Chimeric Structure Research Articles

How Much Do We Know About the Mouse Respiratory Epithelial Stem Cells?

Background-Purpose of this study: Tissue-specific stem cell lines are useful tools for cell biology studies. Information on respiratory tissue cell lines is limited. A doxycycline-regulated epithelial precursor cell line was established from the lung tissue of a tTAxSV40 Tag double transgenic mouse. In this study, we have characterized this cell line in vitro & in vivo, and found to mimic a rare subpopulation of club- and pneumocyte type II-dual cells. Methods: It was partially characterized using cell viability and death assays, H3-thymidine incorporation assay, chloride efflux assay, Western blotting of proteins secreted, RT-PCR assays for RNA isolated. In addition, immune-deficient SCID mice were used as hosts for implantation of this precursor cell line, and feed with/without doxycycline containing water. Immunofluorescent typing using different antibodies were used to characterize the implanted lung. Results: This cell line was found to mimic a rare subpopulation of club- and pneumocyte type II- dual cells with multiple phenotypes. Cell growth was doxycycline-regulated and observed only when doxycycline was omitted from the medium or present at concentrations up to 1 µg/ml, higher concentrations were inhibitory. ACT+ ciliated cells were found upon implantation into immune-deficient mice, in addition. Cell growth was doxycycline-regulated in vitro. When transplanted subcutaneously into immune-deficient mice, these cells migrated to the lung to form organized chimeric structures of donor and host origins, with club cells in the terminal bronchioles, ACT+ ciliated cells along the epithelial lining, and pneumocyte type II-cells in the alveolar interstices. No such homing of donor cells to the lung was observed when the implanted mice were fed doxycycline-containing water. Discussions-Conclusions: This lung stem cell line might be able to provide us with an insight into the differentiation pathway of lung epithelial cells as well as with some understanding of the nature of air trophic-pulmonary epithelial cells. The results of this study underline the possibility of a future application for somatic (stem / precursor) cells in tissue replacement and tissue engineering of the damaged lung. Its ability to secrete and deliver soluble protein, might be a potential novel way for drug delivery. In addition, stem cells are thought to proliferate and differentiate in response to a deficiency or as a result of injury. Successful migration to the target organ and subsequent maturation of these precursors could be attributed to a requirement of lung stem cells to search for an aerated environment. Our findings challenge some current concepts of stem cell biology.This lung stem cell line may become a rich source of cells for tissue engineering and cell-based therapy for lung injury. The route and protocol established for cell introduction into the lung may provide a novel alternative to delivery of soluble protein substances through the airways. This lung stem cell line might also be modified to provide models for screening drugs against respiratory infection.

Study on the mechanism and reaction characteristics of red-mud-catalyzed pyrolysis of corn stover

With catalytic pyrolysis gradually becoming the most efficient thermal treatment for biomass, the search for catalysts with high economic and catalytic performances has attracted widespread attention from scholars. In this study, the use of red mud as a catalyst for the pyrolysis of biomass to produce high-quality pyrolysis gas was investigated. The effects of the three components of red mud (Fe2O3, Al2O3, and CaCO3) on pyrolysis and the catalytic effect and mechanism of red mud on pyrolysis were analyzed. Fe2O3 was the most dominant catalytically active ingredient in red mud, which had a good contribution to the improvement of carbon conversion and hydrogen yield. Specifically, Fe2O3-catalyzed corn stover pyrolysis at a 40 % addition ratio increased the hydrogen yield by 82 %. Because of the stable pore structure provided by Al2O3 and SiO2 as carriers and the chimeric structure of the catalyst particles, the specific surface area of the red mud reached 25.88 m2/g, and the specific surface area of the magnetic separation product reached 42.41 m2/g, which greatly improved the catalytic performance. Red-mud-catalyzed corn stover pyrolysis with a red mud addition ratio of 40 wt% could achieve an H2 yield of 108 ml/g, representing a rise of 79 %. The phase composition and crystal structure of the red mud and magnetic separation products were analyzed; the components in the red-mud catalyst doped with each other at the lattice level in the form of a solid solution, which improved the catalytic activity.

