Mass application of SARS-CoV-2 vaccines is a major approach to reduce SARS-CoV-2 transmission and COVID-19 symptoms worldwide. The inactivated SARS-CoV-2 vaccine is one of the most widely administered vaccine types in many countries, with over 2 billion doses administered as of 31 August 2021. However, clinical trials estimate 65%–85% protection from detectable symptoms for inactivated SARS-CoV-2 vaccines , with 15et al. analyzed in vitro neutralizing antibodies with clinical protection data in seven vaccine clinical trials and estimated 50% protection against detectable infection to be 20.2% of the mean convalescent neutralizing antibody level  and interleukin-2 [IL-2] secreting T cells upon SARS-CoV-2 peptide pool stimulation) before receiving the third dose despite insufficient antibody responses among these people Fig. F. It hasTo further dissect the T cell responses in CD4+ and CD8+ cells, we performed activation-induced marker (AIM) assays [In the safety assessment, we found that systemic muscle pain and headaches tended to be less common after the third dose compared to the first two doses . Conversely, local (injection-site) adverse reactions were generally more common after the third dose than the first two doses . No grade three or four events were reported in our third-dose cohort.Many countries are launching mass application of the third dose as a ‘booster’ dose, but whether a third dose elicits an adequate antibody response among non-responders is unknown. Our results confirmed the value of a third dose among non-responders, as most non-responders gained neutralizing antibodies, reaching seropositive status. However, the antibody level elicited by a third dose in this special population is not adequate for neutralizing the Delta variant. The weakened protection against this variant of concern in non-responders is likely due to their defective antibody response compared to good-responders . Thus, uWe observe that neutralization against the Delta variant is linearly correlated with the neutralizing antibody titer of the wild-type virus . Thus, we anticipate that a third dose immunization with stronger immunogenicity  will likely provide adequate protection against the variant virus in non-responders. In sum, we conclude that a third dose of inactivated vaccine should be considered to ensure that most of the vaccinated population acquire sufficient humoral immunity, and trials with sequential vaccination using different types of vaccine, particularly vaccines targeting variants of concern, are encouraged in order to achieve a stronger antibody response for protection against SARS-CoV-2 variants. This study is limited by its relatively small sample size and therefore cannot completely explore the mechanism behind the inter-individual variation of immune response to a third dose among non-responders. Future studies with a larger cohort will likely provide deeper mechanistic insights.nwac066_Supplemental_FilesClick here for additional data file.


The most severe clinical manifestations of the horrifying COVID-19 disease, that claimed millions of lives during the pandemic time, were Acute respiratory distress syndrome (ARDS), Coagulopathies, septic shock leading eventually to death. ARDS was a consequence of Cytokine storm. The viral SARS-COV2infection lead to avalanche of cytokines and eicosanoids causing “cytokine storm” and “eicosanoid storm.” Cytokine storm is one of the macrophage-derived inflammatory responses triggered by binding of virus particles to ACE2 receptors of alveolar macrophages, arise mainly due to over production of various pro-inflammatory mediators like cytokines, e.g., interleukin (IL)-1, IL-2, and tumor necrosis factor (TNF)- α, causing pulmonary edema, acute respiratory distress, and multi-organ failure. Cytokine storm was regarded as the predictor of severity of the disease and was deemed one of the causes of the high mortality rates due to the COVID-19. The basis of cytokine storm is imbalanced switching between an inflammation increasing - pro-inflammatory (M1) and an inflammation regulating-anti-inflammatory (M2) forms of alveolar macrophages which further deteriorates if opportunistic secondary bacterial infections prevail in the lungs. Lack of sufficient knowledge regarding the virus and its influence on co-morbidities, clinical treatment of the diseases included exorbitant use of antibiotics to mitigate secondary bacterial infections, which led to the unwarranted development of multidrug resistance (MDR) among the population across the globe. Antimicrobial resistance (AMR) needs to be addressed from various perspectives as it may deprive future generations of the basic health immunity. Specialized pro-resolving mediators (SPMs) are generated from the stereoselective enzymatic conversions of essential fatty acids that serve as immune resolvents in controlling acute inflammatory responses. SPMs facilitate the clearance of injured tissue and cell debris, the removal of pathogens, and augment the concentration of anti-inflammatory lipid mediators. The SPMs, e.g., lipoxins, protectins, and resolvins have been implicated in exerting inhibitory influence on with cytokine storm. Experimental evidence suggests that SPMS lower antibiotic requirement. Therefore, in this review potential roles of SPMs in enhancing macrophage polarization, triggering immunological functions, hastening inflammation resolution, subsiding cytokine storm and decreasing antibiotic requirement that can reduce AMR load are discussed.  At the time of publication of this article a total of 113 million individuals have been clinically affected by the current pandemic. Of these, approximately 89 million have recovered, 21 million still have not and more than 2 million lives have been lost to the disease. The causative organism for the coronavirus disease 2019 (COVID-19) is the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2), which is an enveloped, single-stranded, positive-sense RNA virus, that belongs to the group of Beta coronaviruses 2b lineage of the family coronaviridae, and the order Nidovirales . It closStreptococcus pyogenes, Neisseria meningitidis, Moraxella catarrhalis, Streptococcus pneumoniae, Haemophilus influenzae, and Staphylococcus aureus are all capable of causing influenza-related infections , as part of “cytokine storms” . Cell deAlveolar macrophages may be integral to COVID-19 disease progression and disease-induced deaths. Infection of lungs by SARS-CoV-2 virus followed by failure to rapidly remove the virus leads to severe inflammatory response, tissue damage and fibrosis. Prolonged overactivation of macrophages and increased release of pro-inflammatory cytokines result in clinical manifestations that are markedly identical to those of macrophage activation syndrome (MAS). This being the case, identifying methods to regulate macrophage response in cases of severe COVID-19 is key to recovery . In thisvia diagnostic testing. Another reason is secondary infections associated with COVID-19, where antimicrobial use is required to kill invading bacteria and fungi  emerged as a notable issue quite a while before the start of the COVID-19 pandemic. With the influx of novel antibiotics at a decline since the 1980s, instances of reported cases of infections by drug-resistant organisms have soared . COVID-1nd fungi . Besidesnd fungi . These snd fungi . OverallStaphylococcus aureus, Streptococcus pneumoniae, Neisseria meningitides, Haemophilus influenzae, Klebsiella pneumoniae, and members of the genus Proteus, Enterobacter, and Citrobacter species. These are present in the hospital setting, and their spread is nosocomial , n-3 docosapentanoic acid and docosahexaenoic acid   are endogenous lipid mediators, produced from innate immune cells, formed C22:6–3) . The difC22:6–3) . FurtherC22:6–3) . FinallyC22:6–3) . SPMs seC22:6–3) .Pseudomonas aeruginosa induced pneumonia  and production of IL-12 takes place during infection by Toxoplasma gondii . Lipoxintenuated  as depictenuated . Administenuated , Plasmodtenuated  and Tryptenuated .Macrophages (MΦ) are heterogenous family of innate immune cells . MΦs areSARS-COV2 caused disastrous COVID-19 pandemic that claimed several millions of lives. The overwhelmed death rate associated with COVID-19 was mainly attributed to acute respiratory distress syndrome (ARDS), cytokine storm and fatal multi-organ failure. Other pathological events observed in COVID-19 cases are disseminated intravascular coagulation (DIC) syndrome, edema, and pneumonia, which are prime consequences of macrophage activation syndrome (MAS) . Angiotevia clearing pulmonary surfactant and cellular debris, essential for tissue repair and resolution of inflammation  specifically TLR7/8 is an invader sensing receptor able to recognize ssRNA sequences of SARS –COV2, this recognition could stimulate M1 pro-inflammatory macrophages which may trigger a rapid type I IFNs(interferon) response, consequently causing innate immune hyperactivation, aberrant cytokine production and ultimately lead to acute lung injury (ALI), ARDS, multi-organ fibrosis and dysfunction . Elevatevia the IL-23/IL-17A pathway (Lipoxins (LXs), can inhibition of viral entry and its replication, downregulate ACE2 receptors, inhibit release of the pro-inflammatory cytokines. Moreover, LXs can augment the polarization of macrophages from (M1) to (M2) phenotype, causing a shift from pro-inflammatory to anti-inflammatory environment, thereby enhancing the inflammation resolving actions of macrophages in SARS-CoV-2 infection . RvE1and pathway .via potentiating MΦ phagocytosis, decreasing release of pro-inflammatory cytokines and chemokines from MΦ, in response to the viral spike protein  was enhanced due to overexpression of miR-466l in macrophages, which ultimately enhanced resolution of inflammation, efficiently demonstrated in mice models by Li et al. . RvD1 an protein . Maresin protein . Maresin protein . RvD6 is protein . The pro protein .SARS-CoV-2 triggers cytokine production in overabundance, resulting in exaggerated inflammatory response and an avalanche of cytokines, dubbed “cytokine storm” (CS) or “quasi cytokine storm” in patients with COVID-19 . CS can Pro-resolving mediators such as SPMs can be used to counter CS in cases of severe COVID-19 . ResolviThe major reason for severity of COVID-19 infection is attributed to the acute uncontrolled inflammatory response in the lungs known as cytokine storm. The macrophages play important role in regulating COVID-19 inflammation by switching between two forms: pro-inflammatory macrophages (M1) and anti-inflammatory macrophages (M2) forms. Since the lungs are preferred target of most respiratory viruses including COVID-19 so the involvement of lung alveolar macrophages in progression of severe SARS-COV 2 is eminent. ACE2 receptors are essential for entry of SARS-COV2, and are found to be expressed on alveolar macrophages (AMs). The shifting of alveolar macrophages from M2 to M1 form triggers the SARS-COV 2 virus to smoothly infect the lungs, replicate and consequently induce cytokine storm. This causes build-up of oedema i.e. accumulation of inflammatory fluids in lungs that averts oxygen transfer from lungs towards blood capillaries inducing hypoxia and if untreated then associated multiple organ failure. The specialized pro-resolving lipid mediators are found to be highly effective in inhibiting inflammatory responses during infection. They are produced by innate immune cells from essential long chain fatty acids and are found to improve the concentration of anti-inflammatory lipid mediators. The specialized pro resolving lipid mediators such as protectins, lipoxins and resolvins have been strongly associated with inhibition of cytokine avalanche or cytokine storm which is a well-known reason behind COVID-19 deaths. The viral diseases have been always difficult to treat due to numerous factors such as limited anti-viral pharmaceutical agents, frequent re-infections, frequent mutations in RNA viruses and restructuring of genome etc. but SPM’s offers a different approach of treatment wherein we can control severity of infection by controlling inflammation. This is well justified by the fact that most viral infections subside on their own after completion of their cycles in host. Infact the majority of Covid patients developed mild symptoms possibly due to absence of inflammatory responses as cytokine storm. So in near future regulating the respiratory inflammation shall be the goal in order to avoid progression to severe respiratory distress. Thus there is great scope of using SPM’s against COVID-19 variants and also other viral infections in near future particularly RNA viruses which evades novel pharmaceutical agents due to their mutagenic capabilities.


