytes, miR-192-5p was reported to be down-regulated in injured livers in vivo and during H2O2 stimulation in vitro. Functional experiments confirmed the protective effect of miR-192-5p down-regulation in hepatocytes through the increase of a target gene (Zeb2), an important suppressor of apoptosis. Based on these results, the authors suggested that the decrease in intracellular miR-192-5p could be caused by the release of miR-192-5p from hepatocytes during acute liver injury. A limited number of reports also show reciprocal changes in intracellular and extracellular miRNAs, suggesting that some miRNAs might be actively and selectively released from cells in specific conditions.,52The same researchers also reported that serum miR-192-5p levels are selectively elevated in patients with liver injury and closely correlated with serum miR-122-5p levels.et al. found that the serum miR-223-3p levels of alcoholics were elevated compared with those of healthy controls by miRNA microarray analysis, and miR-223-3p could also be a possible biomarker for alcoholic liver injury. However, miR-223-3p was not released from hepatocytes but present at high levels in neutrophils. In mice, the levels of miR-223-3p were found to be increased in both the serum and neutrophils upon ethanol intake. They also showed that miR-223-3p could directly inhibit IL-6 expression and subsequently inhibit p47phox expression in neutrophils. In miR-223-3p-deleted mice, the expression of IL-6 and the phagocytic oxidase p47phox was enhanced in the liver, leading to ROS generation, neutrophil infiltration, and hepatic injury upon ethanol administration. ROS production by neutrophils and ethanol-induced liver injury were suppressed by p47phox deletion. In summary, miR-223-3p in neutrophils could be an important regulator for blocking neutrophil infiltration in alcoholic liver disease.Li In this systematic review, we identified 23 studies indicating that oxidative stress could affect extracellular miRNA profiles and that some transported miRNAs could play cytotoxic or cytoprotective roles in recipient cells. Although a number of studies addressed the use of extracellular miRNAs as biomarkers for various diseases, the regulatory mechanisms of extracellular miRNAs remain unclear. Further studies on oxidative stress should be conducted to shed light on this issue.In the case of digestive diseases, all studies on the association between oxidative stress and extracellular miRNAs were focused on liver injury. In acute liver injury and hepatitis, circulating miRNAs regulated by intrahepatic oxidative stress seem to be powerful assessment tools for determining the extent of liver damage. The most important requirement for the use of a biomarker of acute diseases, such as acute liver injury, acute pancreatitis, and acute myocardial infarction, is to facilitate rapid measurement. Since it takes 1 to 3 days to obtain results by conventional quantitative RT-PCR or microarray, novel methods are required to use circulating miRNAs for the evaluation of acute diseases. In addition, hundreds of previous reports have shown that circulating miRNA signatures are dramatically altered through the carcinogenic process.,54 Nevertheless, none of the studies demonstrate the importance of the association between oxidative stress and extracellular miRNAs in cancer development or progression. As the aberrant miRNA profiles in cancer tissues and their precursor lesions such as Barrett’s esophagus, gastric intestinal metaplasia, or the inflamed colonic mucosa in ulcerative colitis are well-known,–57 extracellular miRNAs released from malignant and pre-malignant lesions should be studied in detail with a focus on oxidative stress to promote cancer prevention and early detection.In digestive carcinogenesis, exposure to oxidative stress plays crucial roles.In conclusion, although there are some well-conducted studies, knowledge of the association between oxidative stress and extracellular miRNAs is rapidly increasing but still limited. Further studies in this area would uncover unique cell-cell interactions and lead to changes in future clinical practice.