Characteristics of genetic changes in the <i>SMN1</i> gene in spinal muscular atrophy 5q

Background. Proximal spinal muscular atrophy 5q (5q‑SMA) is one of the most common neuromuscular diseases, which is caused by mutations of the SMN1 gene. Despite the fact that most studies consider SMN1 “deletion” as the most common cause of 5q‑SMA, gene loss is in fact associated with both classical deletions and conversions of SMN1 and SMN2, as well as with formation of chimeric structures. Up to now, far too little attention has been paid to the prevalence of types of SMN1 loss. However, different types of mutations can have different influence on the clinical findings and the effectiveness of therapy. A deeper study of the structure of these genes will allow us to determine the predictors of response to treatment and bring us closer to understanding the reasons for the instability of the SMN region.Aim. To study genetic changes in the SMN1 gene, as well as the number of copies of the SMN1 and SMN2 genes in 5q‑SMA.Materials and methods. The study involved 703 patients, for whom the analysis of the number of copies of SMN1 and SMN2 was performed in the center of molecular medicine of I.P. Pavlov First Saint Petersburg State Medical University for 2018–2021. Gene copy number analysis was performed by multiplex ligation‑dependent probe amplification (MLPA) using the SALSA MLPA P021 SMA kit (MRC Holland).Results. Among 703 participants, a homozygous SMN1 deletion was found in 167 (24 %), 76 (11 %) were carriers of the deletion and no aberrations were found in the remaining 460 (65 %). Among patients with a homozygous deletion, 41 (24 %) cases of a true deletion were identified. Also, 11 (7 %) cases of partial deletion with a homozygous loss of the 7th exon and heterozygous of the 8th exon of SMN1 were identified. The most common type of aberration was the conversion of SMN1 to SMN2 – 94 (56 %) cases, which is characterized by a homozygous loss of SMN1 and a reciprocal increase in SMN2 copies. In addition, 21 (13 %) cases of the formation of hybrid genes were found.Conclusion. Types of genetic aberrations in 5q‑SMA have not been studied enough yet. A search of the literature revealed very few studies which results are comparable with ours. However, further research of genetic changes in SMN1 and SMN2 might bring more clarity on the causes and mechanisms of this disease, and get us closer to finding the most effective method of treatment.

Reverse vaccinology approach to identify novel and immunogenic targets against Porphyromonas gingivalis: An in silico study.

Porphyromonas gingivalis is a primary causative agent of chronic periodontitis. Moreover, it leads to several systemic diseases, including rheumatoid arthritis, cardiovascular, neurodegenerative, and Alzheimer’s diseases. It seems that the development of a vaccine against this bacterium is necessary. Thus, this study decided to identify novel immunogenic targets and developed multiple epitope-based vaccines against P. gingivalis. For this purpose, the pan/core-proteome of this bacterium was studied, and the suitable vaccine targets were selected based on different properties, including exposed localization of proteins, antigenicity, non-allergenicity, non-similarity to host proteome, stability, B-cell epitopes and MHC II binding sites, sequence conservation, molecular docking, and immune simulation. Through the quartile scoring method, 12 proteins with ≥ 20 scores were considered as suitable immunogenic targets. The results of the protein domain and functional class search showed that most of the immunogenic proteins were involved in the transport and metabolism of inorganic ions and lipids. In addition, two unknown function proteins, including WP_004584259.1 and WP_099780539.1 were detected as immunogenic targets. Three constructions carrying multi-epitopes were generated including Naked, LCL, and as chimeric structures. Among them, FliC chimeric protein had the strongest affinity to the human TLR2, 4, and 6, while the LCL platform represented the highest level of immune stimulation response. The obtained results from this study revealed new insights into prophylactic routes against P. gingivalis by introducing novel immunogenic targets. However, further investigations, including site-directed mutation and immunoassay are needed to confirm the pathogenic role and protectivity of these novel proteins.