Public health authorities around the world have been disseminating messages to support mental health and psychosocial well-being during the COVID-19 pandemic. Based on the Portuguese guidelines, we have developed the Adherence Scale to the Recommendations for Mental Health during the COVID-19 Pandemic (ASR-MH-COVID19) to better understand this health behaviour.To analyse the relationship between sociodemographics, personality traits, Adherence  and psychological distress.413 individuals  completed an on-line survey, in September-December 2020, including sociodemographic questions, ASR-MH-COVID19, NEO-FFI-20 and Depression Anxiety Stress Scale (DASS-21) and Health Perception Scale.Adherence scores did not significantly differ by gender, age and years of education. Women presented higher DASS and Neuroticism scores (p<.01). Adherence were negatively correlated with Neuroticism (r=-.247) and with Depression/Anxiety/Stress , positively with Openness to Experience (r=.174), Conscientiousness (r=.194) and Perceived Health  . Mediation analysis (Hays’ Macro Process - Model 4) revealed that Adherence is a partial mediator between Openness and DASS and Conscientiousness and DASS; when considering Neuroticism, only the direct effect was significant. The effect of Perceived Health  on DASS was also mediated by Adherence.The Health Behaviour Model proposes a pathway linking personality and health that applies to these results about adherence and psychological distress during the COVID-19 pandemic. Personality and perceived health  influence both adherence to mental health behaviours and psychological distress. Understanding personality is vital for health care providers.No significant relationships.


While the fraternity continues to ponder on the mechanisms by which coronavirusdisease (COVID-19) positivity affects the outcome of cardiac surgical subset, weput forth a 3H (Hypoxia-Hemolysis-Hyperinflammation) trilogy aimed atelucidating the liaison between cardiopulmonary bypass  and COVID-19 infection. A sound comprehension of thesame can doubtlessly assist the perioperative team in staging a well-directedpathophysiology-driven management approach. Needless to say, the aforementioned intensifiesthe pre-existing challenges with every likelihood of this trinity to be at the heartof the resultant organ dysfunction and subsequent morbidity and mortality inCOVID-19 positive cardiac surgical cohort.,cardiopulmonary bypass (CPB) associated microcirculatory alterations (inbackground of a non-pulsatile perfusion) are expected to be rather poorlytolerated at the tissue level despite an improved and controlled oxygenationon CPB. Moreover,CPB-related hemodilution makes the matter even worse. Whileprolonged aorticcross-clamp and CPB times translate into ischemic tissue burden, theconcerns about circulatory arrest (mandatory for complex open-heartsurgeries) become manifold[5,.Given a prevailing hypoxemic milieu and an inadequate tissue oxygenationinthe setting of COVID-19-related acute respiratory distress syndrome(ARDS). On the other hand, plausible viral-erythrocyticinteractions are equally conducive to an aggravated red blood corpuscularlysis. Thepresent understanding of these interactions reveals that ACE2, CD26 andCD147 erythrocytic receptors serve as potential attachment sites forSARS-CoV-2 coupling, promoting hemolysis in turn by allowing the virus tomount an attack on the 1-beta haemoglobin chain. At the same time, thehepcidin-mimicking attribute of SARS-CoV-2 augments the level of circulatingtissue ferritin alongside serum iron deficiency and anemia.On one hand, mechanical shear stress and exposure to non-endothelial CPBsurfaces predispose to hemolysis[9,. Focusing on the conduct of CPB in COVID-19 infectedpatients, the inexorable systemic inflammatory response to CPB[10- and the concomitant ischemia-reperfusioninjury can compound the ongoing SARS-CoV-2 cytokine storm with potentiallydetrimental outcomes[13,.Stage III or the hyperinflammatory phase of COVID-19 infection is associatedwith a significant immuno-inflammatory response or cytokine storm with amarked elevation of inflammatory markers such as interleukin-6 (IL-6),C-reactive protein (CRP), lactate dehydrogenase (LDH), tumour necrosisfactor alpha (TNF-α), etc.. In turn,the inflammatory endothelial dysfunction or endothelitis, particularly in thepulmonary microvasculature, accentuates hypoxemia[16,. Despite a widespread endothelitis at the cornerstone ofsystemic thrombotic consequences, description of an overwhelming thrombosis inthe lungs compared to the whole body in COVID-19 patients has paved the way fora neoteric proposition of coining this underlying phenomenon as a ‘pulmonaryintravascular coagulopathy’ in contrast to the usual nomenclature ofdisseminated intravascular coagulation (DIC). In addition to the thrombotic sequelae, theinflammatory contribution to pulmonary arterial hypertension also adds to thehypoxic predilection. Withrespect to the Hemolysis-Hyperinflammation crosstalk, the hyperferritenemia andthe resultant ferroptosis lead to a considerable oxidative stress that canpotentially exasperate the prevailing systemic inflammatory state.While the anemic hypoxia owing to hemolysis understandably perpetuates theongoing hypoxic consequences staging obvious interactions between theHemolysis-Hypoxia facets of the 3H liaison, other intricate molecular mechanismsassociate the Hypoxia-Hyperinflammation facets wherein hypoxia-inducible factor(HIF) is closely linked to the nuclear factor kappa B (NFκB) inflammatorypathway.Moreover, establishment of a surgical airway requires additional precautions and protective measures to avoid aerosolization,such as use of personal protective equipment (PPE) kit, advancement ofendotracheal tube before puncturing the cricothyroid membrane, use of a cuffedtube, holding ventilation (if possible) when the trachea is open and compulsoryuse of a heat and moisture exchange filter[21,.This is aggravated in the cardiac surgical set-up owing to a prolongedpost-operative mechanical ventilation and frequent requirement of suctioning,increasing the risk of disease spread.Health care professionals currently face peculiar challenges in the operatingroom and intensive care unit. Establishment of a definitive airway, suctioning,noninvasive and positive pressure mask ventilation, and supraglottic airwaydevices are considered risk factors for high aerosol production during surgeryand post-operative care. Similarly, an increased systemicinflammatory response, endothelial dysfunction and coagulopathy inexorablyassociated with cardiac surgical patient subset become even more relevant inCOVID-19 patients (pre-existing hyperinflammation and hypercoagulopathy)aggravating the risk of end-organ dysfunction such as stroke and bleedingdiathesis[24,.On the other hand, the application of positive end-expiratory pressure commonlyemployed in the treatment of COVID-19 patients can negatively interact inpatients after heart surgery, impairing right ventricular output andaccentuating left ventricular diastolic dysfunction.Therefore, stratifying patients according to disease acuity, time permitted forpreoperative testing for COVID-19, a judicial use of personal protectiveequipment kits and precautions to minimizing the exposure risk to the healthcare system has been advised by the American Heart Association and the AmericanCollege of Surgeons amidst this fearsome pandemic era in a leading cardiothoracic and vascularanesthesia journal where they even consider a life-saving modality of extracorporealmembrane oxygenation (ECMO) in COVID-19 as anunhappy marriage of endothelialdysfunction and hemostatic derangements.In the context of the ongoing viral pandemic, it is imperative to reconsider astrengthened and well-aligned perioperative anti-inflammatory armamentarium,rheological preservation on CPB closely backed by sophisticated tissue oxygenationmonitoring, improved organ perfusion and ultrafiltration strategies on CPB, and theincorporation of advancements in CPB such as biocompatible circuits withminiaturized designs, in order to mount a concerted endeavour to combat the unholytrinity of Hypoxia-Hemolysis-Hyperinflammation in our high-risk cardiac surgicalpatients suffering from COVID-19. The importance of the aforementioned discussion isheralded in the title of an Editorial by Seelhammer etal.