Synthesis of the longest heparin-related oligosaccharide backbones is enabled by efficient iterative [4]n-mer homologations via solution-phase synthesis. Pure-Shift HSQC NMR provides a dramatic improvement in anomeric signal resolution. n-mer heparin-backbone oligosaccharides ranging from 16-mer through to the 40-mer in length is described. This demonstrates for the first time the viability of generating long sequence heparanoids by chemical synthesis, via practical solution-phase synthesis. Pure-Shift HSQC NMR provides a dramatic improvement in anomeric signal resolution, allowing full resolution of all 12 anomeric protons and extrapolation to support anomeric integrity of the longer species. A chemically pure 6-O-desfulfated GlcNS-IdoAS icosasaccharide (20-mer) represents the longest pure synthetic heparin-like oligosaccharide.The chemical synthesis of long oligosaccharides remains a major challenge. In particular, the synthesis of glycosaminoglycan (GAG) oligosaccharides belonging to the heparin and heparan sulfate (H/HS) family has been a high profile target, particularly with respect to the longer heparanome. Herein we describe a synthesis of the longest heparin-related oligosaccharide to date and concurrently provide an entry to the longest synthetic oligosaccharides of any type yet reported. Specifically, the iterative construction of a series of [4] The synthesis of long sequence oligosaccharides (≥10 monosaccharide units) is an area of enduring challenge for synthetic carbohydrate chemistry across a diversity of oligosaccharide types. Despite advances in synthetic methods and recent progress in supported and automated chemistry,d-glucosamine and uronic acid units, the latter constituting either d-glucuronic or l-iduronic acid. H/HS typically occurs in oligomers up to 40–50 units in length and though native oligomers range to >150 in length, most biological recognition relates to shorter oligomers, spanning the 6–40-mer range. Additionally, the backbone is modified by a variety of different sulfation patterns, mostly involving the 2-amino and O6 of d-GlcN and the O2 of the uronic acid, with rarer d-GlcN O3 sulfation, and are organized into low- and high-sulfation domains with heparin typified by higher iduronate content and sulfation than heparan sulfate. These oligosaccharides are known to bind to over one hundred proteins and play many diverse regulatory roles in biology, including angiogenesis, inflammation, stem-cell differentiation, pathogen infectivity and are also essentially involved in chemokine biology, regulating oligomerization dynamics and cell-migration properties.Heparin and heparan sulfate (H/HS) are highly polydisperse examples of glycosaminoglycan (GAG) saccharides, structurally characterized by alternating 1 → 4-linked in vitro and in vivo biological consequences of changes in sulfation locations.13The diverse structural microheterogeneity of the natural oligosaccharides and these pervasive regulatory roles ensure that defining the effects of specific structural differences on binding and biological effects of H/HS sequences remains at the forefront of carbohydrate chemical biology, whilst also amongst the most structurally-diverse and functionally-complex of oligosaccharide targets. There is considerable evidence that native H/HS oligosaccharides interact with various heparin-binding proteins across a wide range of oligomer lengths. Recent structural work has shown that even H/HS disaccharides can provide binding selectivity for the important growth factors FGF1 vide infra) several other syntheses of H/HS-related 10-mer and 12-mer oligosaccharides have been described,Synthetic access to chemically pure H/HS species , is believed to involve binding to HS sequence(s) of >20 units.Synthetic access to the longer heparanome backbones has been elusive. A review has reported a 16-merRecent advance in the area of less functionalized GAGs is highlighted by a report on the synthesis of hyaluronic acid, a simpler unsulfated GAG constituting a repeating GlcA-GlcNAc unit, up to a 15-mer.1O-desulfated 12-mer on gram scale,O-sulfated LMWH-like 12-mer.n iterations are likely to become limited in practicability for access to substantively longer sequences due to pathway length, separation convergence and possible cumulative isomerism. We also therefore recently reported the synthesis of a related heparin-like dodecasaccharide by replacing disaccharide iteration with a GlcN-IdoA-GlcN-IdoA tetrasaccharide block approach, exploiting the utility of iduronate thioglycoside donor capability at the tetrasaccharide, rather than disaccharide, level,n iterations, coupled with our multi-gram scale access to tetrasaccharide modules, suggested a +[4]n block iteration strategy might provide a viable homologation to substantially longer GAG sequences, which have not been addressable hitherto. Block synthesis has proven effective in a number of elegant branched oligosaccharide syntheses,n heparin-like [GlcN-IdoA]n backbone oligosaccharides in the 20 to 40-mer range, which had previously eluded synthetic access. This more than doubles the length