Comparison of Mitochondrial Genomes between a Cytoplasmic Male-Sterile Line and Its Restorer Line for Identifying Candidate CMS Genes in Gossypium hirsutum.

As the core of heterosis utilization, cytoplasmic male sterility (CMS) has been widely used in hybrid seed production. Previous studies have shown that CMS is always closely related to the altered programming of mitochondrial genes. To explore candidate CMS genes in cotton (Gossypium hirsutum), sequencing and de novo assembly were performed on the mitochondrial genome of the G. hirsutum CMS line SI3A, with G. harknessii CMS-D2 cytoplasm, and the corresponding G. hirsutum restorer line 0-613-2R. Remarkable variations in genome structure and gene transcripts were detected. The mitochondrial genome of SI3A has three circle molecules, including one main circle and two sub-circles, while 0-613-2R only has one. RNA-seq and RT-qPCR analysis proved that orf606a and orf109a, which have a chimeric structure and transmembrane domain, were highly expressed in abortive anthers of SI3A. In addition, comparative analysis of RNA-seq and full-length transcripts revealed the complex I gene nad4 to be expressed at a lower level in SI3A than in its restorer and that it featured an intron retention splicing pattern. These two novel chimeric ORFs and nad4 are potential candidates that confer CMS character in SI3A. This study provides new insight into the molecular basis of the nuclear–cytoplasmic interaction mechanism, and that putative CMS genes might be important sources for future precise design cross-breeding of cotton.

Multiepitope-based vaccine design by exploring antigenic potential among leptospiral lipoproteins using comprehensive immunoinformatics and structure-based approaches.

Leptospirosis is a tropical and globally neglected zoonotic disease caused by pathogenic spirochetes, Leptospira. Although the disease has been studied for decades, a potent or effective vaccine is not available so far. Efforts are being made to design an efficient vaccine candidate using different approaches. Immunoinformatics approaches have been proven to be promising in terms of time and cost. Here, we used immunoinformatics and structure-based approaches to evaluate antigenic B- and T-cell epitopes present on the leptospiral lipoproteins (LipL). The promiscuous overlapping epitopes (B-cell, T-cell, interferon (IFN)-γ positive, and non-allergens), which can induce humoral, cell-mediated, and innate immunity, were selected to generate a multiepitope chimeric vaccine. To enhance the vaccine immunogenicity, a Toll-like receptor (TLR) agonist was fused to the vaccine with a suitable linker. The chimeric vaccine structure was predicted for molecular docking studies with immune receptors. Moreover, the stability of the vaccine-immune receptor complexes was analyzed by normal mode analysis (NMA). The potency of the vaccine construct was predicted by the immune simulation tool. The study provides additional information toward constructing peptide-based chimeric vaccines against Leptospira.

In silico prediction and in vitro validation of the effect of pH on adhesive behaviour of the fused CsgA-MFP3 protein

Recombinant production of mussel foot proteins among marine-inspired proteinaceous adhesive materials has been attracted high attention for medical applications, due to their exceptional versatility potential of hierarchically arranged nanostructures. Various biochemical and proteinous factors such as amyloid CsgA curli protein have been used as a synergistic factor to enhance the constancy of obtained bio-adhesion but their mechanistic interactions have not yet been deeply investigated widely in different pH conditions. To this end, the present study has first sought to assess molecular simulation and prediction by using RosettaFold to predict the 3-dimensional structure of the fused CsgA subunit and the MFP3 protein followed by in vitro verification. It was developed an ensemble of quantitative structure-activity relationship models relying on simulations according to the surface area and molecular weight values of the fused proteins in acidic to basic situations using PlayMolecule (protein preparation app for MD simulations) online databases followed by molecular dynamic simulation at different pHs. It was found that acidic conditions positively affect adhesive strength throughout the chimeric structure based on comparative structure-based analyses along with those obtained in prevailing literature. Atomic force microscopy analysis was confirmed obtained in silico data which showed enhanced adhesive properties of fused protein after self-assembly in low pH conditions. In conclusion, the augmented model for reactivity predictions not only unravels the performance and explain ability of the adhesive proteins but in turn paves the way for the decision-making process for chimeric subunits modifications needed for future industrial production.