With the advent of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas) mediated genome editing, crop improvement has progressed significantly in recent years. In this genome editing tool, CRISPR-associated Cas nucleases are restricted to their target of DNA by their preferred protospacer adjacent motifs (PAMs). A number of CRISPR-Cas variants have been developed e.g. CRISPR-Cas9, -Cas12a and -Cas12b, with different PAM requirements. In this mini-review, we briefly explain the components of the CRISPR-based genome editing tool for crop improvement. Moreover, we intend to highlight the information on the latest development and breakthrough in CRISPR technology, with a focus on a comparison of major variants  to the newly developed CRISPR-SpRY that have nearly PAM-less genome editing ability. Additionally, we briefly explain the application of CRISPR technology in the improvement of cultivated grasses with regard to biotic and abiotic stress tolerance as well as improving the quality and yield. Since the advent of agriculture, plants have been cultivated and utilized as a source of food and energy to feed humans and livestock. The world’s population is expected to increase by 40% in 2050 and the demand for food will be increased by 50% . To meetStreptococcus pyogenes is the most commonly used system and it requires an NGG  PAM sequence for DNA targeting , maize (Zea mays), barley (Hordeum vulgare) and wheat (Triticum aestivum) is explained briefly. Therefore, this review showed the application and usefulness of genome editing as a new breeding technology for crop improvement.This review discusses the components of CRISPR-based genome editing tools in detail and presents a quick comparison of previous main CRISPR variants with the latest one based on PAM sequence requirements. Moreover, an overview of different applications of CRISPR-based genome editing tools is also covered in a schematic diagram and its application for the improvement of agronomic traits as well as biotic and abiotic stresses in cultivated grasses such as rice  lobe and the nuclease (NUC) lobe. The REC lobe has two multi-helix domains, named REC1 and REC2, essential components to bind with the guide RNA and target DNA . REC1 isGuide RNA is composed of two elements, CRISPR RNA (crRNA) and trans-activating CRISPR RNA (tracrRNA). The crRNA is a long sequence of 18–20 base pairs that recognize and specify the target DNA by binding with it . The traBiological systems of CRISPR/Cas are a part of the adaptive immune system of bacteria and archaea, protecting them from nucleic acid invaders such as viruses by cleaving the alien DNA in a sequence-dependent fashion . At the Upon finding the appropriate PAM for the target site, Cas-9 triggers local DNA melting followed by hybridization of RNA with DNA leading to the activation of Cas-9 protein to cleave target DNA . This prAfter DSB, the cellular DNA repair pathways commence. The repair mechanism induces the cell to undertake homology-directed repair (HDR), microhomology-mediated end joining (MMEJ) or non-homologous end joining (NHEJ). HDR occurrence leads towards specific editing by a repair template-specific desired genomic modification . HDR-basStreptococcus pyogenes (SpCas9) needs a GC-enriched site PAM in the form of NGG (N = A/T/G/C) which limits the targeting flexibility. The presence of PAM restrains access to some potential sites due to which many precision edits of targeting sites encoding non-canonical PAM remained inaccessible  and Adenine Base editing (ABE) have been optimized for base editing in rice, wheat, and maize . But, thA vector .These outcomes and unclear shortcomings compel further investigation of structural engineering and application in different systems notably such self-editing was not reported/observed for single base editing in human cells using SpRY-ABEs .Plants are exposed to different environmental stresses such as microbes or climatic changes which are referred to as biotic and abiotic stresses, respectively. Both of these (biotic and abiotic) cause almost 50% yield loss globally . The forOsARM1 and OsNramp5 genes using CRISPR-based gene editing (TaDREB2 (dehydration responsive element binding protein 2) and TaERF3 (ethylene-responsive factor 3) have been successfully targeted in the protoplast using CRISPR-Cas , flooding (hypoxia), salinity, heavy metals, temperature (hot and cold), and their interactions are the major factors hindering agricultural production. Using a variety of breeding approaches, different genes/pathways and regulatory networks involved in stress responses have been determined. To epitome, CRISPR-based genome editing has broadened the target of the biologists to activate or suppress the targeted genes involved in plant abiotic stress resistance . For exa editing . In wheaISPR-Cas . SimilarISPR-Cas . An elevISPR-Cas . Likewis mutants . CRISPR- in rice . Based o in rice , so noveTAMLOA1, TAMLOA2, and TAMLOA3 genes in the wheat genome using the CRISPR-Cas 9 system. In barley plants, the HvMORC1 gene is silenced via CRISPR-Cas 9 which led to an increase in the resistance against Fusarium graminearum  and increased panicle branching (Micro RNA (miRNA) are small RNA molecules with complementary binding sites in target mRNAs, representing a promising avenue to control complex traits since miRNAs can precisely down-regulate any number of co-expressed target transcripts and their respective pathways . Interesranching . Furtherranching . Taken tranching . MoreoveCRISPR-Cas represents the most recent development in genome engineering which has revolutionized crop breeding since 2013. With CRISPR-Cas, genome editing has become a relatively simple, low-cost, and robust process, resulting in huge advances in crop improvement.Producing disease-resistant and environment-adapted crops, as well as improving yields and quality, are the main applications of CRISPR technology in agriculture. One noteworthy example is that preassembled CRISPR-Cas9 ribonucleoproteins were delivered DNA-free into plant protoplasts of rice and wheat . Thus, cin vivo directed evolution efforts to improve other plant characteristics against resistance for high yielding of important agronomic traits to ensure sustainable food security.CRISPR/Cas as a powerful tool led to tremendous advances in crop improvement through precise knockout, knock-in, replacement, point mutations, and fine-tuning of any gene. A potential method to attenuate on-target editing and circumvent the vector self-editing, which gives rise to in activation, off-targeting, should be further explored with a clearer understanding of the characteristics underlying SpRY. Although Cas9 is not capable of PAM-less editing, however, it proved itself better than SpRY for canonical NGG PAM site editing, which reveals the unterminated importance of Cas9 . To date


O-GlcNAc is a pleotropic, enigmatic post-translational modification (PTM). This PTM modifies thousands of proteins differentially across tissue types and regulates diverse cellular signaling processes. O-GlcNAc is implicated in numerous diseases, and the advent of O-GlcNAc perturbation as a novel class of therapeutic underscores the importance of identifying and quantifying the O-GlcNAc modified proteome. Here, we review recent advances in mass spectrometry-based proteomics that will be critical in elucidating the role of this unique glycosylation system in health and disease. Clinical Trial Registration:ClinicalTrials.gov, Identifier NCT05063539Trichoplax adhaerens, the simplest known animal  is a poorly understood post-translational modification (PTM). The modification impacts thousands of distinct sites differentially across cell and tissue types . Canonicn animal . Moreoven animal  The consAlthough identified in the early 1980s, O-GlcNAc remains challenging to study . The regO-GlcNAc is pleiotropic, influencing many biological processes. The efficiency of detained intron splicing inversely correlates with low O-GlcNAc levels while other splicing mechanisms remain unchanged . Pharmacin vivo and patient-derived samples will be particularly important to investigate the role of this system in health and disease. This may become especially apparent with the advent of OGA inhibitors as a novel class of therapeutic .As evident, strategies that perturb OGT and/or OGA are instrumental when probing general phenotypic questions; however, the pleiotropic nature of this system makes mechanistic questions difficult to decipher with genetic knockouts or small molecule inhibitors. As our understanding of this system further develops, methods to map the specific sites of O-GlcNAcylation will be critical to elucidate mechanistic understanding. Techniques that can be applied to Mass spectrometric techniques to map the O-GlcNAc modified proteome and how glycosylation patterns change under different conditions could be powerful platform to assess the impact of O-GlcNAcylation. These strategies provide insight on the specific cellular signaling mechanisms regulated by OGT/OGA. Moreover, mapping specific O-GlcNAc sites provides candidates downstream of OGT/OGA that may be more amenable to perturbation and/or mutation. Typically, mass spectrometry-based proteomics requires high performance liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) to identify and quantify peptides and proteins (reviewed in ). VariouMapping PTMs using DDA methods may require enrichment of the modified peptides or proteins, depending on the stoichiometric abundance of the modification [reviewed in ]. UnmodiLC-MS/MS peptide sequencing techniques fragment precursor ions before the secondary data acquisition event. These product ions and the mass differences between them are used to deduce the peptide identity. For discovery DDA proteomics, the most typical fragmentation techniques employed are collisional-induced dissociation (CID) and higher-energy collisional dissociation (HCD). CID and HCD preferentially break the weakest bonds of the precursor ion. GlcNAc is connected to serine/threonine residues via a labile glycosidic bond . SubsequAs previously noted, mapping the O-GlcNAc modified proteome poses a somewhat unique challenge in traditional bottom-up DDA proteomics, not only due to challenges with site localization when using collisional activation MS2 fragmentation techniques but also to limitations in sample enrichment. O-GlcNAc enrichment has a repertoire of diverse strategies, each with their own advantages and limitations. O-GlcNAc metabolic labeling techniques demonstrate success in numerous iterations, providing a remarkable platform to investigate diverse questions,  but cannThe labile glycosidic bond between a GlcNAc moiety and a serine/threonine residue necessitates innovative mass spectrometry data acquisition techniques. ETD was a key technological advancement for O-GlcNAc site specific mapping . Some frProteomic strategies to elucidate the biological functions of O-GlcNAc signaling will require both accurate site localization and robust quantification. Numerous strategies exist that allow for the quantitative analysis of multiple samples simultaneously, including but not limited to: isobaric mass tags  , SILAC , stable-As mentioned previously, enrichment specificity is important when identifying putative O-GlcNAc sites. N-acetylglucosamine has stereoisomers. N-acetylgalactoseamine is one such stereoisomer that can also modify serine and threonine residues . This makes it challenging to distinguish these entities from their mass addition to fragment peaks alone. Consequently, putative O-GlcNAc sites are reported as HexNAc sites, highlighting stereoisomer ambiguity. O-GalNAc typically forms the basis of branched O-Glycans (reviewed in ), but it. As highlighted by HexNAc modifications have distinctive fragment ions with HCD/CID methods. HexNAc collisional activation produces a characteristic HexNAc oxonium ion peak at 204.086 m/z . In beamThe global profiling of PTMs to investigate their function in health and disease is uniquely suited to mass spectrometry-based proteomics, as neither PTM localization nor stoichiometric abundance is genetically encoded. Bottom-up, DDA proteomic strategies localize and quantify the phosphoproteome  reviewe), the acRelative to other organs, the brain is heavily O-GlcNAcylated . NeuronaClinicalTrials.gov Identifier: NCT05063539). Alectos Therapeutics Inc. in partnership with Merck have completed phase 1 clinical trials for the OGA small molecule inhibitor MK-8719 (Alzheimer’s disease (AD) is the leading cause of dementia. The hallmarks of AD include amyloid plaques and neurofibrillary tau tangles. Tau is a microtubule binding protein that regulates the stability of the neuronal cytoskeleton. In AD, tau is abnormally phosphorylated, and accumulates into toxic neurofibrillary tangles. . Aberran MK-8719 . Global Aberrant O-GlcNAcylation is implicated in numerous cancer types and cancer associated biological processes  reviewe). GlobalBeyond identification and abundance, important questions remain on the functional impact of O-GlcNAcylation on target proteins. One cancer related phenomenon influenced by O-GlcNAcylation is the Warburg effect . The Warin vitro applications.To understand protein-protein interactions mediated by PGK1 T255 O-GlcNAcylation, O-GlcNDAz mediated photo-crosslinking followed by mass spectrometry could identify novel PGK1 mitochondrial interactors. GlcNDAz is an analog of GlcNAc in which the N-acyl position of GlcNAc is replaced with a photo-crosslinker. In cells expressing a mutant form (F383G) of UDP-GlcNAc pyrophosphorylase, a cell-permeable diazirine-modified form of GlcNAc-1-phosphate (GlcNDAz-1-P) is converted to UDP-GlcNDAz. OGT can transfer GlcNDAz from UDP-GlcNDAz to produce O-GlcNDAz modified PGK1, while not dramatically affecting the normal O-GlcNAcylation process. Cells are subsequently irradiated using UV light to activate the diazirine crosslinker, leading to covalent linkage between O-GlcNDAz modified PGK1 and neighboring molecules. The GlcNDAz modified substrate (PGK1) would then be immunoprecipitated, with a subsequent LC-MS/MS follow-up to determine interacting partners . This stThe capability to confidently map the O-GlcNAc modified proteome at the site-specific level has accelerated due to key advancements. O-GlcNAc enrichment strategies are numerous, poising their own advantages and disadvantages. ETD data acquisition strategies are landmark techniques for HexNAc site localization. In conjunction with product-triggered methods, both HCD and ETD fragmentation methods are leveraged to acquire relevant information of putative HexNAc modified precursors. HCD fragmentation may provide a “O-GlcNAc fingerprint” pattern through the dissociation of the HexNAc oxonium ion. This stereoisomer resolution will be critical for investigations that probe O-GlcNAcylation beyond cytoplasmic and nuclear proteins. HCD-pdt-ETD can leverage this information, providing TMT quantitation and HexNAc stereoisomer information in the HCD scan and HexNAc site localization in the ETD scan. Together, advancements in enrichment and data acquisition may provide the proper platform to investigate the role of this perplexing PTM in health and disease.