of heparin-like oligosaccharides which have previously been amenable to chemical synthesis, and also provides the longest synthetic oligosaccharides  of any type yet reported by solution or by solid-supported methods, exceeding the recent advance reported for small scale provision of a uniform non-GAG 30-mer oligomannoside,We recently described a scalable iduronate synthesis facilitating multi-gram access to GlcN-IdoA disaccharides suitable for oligosaccharide homologation,2 tetrasaccharide donor 1 We sought to evaluate the use of a [GlcN-IdoA]It was also important to demonstrate that this route to extended oligosaccharides could deliver usable quantities of materials with reproducible homologation efficiencies and anomeric integrity throughout the sequence. For all oligosaccharide synthetic routes the reproducibility of coupling efficiency (and anomeric integrity) for longer chains is a recurrent challenge. Similarly, convergence of separability of product from incompletely glycosylated long acceptors presents difficulties as the sequences become longer. We reasoned that application of our block strategy could assist both aspects, by significantly reducing the number of sequential steps (fewer glycosylations and decreasing cycles for any accumulating loss of anomeric coupling integrity) and providing a more significant structural change upon each homologation step (thus aiding separation and purification).1 2 3 in very high yield  and with high anomeric selectivity  oligomannoside.This establishes a chemical synthesis of the longest synthetic oligosaccharides of any structural type yet reported and also significantly increases the length of synthetic heparanoid oligosaccharides now accessible by chemical synthesis. This provides a paradigm-shift in planning access to the longer heparanome by demonstrating an effective solution-phase strategy that can both span the entire target length range of likely biological relevance and do so on viable scales.+ as the base peak for all species, illustrated in Mass spectral data for the series of oligosaccharides provided convincing supporting data for structure, with molecular ions present for [M + Na]3) resulting in faster correlation times and therefore sharper linewidths.§The poorer linewidths and dispersion in a deprotected, aqueous sample of the 12-mer precluded resolution of all the anomeric signals, even using NUS-HSQC. However, to record 2D spectra with sufficient acquisition time in the indirect dimension to resolve all the peaks would have taken a prohibitively long time, so non-uniform sampling (NUS)1H13C-HSQC spectra was employed. Resolution was improved further by homonuclear decoupling during acquisition using a BIRD pulse trainWe wished to use NMR to confirm the anomeric integrity across this series. Significant resolution enhancement of the twelve anomeric resonances for the 12-mer we reasoned may then facilitate extrapolation to provide sufficient resolution improvement for the longer series, important for defining anomeric integrity in the longer sequence syntheses. NMR confirmation of the purity of the final products was attempted prior to deprotection of the sugar hydroxyls, the protecting aromatic groups providing extra dispersion through ring current shifts, and also low viscosity organic solvent  was saponified, O-sulfated and hydrogenatedO-sulfated, N- and 6-O-desulfated icosasaccharide amine (A4I2-[A0I2]9)-OMe5018 -OMe 18 was then N-sulfated to provide the IdoA 2-O- and glucosamine N-sulfated (S4I2-[S0I2]9)-OMe 20-mer 19 in 93% yield. Shift of H2 signals confirm no remaining NH2 sites  which has significance for the current importance of developing pure synthetic heparin-type replacements. Additionally, this report describes an important advance in NMR structural assignment at 12-mer level and quantification of anomeric integrity in long oligosaccharides. NUS Pure-Shift HSQC is shown to provide very dramatic resolution enhancement and was applied to (i) provide the first clear resolution of the 12 anomeric signals for a synthetic heparin-related dodecasaccharide and (ii) thereby allowed comparative data up to a synthetic 36-mer providing direct, robust NMR support regarding the anomeric integrity in these very long synthetic oligosaccharides. This will have value for the analysis of other long synthetic oligosaccharide systems.The first purely synthetic chemical approach to provide native-backbone heparin-like oligosaccharides in the longer heparanome (>16-mer) is reported. This demonstrates that a tetrasaccharide block-wise iterative approach enables syntheses up to and including the longest synthetic oligosaccharide (of any type) yet described (40-mer). This now facilitates chemical synthesis to this previously inaccessible area of glycospace with sequence homogeneity. This work shows that adapting solution phase synthesis using block-synthetic modules can provide a practicable route to long saccharides of high purity and stereochemical integrity. This report also describes the chemical synthesis of a pure heparin-like Supplementary informationClick here for additional data file.