Ectomycorrhizal Networks in the Anthropocene: From Natural Ecosystems to Urban Planning.

Trees acquire hydric and mineral soil resources through root mutualistic associations. In most boreal, temperate and Mediterranean forests, these functions are realized by a chimeric structure called ectomycorrhizae. Ectomycorrhizal (ECM) fungi are highly diversified and vary widely in their specificity toward plant hosts. Reciprocally, association patterns of ECM plants range from highly specialist to generalist. As a consequence, ECM symbiosis creates interaction networks, which also mediate plant–plant nutrient interactions among different individuals and drive plant community dynamics. Our knowledge of ECM networks essentially relies on a corpus acquired in temperate ecosystems, whereas the below-ground facets of both anthropogenic ECM forests and inter-tropical forests remain poorly investigated. Here, we successively (1) review the current knowledge of ECM networks, (2) examine the content of early literature produced in ECM cultivated forests, (3) analyze the recent progress that has been made in understanding the place of ECM networks in urban soils, and (4) provide directions for future research based on the identification of knowledge gaps. From the examined corpus of knowledge, we reach three main conclusions. First, the emergence of metabarcoding tools has propelled a resurgence of interest in applying network theory to ECM symbiosis. These methods revealed an unexpected interconnection between mutualistic plants with arbuscular mycorrhizal (AM) herbaceous plants, embedding ECM mycelia through root-endophytic interactions. This affinity of ECM fungi to bind VA and ECM plants, raises questions on the nature of the associated functions. Second, despite the central place of ECM trees in cultivated forests, little attention has been paid to these man-made landscapes and in-depth research on this topic is lacking. Third, we report a lag in applying the ECM network theory to urban soils, despite management initiatives striving to interconnect motile organisms through ecological corridors, and the highly challenging task of interconnecting fixed organisms in urban greenspaces is discussed. In particular, we observe a pauperized nature of resident ECM inoculum and a spatial conflict between belowground human pipelines and ECM networks. Finally, we identify the main directions of future research to make the needed link between the current picture of plant functioning and the understanding of belowground ECM networks.

Detection of recombinant breakpoint in the genome of human enterovirus E11 strain associated with a fatal nosocomial outbreak

BackgroundThe aim of this study was to characterize the genome of a recombinant Enterovirus associated with severe and fatal nosocomial infection; it was typed as Echovirus 11 (E-11) according to the VP1 gene. Enterovirus infection is generally asymptomatic and self-limited, but occasionally it may progress to a more severe clinical manifestation, as in the case described here. Recombination plays a crucial role in the evolution of Enteroviruses (EVs) and has been recognized as the main driving force behind the emergence of epidemic strains associated with severe infection. Therefore, it is of utmost importance to monitor the circulation of recombinant strains for surveillance purposes.MethodsEnterovirus-RNA was detected in the serum and liver biopsy of patients involved in the nosocomial cluster by commercial One-Step qRT-PCR method and the Enterovirus strains were isolated in vitro. The EVs typing was determined by analyzing the partial-length of the 5′UTR and VP1 sequences with the web-based open-access Enterovirus Genotyping Tool Version 0.1. The amplicons targeting 5′UTR, VP1 and overlapping fragments of the entire genome were sequenced with the Sanger method. Phylogenetic analysis was performed comparing the VP1 and the full-genome sequences of our strains against an appropriate reference set of Enterovirus prototypes of the Picornaviridae genera and species retrieved from the Enterovirus Genotyping Tool. Recombination analysis was performed using RDP4 software.ResultsThe Neighbor-Joining tree of the VP1 gene revealed that the 4 patients were infected with an identical molecular variant of Echovirus 11 (E-11). While the phylogenetic and the RDP4 analysis of the full-genome sequences provided evidence that it was a chimeric strain between an E-11 and a Coxsackievirus B (CV-B).ConclusionsThe chimeric structure of the E-11 genome might have contributed to the severe infection and epidemic feature of the strain, but further biological characterizations are needed. The evidence reported in this study, highlights the limit of typing techniques based on the VP1 gene, as they fail to identify the emergence of recombinant strains with potentially more pathogenic or epidemic properties, thus providing only partial information on the epidemiology and pathogenesis of Enteroviruses.

