Target Function Research Articles

Criterion of construction and strategy of functioning of information and telecommunication system with protection against destructive effects

The creation of information and telecommunication systems with the integration of information transmission, reception and processing functions determines the need to find technical solutions to ensure its security and protection while increasing the intensity of information processes, improving technologies for unauthorized access to information areas and creating destructive effects. Joint fulfillment of requirements for information security and maintaining operational properties of the system, which regulate efficiency, efficiency and resource intensity of information exchange, is achieved in accordance with a minimum-margin rule, the essence of which is to find the maximum possible information losses and determine a strategy for achieving minimum values of these losses. Within the framework of the used approach, the best solution is to establish a set of parameters of transmitting and receiving devices and information processing means with the lowest loss value of the target function of information exchange under the most dangerous and effective destructive effects. Goal – justification of the criterion for construction and strategy of functioning of the information and telecommunication system with protection against destructive effects. The criterion of construction and strategy of operation of the information and telecommunication system are justified in order to achieve the best probabilistic-time indicators of information exchange with a balanced set of measures to ensure security and information protection in case of destructive impacts. Due to the variety and variability of the conditions for the manifestation of destabilizing factors, the parameters and reconfiguration schemes of telecommunication channels, as well as the operating modes of information processing devices, were established while achieving the effectiveness of meeting the information needs of subscribers according to the optimality criterion and setting indicators of the efficiency and resource consumption of the system according to the suitability criterion. Possibilities and options for adaptive, situational and reflective control of the information and telecommunication system in the interests of rational distribution of the information resource to achieve target functions and the use of technologies for ensuring security and protecting information from destructive influences are investigated. Automatic control of the system is at the stage of prohibiting destructive effects on the information area by maintaining components in states that most fully correspond to the goals and objectives of information exchange. Situational control of information security during transmission and reception, used, as a rule, at an early stage of implementation of the conflict component in the information process, is based on the identification of destabilizing factors based on the results of classification of current states and neutralization of threats due to the restructuring of the system operation regulations carried out. Reflexive control, the strategy of which is based on prediction and evaluation of changes in the parameters of information processes under destructive effects, is in the form of transferring components of the information and telecommunication system to the state of its own information equilibrium, which helps to maximize the target function of information exchange while minimizing the performance indicators of destructive effects carried out. Due to the expediency of consistent implementation of the above control strategies in assessing the degree of occurrence of threats to the information process, the vector of parameters of information objects and telecommunication channels must from the conditions for neutralizing the full set of predicted destabilizing factors be determined. A reasonable criterion for building an information and telecommunication system makes it possible to jointly search for technical solutions for creating receiving and transmitting devices and information processing means in order to achieve maximum performance indicators of information exchange under established limitations of efficiency and resource intensity. The developed rule for selecting the strategy for controlling components of the information and telecommunication system establishes time-frequency and space-energy distributions of information processes necessary to meet the information needs of subscribers when transmitting, receiving and processing information in different conditions for the manifestation of destructive threats. The obtained results form a methodological basis for building information and telecommunication systems with protection of information coming through their channels with increasing requirements for target functions of information exchange and strengthening system restrictions due to improvement of technologies for unauthorized access to information areas and creation of destructive effects.

Technical Systems Effectiveness Management Technique

Currently, the task of performing a target function by a technical system with minimal resource costs is urgent. Obtaining the beneficial effect of a vehicle depends on many factors, which can become the basis for the formation of a control effect on the functional part of the system. At the same time, it is necessary to take into account the importance of a useful (target) effect in order to manage the technical systems effectiveness. The purpose of this article is to describe an instrument that allows adaptively redistributing the resource spent on the implementation of a purposeful process in the vehicle, based on the requirements of the consumer function and the target effect. As a result of the conducted research, the article proposes a technique for redistributing the resource of a technical system in the interests of its effectiveness management. The features of the presented technique are the visibility of its results presentation, as well as the possibility of application in evaluating the effectiveness and it`s management in relation to various types of technical systems, which will allow to substantiate the practical significance of obtained results.