Phototheranostics that concurrently integrates accurate diagnosis  imaging) and in situ therapy  and photothermal therapy (PTT)) into one platform represents an attractive approach for accelerating personalized and precision medicine. The second near-infrared window  has attracted considerable attention from both the scientific community and clinical doctors for improved penetration depth and excellent spatial resolution. NIR-II agents with a PDT property as well as other functions are recently emerging as a powerful tool for boosting the phototheranostic outcome. In this minireview, we summarize the recent advances of photodynamic NIR-II aggregation-induced emission luminogens (AIEgens) for biomedical applications. The molecular design strategies for tuning the electronic bandgaps and photophysical energy transformation processes are discussed. We also highlight the biomedical applications, such as image-guided therapy of both subcutaneous and orthotopic tumors, and multifunctional theranostics in combination with other treatment methods, including chemotherapy and immunotherapy; and the precise treatment of both tumor and bacterial infection. This review aims to provide guidance for PDT agents with long-wavelength emissions to improve the imaging precision and treatment efficacy. We hope it will provide a comprehensive understanding about the chemical structure–photophysical property–biomedical application relationship of NIR-II luminogens. Theranostics that concurrently integrates accurate diagnosis and in situ therapy into one platform represents an attractive approach for accelerating personalized and precision medicine ,2,3,4. LIn 2001, Tang and coworkers first coined a new photophysical phenomenon, aggregation-induced emission (AIE) , which rAnother obstacle for light-related diagnosis and therapeutics is the limited penetration depth and spatial resolution in vivo. The light-tissue interaction  is the determinant factor for optical imaging, which is highly affected by the wavelength ,58. BothIn this review, we summarize the recent advances of NIR-II AIEgens with PDT as well as other related optical properties for biomedical applications, mainly focusing on the photodynamic AIEgens with a maximal fluorescence emission wavelength above 900 nm . We will2O2), superoxide anion radical (O2•−), and hydroxyl radical (HO•). For type-II PDT, the energy of triplet excitons transfers to 3O2 to form singlet oxygen (1O2), in which the energy of the lowest triplet excited state (T1) should be higher than that of oxygen sensitization (0.98 eV) . Th. Th93]. Zhang et al., synthesized a series of AIEgens with distorted tetraphenylethylene (TPE) and diphenylamine (DPA) as donors and molecular rotors, electron-rich carbazole as a conjugated bridge, and three different positively charged fractions  as acceptors, affording three derivatives: TPEDCPy, TPEDCQu, and TPEDCAc . By incrThe cisplatin-based neoadjuvant chemotherapy (NAC), which plays an important role in combined surgical resection against microscopic and diffuse cancer cells, has become a standard of care and is supported by the latest clinical practice guidelines ,96. Howe1O2 production quantum yield of 1.05%. After injecting into HeLa tumor-bearing mice, a strong NIR-II fluorescence signal from DDTB-DP NPs was observed at the tumor site. Then, DDTB-DP NPs-mediated PTT/PDT was conducted on the residual small tumors after surgical resection; which indicated that the combination of traditional surgery and light-dependent PTT/PDT could overcome their limitations and had great clinical application potential. Finally, the NPs + PD-L1 antibodies under light were evaluated to study whether they could improve the anti-tumor immune effect of primary tumors in mice. The results showed that most tumors could be ablated without recurrence with the treatment of “NPs + light + PD-L1 antibody”. These results revealed that the combination of DDTB-DP NPs-mediated PTT/PDT and PD-L1 antibodies could achieve satisfactory tumor immunotherapy performance.Jiang et al., developed an AIEgen-based multifunctional therapeutic nanoplatform that integrated NIR-II fluorescence, photodynamic, photothermal, and immune effect . The DDTg] quinoxaline-4,9-diyl)bis(10-octyl-10H-phenothiazine) (PTZ-TQ), which had maximal absorption at 650 nm, and a significant NIR-II fluorescence emission peak at 1150 nm with a fluorescence QY of 0.3% (1O2 production quantum yield of 10% using ICG as the reference (12%). The NIR-II imaging and PDT capacities were evaluated in a nude mouse orthotopic liver tumor model. A remarkable NIR-II fluorescence signal at the liver site was recorded, which was capable of identifying the boundary between the tumor and normal liver organ. In a clinic, sometimes tumor resection cannot be performed due to the presence of many smaller tumors or a very large tumor, and multiple treatments are the common treatment methods in clinical practice. Given the strong ROS generation capacity of PTZ-TQ NPs, it was used for PDT of the residual tumor after surgery, achieving complete suppression of orthotopic tumors without recurrence.For the shallow penetration depth of most light, the applications are mainly limited to subcutaneous tumors ,101,102. of 0.3% . PTZ-TQ 2•− under 808 nm laser exposure, suggesting the type-I PDT process. The photothermal effect was further evaluated with 808 nm laser irradiation, and a high PCE of 59.6% was measured. DCTBT was assembled into liposomes by doping the EGFR-targeting peptide-modified amphiphilic polymer DSPE-PEG2000-GE11 as the encapsulation matrix to promote effective aggregation and visualization of lip-DCTBT NPs at tumor sites. In vivo NIR-II fluorescence imaging of subcutaneous PANC-1 tumor-bearing mice helped to precisely delineate the tumor site and further phototherapy exhibited significant tumor growth inhibition. More interestingly, DCTBT NPs were also able to suppress the growth of orthotopic pancreatic tumors under synergistic NIR-II fluorescence-guided type-I PDT and PTT treatments.Recently, Li et al., constructed a photosensitizer DCTBT with an AIE signature through increasing the D–A interaction and conjugation length, which possessed the functions of NIR-II fluorescence imaging, efficient type-I PDT and PTT properties . As disp1O2 generation quantum yield of 13.8% with ICG (12%) as the standard. For the excellent NIR-II fluorescence properties, as well as the good photodynamic, photothermal, and PA properties, the AIEgen was explored for imaging-guided PDT and PTT of tumors and pathogens. After 24 h post-injection of the AIE NPs into 4T1 tumor-bearing mice, the tumor region was greatly illuminated by NIR-II fluorescence and PA imaging. In contrast to the single modal imaging, this dual NIR-II/ PA imaging helped to provide complementary information about the tumor and precisely guide the subsequent phototherapy. Under the irradiation of an 808 nm laser, the tumor growth was significantly inhibited thanks to the combination of PDT and PTT. Moreover, the AIE NPs could also be used for the NIR-II fluorescence imaging of Staphylococcus aureus-infected mice, and the following imaging-guided PDT/PTT was performed to inhibit bacterial infections. Considering the close relationship between bacteria and cancers, this kind of multifunctional phototheranostic agent may represent an efficient strategy for precise diagnostics and therapeutics of bacteria-infected tumors.Recent studies have shown a close relationship between bacteria and human cancer cells, including the promotion of cancer cell development and metastasis ,106. The3)2(OTf)2 [Recently, Wang and Tang et al., reported a prismatic metal cage C-DTTP with bright NIR-II fluorescence emission through the assembly of an AIE-active four-armed ligand with a 90° Pt acceptor Pt(PEt)2(OTf)2 . As dispAs NIR-II fluorophores possess narrow electronic bandgaps, the low energy levels are usually not enough for generating ROS under light irradiation. With the endeavor of many researchers, NIR-II AIEgens with a PDT property are emerging in the last two years. The low bandgaps of NIR-II emitters result in dominated nonradiative thermal deactivation of the excited state energy in most cases. Therefore, NIR-II chromophores are naturally born with photothermal and PA properties, which enables multifunctional phototheranostics. In this minireview, we summarize the recent advances of photodynamic NIR-II AIEgens and their applications . The mol