Factors That Influence the Choice of Markov Model Order in Discriminating DNA Sequences from Different Sources.

Markov models have frequently been used in genetic sequence analysis. The number of parameters of a Markov model increases exponentially with model order, so it is often recommended that the order be chosen based on the size of data being modeled, lower orders for small and higher orders for large dataset sizes. Approaches based on model selection criterion have also been proposed. An important problem in microbiology and evolutionary biology is to decipher chimeric genomes of microbes, particularly, identify segments of distinct ancestries in genomes and reconstruct the plausible evolutionary scenarios that might have shaped the chimeric genomes in the microbial world. In this study, we assessed a Markov model-based segmentation method for its ability to detect compositionally disparate segments in chimeric sequence constructs as a function of model order, sequence length, and phylogenetic divergence. Our results show that the choice of Markov model order depends on both sequence size and composition. Higher order Markov models were found to be more effective in delineating sequence segments arising from closely related organisms in longer constructs; on the other hand, lower order Markov models were found to be more appropriate in delineating sequence segments arising from distantly related organisms in shorter constructs. These findings are important and timely, with broad implications in fields such as epidemiology that has to deal with the emergence of novel pathogenic chimeras that arise by foreign DNA acquisition, and ecology where chimeric structures may arise in various ecosystems, necessitating more robust approaches for their deconstruction and interpretation.

Influenza chimeric hemagglutinin structures in complex with broadly protective antibodies to the stem and trimer interface

Influenza virus hemagglutinin (HA) has been the primary target for influenza vaccine development. Broadly protective antibodies targeting conserved regions of the HA unlock the possibility of generating universal influenza immunity. Two group 2 influenza A chimeric HAs, cH4/3 and cH15/3, were previously designed to elicit antibodies to the conserved HA stem. Here, we show by X-ray crystallography and negative-stain electron microscopy that a broadly protective antistem antibody can stably bind to cH4/3 and cH15/3 HAs, thereby validating their potential as universal vaccine immunogens. Furthermore, flexibility was observed in the head domain of the chimeric HA structures, suggesting that antibodies could also potentially interact with the head interface epitope. Our structural and binding studies demonstrated that a broadly protective antihead trimeric interface antibody could indeed target the more open head domain of the cH15/3 HA trimer. Thus, in addition to inducing broadly protective antibodies against the conserved HA stem, chimeric HAs may also be able to elicit antibodies against the conserved trimer interface in the HA head domain, thereby increasing the vaccine efficacy.

Structure-Activity Relationship Investigations of Novel Constrained Chimeric Peptidomimetics of SOCS3 Protein Targeting JAK2.

SOCS3 (suppressor of cytokine signaling 3) protein suppresses cytokine-induced inflammation and its deletion in neurons or immune cells increases the pathological growth of blood vessels. Recently, we designed several SOCS3 peptidomimetics by assuming as template structures the interfacing regions of the ternary complex constituted by SOCS3, JAK2 (Janus Kinase 2) and gp130 (glycoprotein 130) proteins. A chimeric peptide named KIRCONG chim, including non-contiguous regions demonstrated able to bind to JAK2 and anti-inflammatory and antioxidant properties in VSMCs (vascular smooth muscle cells). With the aim to improve drug-like features of KIRCONG, herein we reported novel cyclic analogues bearing different linkages. In detail, in two of them hydrocarbon cycles of different lengths were inserted at positions i/i+5 and i/i+7 to improve helical conformations of mimetics. Structural features of cyclic compounds were investigated by CD (Circular Dichroism) and NMR (Nuclear Magnetic Resonance) spectroscopies while their ability to bind to catalytic domain of JAK2 was assessed through MST (MicroScale Thermophoresis) assay as well as their stability in biological serum. Overall data indicate a crucial role exerted by the length and the position of the cycle within the chimeric structure and could pave the way to the miniaturization of SOCS3 protein for therapeutic aims.