Targeted photodynamic therapy technique of Janus nanoparticles on breast cancer

Spherical gold/polyacrylic acid (Au/PAA) polymer–inorganic Janus nanoparticles (JNPs) with simultaneous therapeutic and targeting functions were fabricated. The obtained Au/PAA JNPs were further selectively functionalized with folic acid (FA) and thiol PEG amine (SH-PEG-NH2) on Au sides to provide superior biocompatibility and active targeting, while the other PAA sides were loaded with 5-aminolevulinic acid (5-ALA) to serve as a photosensitizer (PS) for photodynamic therapeutic (PDT) effects on MCF-7 cancer cells. The PS loading of 5-ALA was found to be 83% with an average hydrodynamic size and z-potential of 146 ± 0.8 nm and −6.40 mV respectively for FA-Au/PAA-ALA JNPs. The in vitro PDT study of the JNPs on MCF-7 breast cancer cells under 636 nm laser irradiation indicated the cell viability of 24.7% ± 0.5 for FA-Au/PAA-ALA JNPs at the IC50 value of 0.125 mM. In this regard, the actively targeted FA-Au/PAA-ALA JNPs treatment holds great potential for tumour therapy with high cancer cell-killing efficacy.

Formulation optimization of lyophilized aptamer-gold nanoparticles: Maintained colloidal stability and cellular uptake

Anti-nucleolin (NCL) aptamer AS1411 is the first anticancer aptamer tested in clinical trials. Gold nanoparticles (AuNP) have been widely exploited for various biomedical applications due to their unique functional properties. In this study, we evaluated the colloidal stability and targeting capacity of AS1411-funtionalized AuNP (AuNP/NCL-Apt) against MCF-7 breast cancer cell line before and after lyophilization. Trehalose, mannitol, and sucrose at various concentrations were evaluated to determine their cryoprotection effects. Our results indicate that sucrose at 10 % (w/v) exhibits the best cryoprotection effect and minimal AuNP/NCL-Apt aggregation as confirmed by UV–Vis spectroscopy and dynamic light scattering (DLS) measurements. Moreover, the lyophilized AuNP/NCL-Apt at optimized formulation maintained its targeting and cytotoxic functionality against MCF-7 cells as proven by the cellular uptake assays utilizing flow cytometry and confocal laser scanning microscopy (CLSM). Quantitative PCR (qPCR) analysis of nucleolin-target gene expression also confirmed the effectiveness of AuNP/NCL-Apt. This study highlights the importance of selecting the proper type and concentration of cryoprotectant in the typical nanoparticle lyophilization process and contributes to our understanding of the physical and biological properties of functionalized nanoparticles upon lyophilization.

Multifaceted roles of lymphatic and blood endothelial cells in the tumor microenvironment of hepatocellular carcinoma: A comprehensive review.

The tumor microenvironment is a complex network of cells, extracellular matrix, and signaling molecules that plays a critical role in tumor progression and metastasis. Lymphatic and blood vessels are major routes for solid tumor metastasis and essential parts of tumor drainage conduits. However, recent studies have shown that lymphatic endothelial cells (LECs) and blood endothelial cells (BECs) also play multifaceted roles in the tumor microenvironment beyond their structural functions, particularly in hepatocellular carcinoma (HCC). This comprehensive review summarizes the diverse roles played by LECs and BECs in HCC, including their involvement in angiogenesis, immune modulation, lymphangiogenesis, and metastasis. By providing a detailed account of the complex interplay between LECs, BECs, and tumor cells, this review aims to shed light on future research directions regarding the immune regulatory function of LECs and potential therapeutic targets for HCC.

Polymer-based antibody mimetics (iBodies) target human PD-L1 and function as a potent immune checkpoint blocker