Biomaterials have shown significant potential in various biomedical fields, including disease prevention, theranostics, and tissue regeneration by controlled delivery of therapeutics, contrast agents, sensing agents, growth factors, and so forth. In recent years, Asian scientists have contributed more to biomaterials with the increasing support of various funds. Through this Research Topic, we expected to collect several state-of-the-art advances in biomaterial science and engineering research within Asia and disseminate them to the global audience. This Research Topic comprised 35 articles  contributed by 263 authors, and it has attracted 54,023 views, 11,892 downloads, and 27 citations by researchers worldwide up to 15 January 2023. Undoubtedly, this Research Topic has won great success in displaying the significant contributions of Asian scientists to the next-generation precision medicine and further clinical translation.Luo et al.), drug delivery systems for the treatment of autoimmune diseases  and intervertebral disc degeneration (Liu and Fu), and engineering exosomes for bone defect repair . Also, anti-biofouling polymer materials with special surface wettability and their biomedical applications are summarized . Particularly, this Research Topic included two review articles about the recent development of organ-on-a-chips, regarding the application of medical imaging methods and artificial intelligence  and physiologically based pharmacokinetic (PBPK) modeling . These well-organized reviews would provide the readers with an overview and constructive perspectives of these specific topics of biomaterials.The reviews from this Research Topic have covered the latest progress in several different types of biomaterials for diverse biomedical applications, including intraocular lens materials and their surface modification for cataract surgery . Jiang et al. demonstrated that micelles containing 15-crown-5 selectively released the loaded curcumin in response to the intracellular potassium ion (K+) by forming the 2:1 ‘‘sandwich’’ host-guest complexes between 15-crown-5 and K+. Similarly, an amphiphilic micelle comprised of a Y-shaped polypeptide has demonstrated excellent drug loading and release behaviors . Interestingly, a carrier-free nanomedicine composed of ginsenosides Rg3 and Rb1 has shown better antitumor and antimetastatic effects against triple-negative breast cancer than the direct combination of free drugs . In addition, a cyclic Arg-Gly-Asp (cRGD)-decorated liposome loading with urokinase has shown enhanced thrombolytic effects by active targeting delivery of urokinase to the thrombi . Sun et al. developed electroactive shape memory polymers based on polyurethane and carbon nanotube, which would be employed as four-dimensional (4D)-printing materials for potential biomedical applications.Smart biomaterials have shown distinct and adjustable Ding et al.). Besides, copper-lithium-doped nanohydroxyapatite promoted the migration and homing of mesenchymal stem cells (MSCs) by upregulating the hypoxia-inducible factor 1α/stromal cell-derived factor-1 (HIF-1α/SDF-1) pathway . Interestingly, increasing the viscoelastic properties of cellulose nanocrystal/collagen hydrogels promoted the proliferation, alteration of shape, and matrix deposition of chondrocytes and reduced the interleukin-1β (IL-1β) secretion . Moreover, Xia et al. developed magnetic nano chains, which guided the oriented growth of neural stem cell-derived neurons. Zhang et al. demonstrated that a conductive hybrid hydrogel promoted the development of neural stem cells into neurons. Wang et al. indicated that epigallocatechin-3-gallate selenium nanoparticle scavenged reactive oxygen species (ROS) effectively and thus provided superior neuroprotective effect. Qin et al. demonstrated intimate interactions among innervation, angiogenesis, and inflammation in the condylar cartilage of temporomandibular joint osteoarthritis, which provided promising treatment targets. In addition to developing advanced biomaterials, the surface modification of materials is also a method to obtain biomaterials with specific functions. For example, coating 316L stainless steel with glycogen synthase kinase-3β inhibitor (GSKi) improved the adhesion and proliferation of human coronary artery endothelial cells . Such properties of these biomaterials made them promise to treat various diseases, such as sensory hearing loss, osteonecrosis, osteoarthritis, neurodegenerative disorders, cerebral ischemia-reperfusion injury, and cardiovascular diseases.Some biomaterials have shown capacities to manipulate the behaviors of cells. For example, graphene was proven to promote the differentiation of Lgr5+ progenitors into inner ear hair cells  (PDA-PLGA) scaffold carrying RINm5f islet cells, which supported the growth of islet cells and showed no influence on insulin secretion. When the type I diabetic rats were transplanted with the scaffold in the skeletal muscles, their blood glucose was maintained at a low level for approximately 3 weeks.In the past several decades, tissue engineering and regenerative medicine have emerged as essential branches of biomaterials. Several impressive research articles have provided constructive insights into the current Research Topic. Wang et al.) and hypoxia-pretreated MSCs  or secretomes derived from basic fibroblast growth factor (bFGF)-pretreated human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs)  have significantly facilitated the neural regeneration, which demonstrated great promise in the treatment of traumatic brain injury and the functional recovery after surgical treatment.As emerging multipotential natural biomaterials, exosomes and secretomes have exhibited bright prospects in regenerative medicine. Particularly, scaffolds containing engineered exosomes derived from adipose-derived mesenchymal stem cells (A-MSCs) under hypocapnia  induced a more uniform and dense porous morphology, greater roughness, and provided the highest shear bond strength. In addition, a multicenter retrospective study has demonstrated the high efficiency of ultrasound-guided percutaneous thermal ablation in the clinical treatment of hepatic focal nodular hyperplasia .Clinical translations and applications are the terminal goals of development of biomaterials. The unique or modified biomaterials and biotechnologies should benefit more patients only when they pass the reliability assessment and model validation. Lan et al. developed acitretin-conjugated dextran nanoparticle (ACT-Dex NP), which achieved a similar therapeutic effect on psoriasis-like skin disease at a significantly lower dosage compared to neat acitretin. This low dosage is promising to avoid acitretin’s well-known teratogenicity on fetuses. Moreover, Zhang et al. demonstrated that the neomycin-induced ototoxicity would be prevented by combination with tetrandrine  by promoting the steroid biosynthesis.Notably, the severe side effects of some medicines have limited their broad applications, although they are approved for clinical use. This Research Topic also reports strategies to reduce the side effects of clinically used therapeutics. Wang et al.). The authors reported a patient with an uncommon pseudoaneurysm, involving the left common iliac artery because of brucellosis. A favorable therapeutic effect and well prognosis were obtained with a combination treatment of long-term multi-course antibacterial therapy with combination antibiotics. This report would provide a paradigm for the clinical treatment of such diseases.This Research Topic also included a rare case report in clinic (In summary, this Research Topic encompasses a proportion of critical contributions made by Asian researchers in the biomaterial field in recent years. This regionalized Research Topic showed the rapid development of biomaterials in Asia, and it is foreseeable that Asian scientists will play a more significant role in this field in the future.


We aimed at examining both the incidence and extent of different lung perfusion abnormalities as well as the relationship between them on Tc-99m macroaggregated albumin (MAA) perfusion-only SPECT/CT scans in COVID-19 patients.Ninety-one patients  with confirmed SARS-CoV-2 virus infection were included in this retrospective study. After performing perfusion-only Tc-99m MAA SPECT/CT scans, visual, semi-quantitative assessment of the subsequent perfusion abnormalities was carried out: mismatch lesions , matched lesions , and reverse mismatch lesions . Lesion-based and patient-based analysis were performed to evaluate the extent, severity, and incidence of each perfusion abnormality. Statistical tests were applied to investigate the association between the experienced perfusion impairments.Moderately severe parenchymal lesions were detected in 87 (95.6%) patients. Although, 50 (54.95%) patients were depicted to have MM lesions, the whole patient cohort was mildly affected by this abnormality. MA lesions of average moderate severity were seen in most of the patients (89.01%). In 65 (71.43%) patients RM lesions were found with mild severity on average. Positive association was detected between total CT score and total RM score and between total CT score and total MA score. Significantly higher total CT scores were experienced in the subgroup, where RM lesions were present.Heterogeneous perfusion abnormalities were found in most of COVID-19 patients: parenchymal lesions with normal, decreased or increased perfusion and perfusion defects in healthy lung areas. These phenomena may be explained by the failure of the hypoxic pulmonary vasoconstriction mechanism and presence of pulmonary thrombosis and embolism. Increasing attention is directed towards the pandemic coronavirus disease 2019 (COVID-19) caused by SARS-CoV-2 viral infection. Viral pneumonia, featured by hypoxia, is considered to be its most common manifestation . In thiIn our Q-only SPECT/CT study heterogeneous perfusion abnormalities were found in most of the COVID-19 patients: parenchymal lesions were present with normal, decreased or increased perfusion and perfusion defects were frequently observed in healthy lung areas. These observed phenomena may be explained by the failure of the hypoxic pulmonary vasoconstriction mechanism and the presence of pulmonary thrombosis and embolism. Because our results are consistent with previous dual-energy CT and subtraction CT angiography studies, we hypothesize that Q-only SPECT/CT may be a useful tool in the evaluation of COVID-19 patients in whom CT contrast agent is contraindicated.


Monoclonal antibodies are laboratory-made proteins that mimic the immune system’s ability to fight off harmful microorganisms, including viruses such as Severe Acute Respiratory Syndrome-CoronaVirus-2 (SARS-CoV-2). The US Food and Drug Administration (FDA) and the European Medical Agency (EMA) have already authorized monoclonal antibodies of anti-SARS-CoV-2 to treat mild to moderate CoronaVIrus Disease-2019 (COVID-19) in patients at risk of developing severe disease. More recently, monoclonal antibodies anti-SARS-CoV-2 have been authorized for primary and secondary prophylaxis in patients at high risk of severe disease for background comorbidity. Primary or pre-exposure prophylaxis prevents COVID-19 in unexposed people, whereas secondary or postexposure prophylaxis prevent COVID-19 in recently exposed people to individuals with laboratory-confirmed SARS-CoV-2. This review focuses briefly on therapeutic indications of currently available monoclonal antibodies for COVID-19 pre- and postexposure prophylaxis and on the efficacy of convalescent plasma. Clostridioides difficile (CD), cytomegalovirus, hepatitis B, and are now considered promising for infectious disease targets such as newly emerging viral pathogens  [Antibodies, proteins naturally produced by the body in response to infectious stimuli, are crucial to fight infectious diseases and have been used in the prevention and treatment of bacterial and viral infections for more than a century. The history of medicine counts 5 Nobel prizes for discoveries related to treating infectious diseases with antibodies (Emil Adolf von Behring in 1901), describing humoral immunity (Paul Ehrlich and Il’ja Il’ič Mečnikov in 1908), defining the chemical structure of antibodies , the production of monoclonal antibodies (MoAbs) , and revealing the mechanism of the antibody diversity (Susumu Tonegawa in 1987). In the last 20 years, more than 60 recombinant MoAbs have been developed for human use, including rabies virus, respiratory syncytial virus (RSV), nd Zika) ,4. Theirnd Zika) . They cand Zika) . In June 2021, results from the third part of the BLAZE-2 study, a randomized, double-blind, phase 3 clinical trial designed to evaluate the efficacy of bamlanivimab MoAb in preventing COVID-19 in nursing facility nurses, staff, and residents were published . From AuMoAbs can be used even in postexposure prophylaxis to reduce the chance to get SARS-CoV-2 infection after exposition . In JuneBefore drawing final conclusions on Moab’s use, several limitations are discussed. There is a wide supply shortage in drug procurement with marked differences even in high income countries, particularly during epidemic peaks of COVID-19. Monoclonal antibodies are almost exclusively available and used in COVID-19 patients from high-income countries only, mainly due to their high cost. The time required between in vitro studies and in vivo registration trials and the regulatory approval up to the routinely clinical use are so prolonged that change trends in circulating VoCs may affect the susceptibility to currently licenced MoAb treatment . The casScientific evidence in favor of the CP use is described in patients treated with high titer plasma transfusion from 3 days to 9 days after the onset of symptoms in outpatient setting ,23. To dNew MoAbs will enter into clinical trials in the next upcoming months, and they will be evaluated for their ability to limit or modify clinical progression of SARS-CoV-2 infection. Establishing MoAb and CP therapeutic and prophylactic efficacy would represent a step forward in controlling the COVID-19 pandemic especially in the most vulnerable groups such as patients with autoimmune, oncohematologic, or neurologic diseases, particularly in resource-limited countries.