Chimeric Structures in Mental Illnesses-"Magic" Molecules Specified for Complex Disorders.

Mental health problems cover a wide spectrum of diseases, including mild to moderate anxiety, depression, alcohol/drug use disorders, as well as bipolar disorder and schizophrenia. Pharmacological treatment seems to be one of the most effective opportunities to recover function efficiently and satisfactorily. However, such disorders are complex as several target points are involved. This results in a necessity to combine different types of drugs to obtain the necessary therapeutic goals. There is a need to develop safer and more effective drugs. Considering that mental illnesses share multifactorial processes, the paradigm of one treatment with multiple modes of action rather than single-target strategies would be more effective for successful therapies. Therefore, hybrid molecules that combine two pharmacophores in one entity show promise, as they possess the desired therapeutic index with a small off-target risk. This review aims to provide information on chimeric structures designed for mental disorder therapy (i.e., schizophrenia and depression), and new types of drug candidates currently being tested. In addition, a discussion on some benefits and limitations of multifunctional, bivalent drug candidates is also given.

CTX-M-15-producing Klebsiella pneumoniae ST273 associated with nasal infection in a domestic cat

The aim of this study was to investigate the genetic context of expanded-spectrum β-lactam resistance in a Klebsiella pneumoniae strain causing a hard-to-treat nasal infection in a domestic cat. A K. pneumoniae isolate was recovered from a 4-year-old male cat hospitalised in a veterinary hospital in Paraíba, Northeastern Brazil. Following phenotypic confirmation of multidrug resistance by the disk diffusion method, the genome was sequenced using an Illumina MiSeq system. Multilocus sequence typing (MLST) and structural features related to antimicrobial resistance were determined by downstream bioinformatics analyses. The strain was confirmed as sequence type 273 (ST273) K. pneumoniae harbouring a variety of genes conferring antimicrobial resistance to phenicols tetracyclines, aminoglycosides, β-lactams, fosfomycin, sulfonamides and quinolones. Two plasmids were identified. Plasmid p114PB_I co-harboured a set of plasmid-borne resistance genes [blaCTX-M-15, blaTEM-1, qnrS1, tetD, tetR, sul2, aph(6)-Id, aph(3'') and cat2]. Notably, the multiresistance region was characterised as a chimeric plasmid structure sharing high sequence homology with several plasmids from Enterobacteriaceae. The second plasmid (p114PB_II) was characterised as a plasmid present in many genomes belonging to K. pneumoniae. The genetic context of the plasmid sequences harboured by a veterinary pathogenic K. pneumoniae isolate reveals the high complexity of horizontal gene transfer mechanisms in the acquisition of antimicrobial resistance genes. The emergence, dissemination and evolution of antimicrobial resistance must be investigated from a One Health perspective.

Chimeric Antigen Receptor Structure and Manufacturing of Clinical Grade CAR Engineered Cells using Different Bioreactors.

Increasing success of adaptive cell therapy (ACT), such as genetically engineered T cells to express chimeric antigen receptors (CARs) proven to be highly significant technological advancements and impressive clinical outcomes in selected haematological malignancies, with promising efficacy. The evolution of CAR designs beyond the conventional structures is necessary to address some of the limitations of conventional CAR therapy and to expand the use of CAR T cells to a wider range of malignancies. There are various obstacles with a wide range of engineering strategies in order to improve the safety, efficacy and applicability of this therapeutic modality. Here we describe details of modular CAR structure with all the necessary domains and what is known about proximal CAR signalling in T cells. Furthermore, the global need for adoptive cell therapy is expanding very rapidly, and there is an urgent increasing demand for fully automated manufacturing methods that can produce large scale clinical grade high quality CAR engineered immune cells. Despite the advances in automation for the production of clinical grade CAR engineered cells, the manufacturing process is costly, consistent and involves multiple steps, including selection, activation, transduction, and Ex-Vivo expansion. Among these complex manufacturing phases, the choice of culture system to generate a high number of functional cells needs to be evaluated and optimized. Here we list the most advance fully automated to semi-automated bioreactor platforms can be used for the production of clinical grade CAR engineered cells for clinical trials but are far from being standardized. New processing options are available and a systematic effort seeking automation, standardization and the increase of production scale, would certainly help to bring the costs down and ultimately democratise this personalized therapy. In this review, we describe in detail different CAR engineered T cell platforms available and can be used in future for clinical-grade CAR engineered ATMP production.