Immune checkpoint blockade (ICB) using monoclonal antibodies against programmed cell death protein 1 (PD-1) or programmed death-ligand 1 (PD-L1) is the treatment of choice for cancer immunotherapy. However, low tissue permeability, immunogenicity, immune-related adverse effects, and high cost could be possibly improved using alternative approaches. On the other hand, synthetic low-molecular-weight (LMW) PD-1/PD-L1 blockers have failed to progress beyond in vitro studies, mostly due to low binding affinity or poor pharmacological characteristics resulting from their limited solubility and/or stability.Here, we report the development of polymer-based anti-human PD-L1 antibody mimetics (α-hPD-L1 iBodies) by attaching the macrocyclic peptide WL12 to a N-(2-hydroxypropyl)methacrylamide copolymer. We characterized the binding properties of iBodies using surface plasmon resonance, enzyme-linked immunosorbent assay, flow cytometry, confocal microscopy, and a cellular ICB model. We found that the α-hPD-L1 iBodies specifically target human PD-L1 (hPD-L1) and block the PD-1/PD-L1 interaction in vitro, comparable to the atezolizumab, durvalumab, and avelumab licensed monoclonal antibodies targeting PD-L1. Our findings suggest that iBodies can be used as experimental tools to target hPD-L1 and could serve as a platform to potentiate the therapeutic effect of hPD-L1-targeting small molecules by improving their affinity and pharmacokinetic properties.

Synthesizing Amino Acids Modified with Reactive Carbonyls in Silico to Assess Structural Effects Using Molecular Dynamics Simulations.

Protein carbonylation by reactive aldehydes derived from lipid peroxidation leads to cross-linking, oligomerization, and aggregation of proteins, causing intracellular damage, impaired cell functions, and, ultimately, cell death. It has been described in aging and several age-related chronic conditions. However, the basis of structural changes related to the loss of function in protein targets is still not well understood. Hence, a route to the in silico construction of new parameters for amino acids carbonylated with reactive carbonyl species derived from fatty acid oxidation is described. The Michael adducts for Cys, His, and Lys with 4-hydroxy-2-nonenal (HNE), 4-hydroxy-2-hexenal (HHE), and a furan ring form for 4-Oxo-2-nonenal (ONE), were built, while malondialdehyde (MDA) was directly attached to each residue. The protocol describes details for the construction, geometry optimization, assignment of charges, missing bonds, angles, dihedral angles parameters, and its validation for each modified residue structure. As a result, structural effects induced by the carbonylation with these lipid derivatives have been measured by molecular dynamics simulations on different protein systems such as the thioredoxin enzyme, bovine serum albumin and the membrane Zu-5-ankyrin domain employing root-mean-square deviation (RMSD), root mean square fluctuation (RMSF), structural secondary prediction (DSSP) and the solvent-accessible surface area analysis (SASA), among others.

Recent Progress in DNA Damage Response-Targeting PROTAC Degraders.

DNA damage response (DDR) defects in cells play a crucial role in tumor development by promoting DNA mutations. These mutations create vulnerabilities specific to cancer cells, which can be effectively targeted through synthetic lethality-based therapies. To date, numerous small molecule DDR inhibitors have been identified, and some of them have already been approved for clinical use. However, due to the complexity of the tumor microenvironment, mutations may occur in the amino acid residues of DDR targets. These mutations can affect the efficacy of small molecule inhibitors targeting DDR pathways. Therefore, researchers have turned their attention to next-generation DNA damage repair modulators, particularly those based on PROTAC technology. From this perspective, we overviewed the recent progress on DDR-targeting PROTAC degraders for cancer therapy. In addition, we also summarized the biological functions of different DDR targets. Finally, the challenges and future directions for DDR-target PROTAC degraders are also discussed in detail.

The ‘PSILAUT’ protocol: an experimental medicine study of autistic differences in the function of brain serotonin targets of psilocybin

BackgroundThe underlying neurobiology of the complex autism phenotype remains obscure, although accumulating evidence implicates the serotonin system and especially the 5HT2A receptor. However, previous research has largely relied upon association or correlation studies to link differences in serotonin targets to autism. To directly establish that serotonergic signalling is involved in a candidate brain function our approach is to change it and observe a shift in that function.We will use psilocybin as a pharmacological probe of the serotonin system in vivo. We will directly test the hypothesis that serotonergic targets of psilocybin – principally, but not exclusively, 5HT2A receptor pathways—function differently in autistic and non-autistic adults.MethodsThe ‘PSILAUT’ “shiftability” study is a case–control study autistic and non-autistic adults. How neural responses ‘shift’ in response to low doses (2 mg and 5 mg) of psilocybin compared to placebo will be examined using multimodal techniques including functional MRI and EEG. Each participant will attend on up to three separate visits with drug or placebo administration in a double-blind and randomized order.ResultsThis study will provide the first direct evidence that the serotonin targets of psilocybin function differently in the autistic and non-autistic brain. We will also examine individual differences in serotonin system function.ConclusionsThis work will inform our understanding of the neurobiology of autism as well as decisions about future clinical trials of psilocybin and/or related compounds including stratification approaches.Trial registrationNCT05651126.