Immunotherapy is revolutionizing the clinical management of patients with different cancer types by sensitizing autologous or allogenic immune cells to the tumor microenvironment which eventually leads to tumor cell lysis without rapidly killing normal cells. Although immunotherapy has been widely demonstrated to be superior to chemotherapies, only a few populations of patients with specific cancer types respond to such treatment due to the failure of systemic immune activation. In addition, severe immune-related adverse events are rapidly observed when patients with very few responses are given higher doses of such therapies. Recent advances of lipid-based nanoparticles (NPs) development have made it possible to deliver not only small molecules but also mRNAs to achieve systemic anticancer immunity through cytotoxic immune cell activation, checkpoint blockade, and chimeric antigen receptor cell therapies, etc. This review summarized recent development and applications of LNPs in anticancer immunotherapy. The diversity of lipid-based NPs would encapsulate payloads with different structures and molecular weights to achieve optimal antitumor immunity through multiple mechanisms of action. The discussion about the components of lipid-based NPs and their immunologic payloads in this review hopefully shed more light on the future direction of anticancer immunotherapy. The first anticancer immunotherapeutic agent was used T cell growth factor interleukin 2 (IL-2), where IL-2 treatment showed significant tumor suppression by enhancing T-cell activation in patients with metastatic kidney cancer and melanoma , and loss of bioactivity after long-term administration have limited the immunostimulatory efficacy of small molecules therapeutics . UnfortuLipid-based NPs exhibit various types of structure. Majority of the lipid-based NPs exhibited near spherical shape with one or more lipid external layers Figure 2in vivo delivery purposes. 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), a biodegradable analogue of DOTMA, was then developed for lipid-based NPs gene delivery (1/2) of pDNA was extended to 7 h in mice compared to naked pDNA, which had a t1/2 of less than 10 min. The prolonged circulation time of lipid/pDNA particles in mice resulted in dose accumulation in distal tumors over 24 h with minor accumulation in liver and spleen  and low transfection potency for gene delivery , 25. Thedelivery . Dimethydelivery . The firdelivery . The autd spleen . With thd spleen –34.Ionizable lipids were a series of lipids with native pKa values below 7. It was designed to enhance anionic drug encapsulation and minimize systemic toxicities by cationic lipids due to permanent positive charges. Lipid-based NPs embedded with ionizable lipids could load anionic payloads at pH values below the pKa of ionizable lipids when it is positively charged. Then the drug-loaded lipid-based NPs are suitable for systemic delivery with a neutral exterior depending on physiological pH values . When trin vivo through i.v. administration  and 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane (DODMA) were firstly developed for antisense oligonucleotides (ASOs) delivery with significantly increased circulation half-lives of ASOs stration . Later, stration . In anotstration . Recent stration . MC3 exh005mg/kg . Under t005mg/kg –54.II) phase  phase .Lipid bilayers formed by helper lipids can be stabilized by cholesterol where cholesterol can fill the geometry gaps between phospholipids . The incin vitro or in vivo. Nonetheless, preclinical immunogenicity studies are recommended when developing nanomedicines containing fusogenic oils.Fusogenic oils are special components in NEs which is made of fatty acids, lipid substitutes, waxes, oil-soluble vitamins, and other lipophilic components. The viscosity, refractive index, density, phase behavior, and interfacial tension of the oils greatly determine the stability and functional properties of NEs . Other fin vivo than LNPs containing PEG2000-DSG, though PEG2000-DSG could prolong circulation times for LNPs in blood compared to PEG2000-DMG  or macrophages) serve to early detect cancers by sensing tumor-associated antigens or pattern-/damage-associated molecular patterns (PAMPs/DAMPs) through pattern recognition receptors (PRRs) . Activat. Antitum. Poly  can induce specific gene regulation which became a new therapeutic area in infectious diseases, neurodegenerative disorders, cancer, and other rare diseases. Besides direct targeting of specific oncogenes, RNAi could also potentiate antitumor immune responses by downregulating immune-suppressive proteins Figure 3in vitro , suggestin vitro . Since Fin vitro . Transfoin vitro , and siRin vitro . When usin vitro . Direct in vitro .miRNAs also indirectly facilitate the interaction between tumors and host immune systems which can be targeted for either downregulation or replacement by RNAi. Oncogenic miRNAs have been reported to promote tumor progression by inducing M2-like TAM in TMEs  and tumor-specific antigens (TSAs). Neoantigens expressed in the mutated tumor microenvironment would allow the development of personalized cancer vaccines with patient-specific neoepitopes. Tumor antigens against cancer mainly depend on the delivery of tumor antigen peptides or coding mRNAs , 135. Dein vivo  . The autThe safety, immunogenicity, and tolerability of the first personalized IVAC MUTANOME , a poly-neoepitope-coding RNA vaccine, have been evaluated in phase I clinical trials (NCT02035956) targeting mutant neoantigens for the treatment of patients with melanoma. A strong immune response against the vaccine antigens was observed. T cell response was also generated against 60% of the 125 selected neoepitopes with no adverse drug reactions, indicating good tolerability of the vaccine by enrolled patients . Many otIn a phase I study (NCT02410733), FixVac, a complex of RNA/lipoplexes against malignant melanoma TAAs New York-ESO 1, tyrosinase, melanoma-associated antigen A3 (MAGE-A3), and trans-membrane phosphatase with tensin homology (TPTE) showed the metabolic activity of the spleen increased post the sixth immunization, indicating the targeted delivery of an mRNA-LNP encoding four non-mutated melanoma antigens and activation of resident immune cells . Within in vitro and in vivo , CAR-T cell therapy, and NK cell therapy. Despite their tremendous potential, adoptive cell therapies also raised concerns regarding the unwanted immunological side effects and insertional mutagenesis in the human genome due to the use of viral vectors for ineering , 164. Inineering , 164. Th therapy –167. Lip therapy . mRNA/litoxicity . Therefotoxicity Figure 4toxicity . Soon, Bed CAR-T . Lipid-bed CAR-T . TRAIL-ein vivo . Anotherin vivo . CAR-eng release . Lipid-b release .The main efforts of lipid-based NPs in clinical trials against cancer are to deliver chemotherapies and therapeutic nucleic acids as discussed in previous sections. However, the number of lipid-based NPs formulations successfully approved in the market is still limited, suggesting that the development of lipid-based NPs remains certain barrier in the translation from preclinical animal models to humans.Non-specific gene regulation by lipid-based NPs may take place when lipid-based NPs are used for gene delivery. It is noticeable that although the structure of lipid-based NPs is almost similar to those of lipid nanoparticles or micelles expressed by cells, the synthesized functional lipids, such as cationic or ionizable lipids, are still considered exogenous lipid-like materials which may be potentially recognized by PRRs on the cell surface. The high amount of exogenous functional lipids within lipid-based NPs may further trigger unwanted cell cycle arrest, cellular metabolism, or immune responses, which would disrupt the targeted gene regulation by nucleic acid payloads in lipid-based NPs , 176. Whin-situ therapeutic efficacy (Although lipid-based NPs are believed to passively target and accumulate in tumors through the enhanced permeability and retention (EPR) effect, a growing body of research has revealed that lipid-based NPs, without any ligand-modification for active targeting, mainly accumulate in the liver upon systemic administration . Such efefficacy . Therefoin vivo to achieve the enormous antitumor activity but also are capable of ex vivo engineering cancer cell therapies with comparable efficiency compared to other non-viral or viral vectors. However, the formulation and large-scale manufacturing process of lipid-based NPs need to be optimized from industrial aspects. Depending on the specific drug payloads and applications, different helper lipids and cationic/ionizable lipids need to be selected to address specific challenges to immunotherapeutic delivery. For small-molecule chemotherapies or ligands for surface PPRs, lipid-based NPs with high colloidal stability should be designed to prolong their systemic retention time. Therefore, lipids with higher Tm are preferred due to their long-term stability in storage and the systemic fluid. For RNAi, mRNA, and ligands targeting intracellular PPRs, lipid-based NPs should be designed to satisfy membrane fusion and endosomal escape, such as embedding PE-based lipids and decreasing PEG-lipids, where the stability for lipid-based NPs may be compromised. Fortunately, the challenges for colloidal stability and in vivo side effects by lipid-based NPs may be bypassed in the application of cell therapies where lipid-based NPs containing mRNAs can be formulated and transfected into T/NK cells ex vivo in a real-time manner. However, due to the transient gene transfection by lipid-based NPs/mRNA, the durability of tumor-killing activities by engineered T/NK cells need to be further evaluated thoroughly. In addition, there are stilling remaining concerns for the delivery of lipid-based NPs, such as immunogenicity and tissue accumulation. Therefore, efforts should also be made on developing lipid-based NPs targeting specific organs or cell-populations as well as utilizing biodegradable formulation components with low immunogenicity (Lipid-based NPs represent the most advanced and widely applicable delivery vehicles for small molecules and nucleic acids. In cancer immunotherapy, lipid-based NPs not only could deliver small molecules and mRNA therapies genicity , 187.Moreover, the therapeutic payloads need to be designed for optimal lipid-based NP delivery efficiency and antitumor immunity. Small molecule drugs can be designed with ionizable structures to be stably encapsulated into the lipid-based NPs, and their mechanism of actions should be well identified without off-target effects such as inducing immunosuppression or autoreactive immune disorders . NucleicConceptualization, ZZ; writing—original draft preparation, ZZ and SY; writing—review and editing, ZZ, SY, YH, XZ, and RL. All authors have read and agreed to the published version of the manuscript.https://biorender.com/.All figures are created with YH and XZ are employed by The Whiteoak Group, Inc.The remaining author declares 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.


Given emerging evidence of immune escape in the SARS-CoV-2 Omicron viral variant, and its dominance, effectiveness of heterologous and homologous boosting schedules commonly used in low-to-middle income countries needs to be re-evaluated. We conducted a test-negative design using consolidated national administrative data in Malaysia to compare the effectiveness of homologous and heterologous BNT162b2, CoronaVac, and AZD1222 booster vaccination against SARS-CoV-2 infection in predominant-Delta and predominant-Omicron periods. Across both periods, homologous CoronaVac and AZD1222 boosting demonstrated lower effectiveness than heterologous boosting for CoronaVac and AZD1222 primary vaccination recipients and homologous BNT162b2 boosting. Broadly, marginal effectiveness was smaller by 40–50 percentage points in the Omicron period than the Delta period. Without effective and accessible second-generation vaccines, heterologous boosting using BNT162b2 for inactivated and vectored primary vaccination recipients is preferred. Immunogenic and real-world studies documented immune escape in Omicron amongst inactivated and mRNA primary and booster vaccine recipients . MalaysiTo address this issue, we used a test-negative design derived from the vaccinated adult (aged ≥ 18) population from national administrative data: (i) register of COVID-19 vaccine recipients, (ii) register of supervised and approved reverse transcription polymerase chain reaction and antigen rapid tests conducted at all testing facilities, (iii) the COVID-19 confirmed cases line listing, (iv) register of healthcare “frontliners”, and (v) “contacts”  identified by Malaysia's automated contact tracing system. Vaccination status is defined as “primary vaccinated” and “boosted” at 14 days post-dose two and three, respectively, at the point of testing.Absent of adequate genomic surveillance, the Bai-Perron sequential structural break test  was applAll analyses were executed in Python 3.9, R 4.1.2, and EViews 11. Significance level of 5% was used for statistical inference.The Bai-Perron sequential breakpoint test estimated 5 February 2022 as the start of the predominant-Omicron period. The study population comprises individuals aged 18 and above without prior infections who were boosted between 27 October 2021 and 4 February 2022, or primary vaccinated between 1 July 2021 and 30 September 2021. These correspond to 0–3 months post-boosted, and 4–6 months post-primary vaccinated, respectively. The latter reflects that effectiveness wanes 3 months post-primary vaccination [This study has three strengths. Firstly, to the best of our knowledge, this is the first comparison of homologous and heterologous booster effectiveness for CoronaVac and AZD1222 primary vaccination recipients under Delta and Omicron dominance in a low-to-middle income country (LMIC). Secondly, this study considers a diverse set of commonly used homologous and heterologous vaccinations. Thirdly, granular national administrative data underpinned the analysis.The main limitation is the potential undermeasurement of SARS-CoV-2 infections due to the use of self-test kits. Nevertheless, supervised test rates were higher during the Delta and Omicron periods  than the rest of 2021 . While Malaysia adopted a symptomatic testing strategy, asymptomatic SARS-CoV-2 infections may be included as there were no formal barriers to access supervised testing. Due to the rapid rollout of boosters, and short follow-up duration, potential waning of effectiveness could not be analysed. Importantly, while a wide range of confounders were adjusted for, including exposure risk, health-seeking behaviour, underlying conditions, and batch effects, residual confounding may still be present in observational studies such as this.First-generation boosters were less effective against Omicron than Delta SARS-CoV-2 infection. In both periods, heterologous boosting was more effective than homologous boosting for AZD1222 and CoronaVac primary vaccination recipients. Our analysis complements pre-Omicron findings that homologous boosting with AZD1222 in the United Kingdom  and CoroThis is part of The Real-World Evaluation of COVID-19 Vaccines under the Malaysia National COVID-19 Immunization Programme (RECoVaM) study registered in the National Medical Research Register (NMRR-21-1660-60697). This study was granted ethical approval by the Medical Research and Ethics Committee (MREC), Ministry of Health, Malaysia.Click here for additional data file.