Galectin-3 facilitates inflammation and apoptosis in chondrocytes through upregulation of the TLR-4-mediated oxidative stress pathway in TC28a2 human chondrocyte cells.

Osteoarthritis (OA) is a common degenerative joint disease. The pathological changes of chondrocytes involve oxidative stress, the pro-inflammatory response, and pro-apoptotic events. Galectin-3 (Gal-3) is a 35 kDa protein with a special chimeric structure. Gal-3 participates in the progression of many diseases, such as cancer metastasis and heart failure. A previous study demonstrated that Gal-3 expression in human cartilage with OA is increased. However, the role of Gal-3 in chondrocyte dysfunction in joints is still unclear. In this study, we applied Gal-3 (5-20 μg/ml) to TC28a2 human chondrocyte cells for 24 h to induce chondrocyte dysfunction. We found that Gal-3 upregulated TLR-4 and MyD88 expression and NADPH oxidase, thereby increasing intracellular ROS in the chondrocytes. Gal-3 increased phosphorylated MEK1/2 and ERK levels, and promoted NF-κB activity. This activation of NF-κB was reduced by silencing TLR-4 and NOX-2. In addition, Gal-3 caused apoptosis of chondrocytes through the mitochondrial-dependent pathway via the TLR-4/NADPH oxidase/MAPK axis. Our study proves the pathogenic role of Gal-3 in Gal-3-induced chondrocyte dysfunction and injuries.

Amelioration of acute myocardial infarction injury through targeted ferritin nanocages loaded with an ALKBH5 inhibitor

The roles of m6A RNA methylation and mitochondrial metabolism in acute myocardial infarction (AMI) remain unclear. In this study, we demonstrated that m6A RNA methylation affected ischemia/reperfusion (I/R) injury in AMI through the “Erasers” protein ALKBH5-related metabolic reprogramming, characterized by the inhibition of enzyme activities of the tricarboxylic acid cycle; moreover, a surface-modified bioengineered ferritin nanocage was obtained from Archaeoglobus fulgidus, with a chimeric structure containing 8 lysine residues, SpyTag/SpyCatcher, and the C1q ligand Scarf1, which could disassemble and self-assemble in neutral solutions according to different Mg2+ concentrations. The surface-modified bioengineered ferritin nanocage targeted the dying cells in the infarct area under the guidance of Scarf1. These cells were then phagocytosed through recognition of their TfR1 receptor. Lysosomal escape was achieved through the 8 lysine residues on the nanocage, and the nanocage disassembled based on the differences in intracellular and extracellular Mg2+ concentrations. Finally, the ALKBH5 inhibitor IOX1 was loaded onto the ferritin nanocage and used in the AMI model, and it was found to effectively improve cardiac function. These results provide a potential strategy for the treatment of AMI in the future. Statement of significanceIn acute myocardial infarction (AMI) induced by ischemia/reperfusion injury, m6A RNA methylation aggravates the injury through the “Erasers” protein ALKBH5-related metabolic reprogramming. To achieve precise treatment, genetic engineering-based recombinant expression technology was used to obtain a ferritin from Archaeoglobus fulgidus. The obtained ferritin was designated as HAfFtO, and it can disassemble and self-assemble in a neutral solution under different Mg2+ concentrations and achieve lysosomal escape. Three G4S linkers were used to connect SpyTag with HAfFtO to synthesize HAfFtO-ST and recombination Scarf1 containing SpyCatcher structure, namely SC-Sf. According to the SpyTag/SpyCatcher technique, HAfFtO-ST and SC-Sf can form a gentle and firm combination, namely HSSS. The ALKBH5 inhibitor IOX1 was loaded on HSSS to form HSSS-I. HSSS-I effectively improved the cardiac function and decreased the infarct size in AMI.

Viral Environmental Warfare: Technology Bill of Rights Critically Needed

The ongoing “hidden in plain sight” geoengineering operations including the pervasive, near global spraying of ultrafine particles into the troposphere pose an existential risk to the biosphere and humanity. Likewise, bioengineering and genetic manipulation of potential pandemic pathogens with associated accidental or deliberate release represents a dire risk to the modern world. Compelling evidence to date indicates that the chimeric structure and added furin cleavage site of SARS-CoV-2 was the result of human manipulation in the laboratory. Coronavirus and vaccine patents indicate a foreknowledge of the COVID-19 pandemic, which conveniently serves the interests of the global elites. The new COVID-19 vaccines, hailed as the answer to the pandemic, have potential toxicity, fail to prevent transmission, drive the development of new variants, and over time may predispose recipients to antibody-dependent enhancement (ADE) of infection. Other important technological threats include Internet-based censorship, nano-particulate pollution, electro-pollution and the massive amounts of electromagnetic energy inflicted on humans and Earth’s natural environment. A Technology Bill of Rights is critically needed to protect humanity and salvage what remains of earth’s life support systems.

Chimerism of Carbon by Ruthenium Induces Gradient Catalysis

AbstractImproving the round‐trip efficiency and cycle life of lithium‐oxygen batteries (LOBs) through the cathode design is essential for their employment in electric vehicles. Here, a neoteric chimeric air‐cathode is proposed, in which ultra‐dispersed ruthenium (Ru) nanoparticles are partially confined by microtube walls of free‐standing carbon textiles, but petty parts of Ru with catalytic surfaces are exposed. The special chimeric structure benefits not only for preventing nanocatalysts agglomeration but also tailoring the formation/decomposition mechanisms of lithium peroxide (Li2O2) during oxygen reduction/evolution reactions. Critically, theoretical simulations disclose that the enlarged chimeric catalyst‐support interphase between the Ru and carbon matrix enhances the electron density of Ru particles and then induces a gradient Gibbs free energy of LiO2 adsorption energy at active sites, and thus fundamentally modulates the morphology evolution mechanism of finial Li2O2. In the LOBs, the chimeric electrode affords a remarkably high energy efficiency of 84.1% and long‐term cyclability (260 cycles at 300 mA g−1).

Exploring SARS-COV-2 structural proteins to design a multi-epitope vaccine using immunoinformatics approach: An in silico study

In December 2019, a new virus called SARS-CoV-2 was reported in China and quickly spread to other parts of the world. The development of SARS-COV-2 vaccines has recently received much attention from numerous researchers. The present study aims to design an effective multi-epitope vaccine against SARS-COV-2 using the reverse vaccinology method. In this regard, structural proteins from SARS-COV-2, including the spike (S), envelope (E), membrane (M), and nucleocapsid (N) proteins, were selected as target antigens for epitope prediction. A total of five helper T lymphocytes (HTL) and five cytotoxic T lymphocytes (CTL) epitopes were selected after screening the predicted epitopes for antigenicity, allergenicity, and toxicity. Subsequently, the selected HTL and CTL epitopes were fused via flexible linkers. Next, the cholera toxin B-subunit (CTxB) as an adjuvant was linked to the N-terminal of the chimeric structure. The proposed vaccine was analyzed for the properties of physicochemical, antigenicity, and allergenicity. The 3D model of the vaccine construct was predicted and docked with the Toll-like receptor 4 (TLR4). The molecular dynamics (MD) simulation was performed to evaluate the stable interactions between the vaccine construct and TLR4. The immune simulation was also conducted to explore the immune responses induced by the vaccine. Finally, in silico cloning of the vaccine construct into the pET-28 (+) vector was conducted. The results obtained from all bioinformatics analysis stages were satisfactory; however, in vitro and in vivo tests are essential to validate these results.