A 69-year old woman presented to Dermatology for a skin cancer screening. Clinical examination found a 4-mm skin-colored papule on her forehead. The lesion had been present for 3 years, was asymptomatic, and had remained unchanged .Fig 1A wDermatoscopy revealed a well-circumscribed homogeneous brown-orange color papule with linear vessels at the periphery of the lesion that were out-of-focus  in contrReflectance confocal microscopy (RCM) of the reticular dermis showed a central area with rounded dermal nodules of different sizes with a white dot pattern, surrounded by bright collagen bundles, with no plump-bright cells outside the tumor area. Palisading and peripheral dark, cleft-like spaces, which are typical diagnostic features of basal cell carcinoma, were absent , Fig 6.FHistopathology showed photodamaged skin with sharply demarcated basophilic nodules in the dermis, composed of basaloid cells with a small hyperchromatic nucleus and scant cytoplasm and some larger pale cells with an ovoid nucleus. The cells were surrounding globules of eosinophilic basement-membrane material. Moreover, duct-like structures and some cystic cavities with eosinophilic material could be identified  and 8.Fi,,Spiradenomas are uncommon, well-differentiated, benign, dermal neoplasms originating in sweat glands. They typically appear as small solitary nodules, except when they are found in Brooke-Spiegler syndrome, in which case they are multiple and occur with other adnexal neoplasias.None disclosed.


Patients with hematologic diseases receiving anti-CD20 monoclonal antibodies show a low seroconversion rate after two doses of mRNA Coronavirus disease-2019 (COVID-19) vaccines, especially within 12 months of their last anti-CD20 antibodies . PreviouThis prospective study investigated humoral and cellular responses and safety after a third dose (booster) of the mRNA COVID-19 vaccines in patients with hematologic diseases at Kobe City Medical Center General Hospital. Patients were recruited between January and July 2022. In this trial, adult patients with non-Hodgkin B-cell lymphoma, idiopathic thrombocytopenic purpura (ITP), or acquired thrombotic thrombocytopenic purpura (TTP) were eligible if they received their last dose of rituximab or obinutuzumab within 12 months before two doses of SARS-CoV-2 vaccine. We excluded patients with a known and undocumented history of SARS-CoV-2 infection and comorbidities of autoimmune diseases, except ITP and TTP. We also included adult healthy volunteers without a history of COVID-19 to serve as the control group. The institutional review board approved this study and informed consent was obtained from all patients and volunteers. The blood serum of the study participants was collected before and 14–90 days after the third vaccination. We extracted relevant data from the medical records. All participants answered a questionnaire asking about local or systemic adverse events within seven days after the third vaccination. The primary endpoint was set as the proportion of subjects who acquired anti-S1 IgG antibodies. We also evaluated anti-S1 IgG titers, T-cell responses against SARS-CoV-2, and adverse events after vaccination as secondary endpoints. The details of the data, the kit used for antibody detection and T-cell responses, and statistical issues are described in the supplemental data.Sixty-three patients with hematologic diseases who received anti-CD20 antibodies and 30 healthy controls were enrolled in the study. Fifty-three patients and all the healthy controls were included in the final analysis. Ten patients were excluded from the final analysis: four patients were infected with SARS-CoV-2 before the completion of blood sampling, two patients had undocumented COVID-19 infection, two did not have blood sampling before the third vaccination, and two did not receive their third dose. Blood samples have collected a median of ten  days before the third vaccination and a median of 228 (IQR 202–245) days after the second vaccination. The characteristics of patients and healthy volunteers are presented in Table Humoral responses were evident in 8 (15%) and 22 (42%) patients before and after the third vaccination, respectively. All healthy volunteers remained seropositive before and after their third doses Fig. . T-cell Humoral responses are critical for blocking the acquisition of viral infection . In thisIn addition, it should be noted that the anti-S1 antibody titers in patients were substantially lower than those in the healthy volunteers. Anti-S1 IgG titers have been reported to be associated with neutralization activity against SARS-CoV-2, and an observational study has indicated that neutralization activity is absent if anti-S1 antibody levels are low . Among pCellular immune responses most likely control viral replication after infection and prevent progression to severe diseases . The preThis study has several limitations. First, the sample size was too small to establish the predictors of impaired humoral and cellular responses. Second, the regimens used in combination with the anti-CD20 antibodies were heterogeneous. Therefore, the potential effect of medications other than anti-CD20 antibodies on humoral and cellular responses should be considered. Third, all participants received monovalent COVID-19 vaccines for their third dose. Impaired neutralizing responses induced by the third monovalent COVID-19 vaccines have been reported against the current variants in circulation, such as BQ.1 and XBB .In summary, we confirmed that the third vaccination with an mRNA-based COVID-19 vaccine induces humoral and cellular immune responses in patients who recently received anti-CD20 antibodies. In addition, some seronegative patients achieved seroconversion after the third vaccination. A T-cell response against SARS-CoV-2 was identified after the third vaccination in some patients who failed to gain humoral response, indicating the benefit of prompt, up-to-date vaccination among the vulnerable population. We need more studies to determine the factors associated with impaired vaccine effectiveness and the optimal timing of vaccination in B cell-depleted patients.Supplementary Data


In-person directly observed therapy (DOT) is commonly used for tuberculosis (TB) treatment monitoring in the US, with increasing usage of video-DOT (vDOT). We evaluated the impact of COVID-19 on TB treatment adherence, and utilization and effectiveness of vDOT.We abstracted routinely collected data on individuals treated for TB disease in Baltimore, Maryland between April 2019 and April 2021. Our primary outcomes were to assess vDOT utilization and treatment adherence, defined as the proportion of prescribed doses (7 days/week) verified by observation (in-person versus video-DOT), comparing individuals in the pre- and post-COVID (April 2020) periods.Among 52 individuals with TB disease, 24 (46%) received treatment during the COVID-19 pandemic. vDOT utilization significantly increased post-COVID (18/24[75%]) compared to pre-COVID . Overall, median verified adherence was similar pre- and post-COVID . Adherence was significantly higher overall when using vDOT (median 86% [IQR 70-98%]) compared to DOT ; this improved adherence with vDOT was evident in both the pre-COVID  and post-COVID period .vDOT utilization increased post-COVID and was more effective than in-person DOT at verifying ingestion of prescribed treatment. Tuberculosis (TB) remains a public health priority in the United States (US).–3 TB treDirectly observed therapy (DOT) is a strategy used to monitor TB treatment adherence. While a Video DOT (vDOT) is a modality where digital technology platforms  are used to verify the observation of a patient, either live or recorded, receiving their TB treatment., 10 WHO In early 2020, the onset of the COVID-19 pandemic created a need to continue providing essential healthcare services while adopting social distancing practices to reduce the speed of viral transmission. This spurred a rapid transition towards telehealth clinical services beginning in March and April 2020 across health systems. Since thWe conducted a retrospective observational cohort study of patients receiving routine TB treatment at the BCHD TB Program. The study protocol was approved by the Johns Hopkins Medicine Institutional Review Board.From April 2019 to April 2021, we enrolled patients receiving treatment for TB disease in Baltimore, Maryland. Routinely collected data was abstracted from eligible patients ≥ 18 years of age who were then classified into the pre-COVID period and post-COVID period. Individuals were classified as receiving treatment in the post-COVID period if they started therapy after April 2020, or received at least three months of therapy after April 2020 to capture those who may have initiated prior to April 2020 but received significant care during the post-COVID period. As part of routine clinical care at BCHD, all patients have treatment adherence monitored using DOT (in-person adherence verification through observation in person’s home or workplace 5 days per week excluding weekends and government holidays) or asynchronous vDOT (7 days per week) according to local protocols using a shared decision-making paradigm between patients and TB clinic providers. Patients and providers changed treatment monitoring strategies as needed based on individualized considerations; there were no restrictions or exclusions on vDOT usage based on prior adherence or treatment completion, sputum smear-status, or drug resistance.The vDOT system  is comprised of a smartphone/tablet application (app) used by patients, and a web-based dashboard used by the TB clinic. The patient-side app reminds patients to take their medications on a schedule specified by the clinician. For non-English speaking patients, the app can deliver content in multiple languages customized to the patient. The app allows a secure/encrypted video recording to be taken as the patient ingests the prescribed medications, according to the patient’s schedule. Encrypted videos and data are transmitted to a secure cloud-based server within seconds and subsequently deleted from the patient’s phone, ensuring privacy. The videos can then be asynchronously viewed to verify medication adherence by TB clinic staff through the emocha web interface, typically on the next business day, where the clinician is prompted to make a determination as to adherence with DOT after watching each video.Primary outcomes were stratified by the pre- and post-COVID periods and included vDOT utilization and treatment adherence. vDOT utilization was defined as the proportion of patients who received any vDOT for treatment of TB disease. Treatment adherence was defined as the proportion of verified prescribed doses over 7 days per week. Standard descriptive statistics were used to characterize the study cohort. Chi-square test and two-sample t-tests were used to compare vDOT utilization and treatment adherence between DOT and vDOT overall and in the pre- and post-COVID periods. Multivariable logistic regression was used to understand the relationship between patient characteristics with utilization of vDOT and treatment adherence. All analyses were performed with Stata 16.A total of 52 patients received treatment for active TB during the study period, of which 24 (46%) received therapy in the post-COVID period. Most patients were men , Black/African American , and born outside the US . Nearly all patients  were HIV-uninfected. Thirty patients were classified as having pulmonary TB (58%), 17 (33%) patients had extrapulmonary TB, and 5 (10%) patients had both. There was no difference in the distribution of pulmonary and extrapulmonary TB (p = 0.81) or AFB smear-status (p = 0.29) in the pre- and post-COVID periods. TB treatment was monitored for all patients with vDOT, in-person DOT, or both. Overall vDOT was utilized for some portion of TB treatment monitoring for 58% (30/52) of patients while the remaining patients  received treatment monitoring with in-person DOT only. Among 30 individuals receiving vDOT, 14 (47%) received vDOT for their entire treatment course within the BCHD TB program. A significantly greater proportion of patients utilized vDOT during the post-COVID period , compared to the pre-COVID period .  Among inMedian overall adherence (irrespective of monitoring strategy) was 66% (IQR 57–84%), and was similar in the pre-  and post-COVID  periods (p = 0.96). Overall, the proportion of prescribed doses  verified through observation was significantly higher when utilizing vDOT (median 86% [IQR 70%-98%]), compared to in-person DOT . These findings were driven by the high proportion of self-administered doses when using in-person DOT (median 38% [IQR 35–44%) compared to periods when patients were using vDOT (median 1% [IQR 0–7%]) (p < 0.01). Verified adherence was higher when using vDOT in both the pre-COVID  and post-COVID period . When comparing the pre- and post-COVID period adherence for each treatment monitoring strategy, verified adherence was similar when using in-person DOT  and declined slightly when using vDOT . The latter difference was primarily attributable to an increase in the number of doses that were self-administered when using vDOT between the pre- and post-COVID periods . Self-administered doses  when using in-person DOT remained high and stable across the pre-and post-COVID periods .  Overall,The COVID-19 pandemic and associated lockdowns and staffing shifts led to significant disruption in healthcare delivery within TB programs including the Baltimore City Health Department, particularly related to traditional approaches to treatment verification using in-person DOT., 15 WithOverall, there have been shifting paradigms in TB treatment monitoring towards patient-centered approaches such as vDOT. Our study suggests that the COVID-19 pandemic may have accelerated these transitions. In Baltimore, prior to COVID-19, less than half of all patients were monitored using vDOT. With limitations on in-person contact during much of 2020 and 2021, we found that nearly three-quarters of patients were subsequently monitored using vDOT. Age greater than 65 was associated with decreased vDOT utilization, a finding previously noted in the vDOT literature, which may suggest that familiarity with smartphone/tablet technology may impact patient acceptance of this strategy. Despite Our study therefore highlights particular limitations with in-person DOT in monitoring TB treatment, which is increasingly prescribed 7 days per week. We found that vDOT was more effective than DOT at verifying treatment adherence, and that less than two-thirds of prescribed therapy were able to be verified when using in-person DOT–reflecting logistical challenges with in-person verification during weekends and holidays. Consequently, we report that a large proportion of prescribed therapy is given with self-administration when using in-person DOT.We also identified other challenges experienced by TB programs during the COVID-19 pandemic., 17 SpecThere are several limitations with this study. Since the study is an analysis of the real-world implementation of vDOT in a city health department, patients were not randomized to one modality or another. Although we report significant differences of verified adherence between vDOT and in-person DOT, the study was not designed to draw a definitive conclusion in this regard for all populations. Selection of patients for vDOT versus in-person DOT were made by clinic staff based on individualized considerations, and reflect routine clinical practices. While vDOT was the utilized by a significant majority of patients in the post-COVID period, additional qualitative assessments are needed to fully understand patient and provider perspectives associated with this transition from in-person DOT. Overall, vDOT had been available in the BCHD TB program since approximately 2016. Nonetheless, vDOT utilization increases appeared to be influenced by the COVID-19 pandemic. However, we cannot rule out alternative factors including increasing comfort and experience with the vDOT platform that may also have led to progressive increases in vDOT utilization over time.In conclusion, despite the significant disruption of healthcare activities during the COVID-19 pandemic, verified TB treatment adherence at the BCHD was sustained in the setting of increased vDOT utilization. Verified TB treatment was significantly higher with vDOT and adds another data point to the growing literature showing that vDOT improves upon in-person DOT for adherence monitoring and is readily accepted by patients., 18, 19


Testicular germ cell tumors are the most common malignancy in young and middle-aged men. Spontaneous primary testicular tumor regression, or testicular tumor burn-out, is a rare clinical phenomenon where extragonadal metastatic lesions are observed concurrently with the spontaneous regression of the primary testicular germ cell tumors. Here, we describe the case of a 36-year-old male who presented to our hospital with left-sided abdominal pain and testicular swelling and was found to have significant retroperitoneal lymphadenopathy on his abdominopelvic CT scan. His testicular ultrasound showed multiple echogenic calcifications through the right testicle consistent with microlithiasis. Biopsy of the retroperitoneal lesion revealed a mixed germ cell tumor of testicular origin composed of embryonal carcinoma and teratoma. The patient received four cycles of bleomycin, etoposide, and cisplatin, followed by retroperitoneal lymph node dissection (RPLND) and radical right testicular orchiectomy. Here, we report the second case of burned-out testicular tumor in a patient with ipsilateral cryptorchidism. Furthermore, we elucidate the etiology, clinical presentation, and diagnostic modalities in burned-out testicular germ cell tumors. Spontaneous primary tumor regression, or tumor burn-out, refers to the clinical phenomenon where the metastatic disease is present without any evidence of the primary tumor . The burA 36-year-old male with a history of right testicular cryptorchidism status post orchiopexy at the age of four presented to the hospital complaining of worsening left lower pelvic and groin pain for the past two months. He also had concurrent right testicular swelling, night sweats, and an unintentional 15 lbs weight loss. The patient’s physical exam revealed diffuse abdominal distention, mild left costovertebral angle tenderness, and a significantly enlarged right testicle. His abdominopelvic CT revealed diffuse retroperitoneal lymphadenopathy, the largest conglomeration measuring 7.8 cm x 6.5 cm, encasing the left ureter, causing moderate obstructive hydronephrosis Figure , while tThe patient’s serum tumor markers revealed a significantly elevated β-human chorionic gonadotropin, lactate dehydrogenase, and α-fetoprotein. He had a core needle biopsy of the left retroperitoneal lymph node, which showed small fragments of neoplastic tissue admixed with skeletal muscle, with myxoid stroma and multinucleated syncytiotrophoblasts consistent with embryonal carcinoma. Benign glandular elements were also present, suggesting a component of teratoma. The results were consistent with mixed GCT of testicular origin Figure .The patient’s positron emission tomography (PET) revealed increased metabolic activity in the mediastinal and retroperitoneal lymph nodes. He was diagnosed with stage 3C (cTxN3S2 M1a) mixed non-seminoma GCT. He received four cycles of neoadjuvant chemotherapy consisting of bleomycin 30 mg, etoposide 100 mg, and cisplatin 20 mg (BEP) once every three weeks for four cycles, without any treatment-related complications. The patient’s tumor markers were normalized post-chemotherapy, although his PET scan showed persistent para-aortic and retroperitoneal lymphadenopathy. The patient underwent retroperitoneal lymph-node dissection (RPLND), removing 14 para-aortic and paracaval lymph nodes, followed by radical right inguinal orchiectomy. The microscopic examination of paracaval lymph nodes revealed necrotic fibro-adipose tissue with hemorrhage but no neoplastic cells, and the dissected peri-aortic lymph nodes revealed one node containing mature teratoma while the remaining seven showed no neoplastic tissue. The histopathologic exam of the testicle revealed calcifications and scar tissue, confirming burned-out GCTs. The patient’s post-op PET scan showed no residual malignancy, and he has remained in remission until now, 11 months post-operation.Burned-out testicular tumors are disseminated neoplastic lesions of testicular origin without an active malignancy in the testes . Prym etWhile the pathogenesis of spontaneous testicular tumor regression remains elusive, three leading theories have emerged: 1) T-cell mediated response against the primary testicular GCT occurs only after the extragonadal tumor dissemination, which allows for the cytotoxic T-cells to recognize and mount an immune response against the neoplastic cells; 2) primary testicular tumor ischemia caused by vascular supply and demand mismatch due to its high metabolic demands lead to primary tumor regression; 3) the de-novo development of neoplastic testicular germ cells occur in the extragonadal tissue -11. In tA distinguishing feature of this case was the patient’s history of right testicle cryptorchidism, which was subsequently the primary tumor location. Although cryptorchidism is a well-known risk factor for the subsequent development of testicular cancer, to our best knowledge, only one prior study has reported a burned-out testicular tumor in a patient with cryptorchidism .The diagnosis of burned-out testicular GCTs is challenging as it has no distinct clinical, laboratory, or imaging characteristics -14. WhenSpontaneous primary tumor regression, or tumor burn-out, is a rare clinical phenomenon where there are metastatic tumor cells but with partial or complete disappearance of the primary tumor in the tissue of origin. Here, we reported a 36-year-old male with a history of ipsilateral right testicular cryptorchidism status post orchiopexy who presented with retroperitoneal lymphadenopathy diagnosed as a mixed germ-cell tumor of testicular origin composed of embryonal carcinoma and teratoma. However, his testicular ultrasound revealed no lesions suspicious of malignancy. The patient received four cycles of bleomycin, etoposide, and cisplatin, followed by radical right inguinal orchiectomy and RPLND. The microscopic examination of the right testicle revealed multiple microcalcifications and scar tissues confirming a burned-out testicular germ cell tumor. Meanwhile only one of the 14 dissected lymph nodes showed mature teratoma. The patient’s post-op imaging showed no residual malignancy, and he has remained in remission until now. This report describes the etiology, clinical presentation, and diagnostic modalities in burned-out testicular germ cell tumors. Furthermore, it examines the theories on the pathogenesis of testicular tumor burn-out. To our best knowledge, this is the second reported case of burned-out testicular tumor in a patient with ipsilateral cryptorchidism.