Wheat germ agglutinin modified mixed micelles overcome the dual barrier of mucus/enterocytes for effective oral absorption of shikonin and gefitinib.

The combination of shikonin (SKN) and gefitinib (GFB) can reverse the drug resistance of lung cancer cells by affecting energy metabolism. However, the poor solubility of SKN and GFB limits their clinical application because of low bioavailability. Wheat germ agglutinin (WGA) can selectively bind to sialic acid and N-acetylglucosamine on the surfaces of microfold cells and enterocytes, and is a targeted biocompatible material. Therefore, we created a co-delivery micelle system called SKN/GFB@WGA-micelles with the intestinal targeting functions to enhance the oral absorption of SKN and GFB by promoting mucus penetration for nanoparticles via oral administration. In this study, Caco-2/HT29-MTX-E12 co-cultured cells were used to simulate a mucus/enterocyte dual-barrier environment, and HCC827/GR cells were used as a model of drug-resistant lung cancer. We aimed to evaluate the oral bioavailability and anti-tumor effect of SKN and GFB using the SKN/GFB@WGA-micelles system. In vitro and in vivo experimental results showed that WGA promoted the mucus penetration ability of micelles, significantly enhanced the uptake efficiency of enterocytes, improved the oral bioavailability of SKN and GFB, and exhibited good anti-tumor effects by reversing drug resistance. The SKN/GFB@WGA-micelles were stable in the gastrointestinal tract and provided a novel safe and effective drug delivery strategy.

Boundary and distributed optimal control for a population dynamics PDE model with discontinuous in time Galerkin FEM schemes

We consider fully discrete finite element approximations for a semilinear optimal control system of partial differential equations in two cases: for distributed and Robin boundary control. The ecological predator-prey optimal control model is approximated by conforming finite element methods mimicking the spatial part, while a discontinuous Galerkin method is used for the time discretization. We investigate the sensitivity of the solution distance from the target function, in cases with smooth and rough initial data. We employ low, and higher-order polynomials in time and space whenever proper regularity is present. The approximation schemes considered are with and without control constraints, driving efficiently the system to desired states realized using non-linear gradient methods.

The regulation of prolactin secretion and its targeting function of teleost

Prolactin is involved in regulating various physiological activities of vertebrates and is one of the most momentous pituitary hormones. However, not enough attention is currently paid to prolactin, especially in teleost. This paper aims to gather, organize, and analyze recent studies on the regulation and functions of prolactin. By comparing with other animal groups, it highlights the significant role of prolactin in fish reproduction, immunity, growth, and osmotic pressure regulation, as well as the upstream and downstream factors that may be involved in the regulation of prolactin functions were introduced to provide a theoretical basis for the in-depth study and potential practical application of prolactin.

Drug-Online: an online platform for drug-target interaction, affinity, and binding sites identification using deep learning

BackgroundAccurately identifying drug-target interaction (DTI), affinity (DTA), and binding sites (DTS) is crucial for drug screening, repositioning, and design, as well as for understanding the functions of target. Although there are a few online platforms based on deep learning for drug-target interaction, affinity, and binding sites identification, there is currently no integrated online platforms for all three aspects.ResultsOur solution, the novel integrated online platform Drug-Online, has been developed to facilitate drug screening, target identification, and understanding the functions of target in a progressive manner of “interaction-affinity-binding sites”. Drug-Online platform consists of three parts: the first part uses the drug-target interaction identification method MGraphDTA, based on graph neural networks (GNN) and convolutional neural networks (CNN), to identify whether there is a drug-target interaction. If an interaction is identified, the second part employs the drug-target affinity identification method MMDTA, also based on GNN and CNN, to calculate the strength of drug-target interaction, i.e., affinity. Finally, the third part identifies drug-target binding sites, i.e., pockets. The method pt-lm-gnn used in this part is also based on GNN.ConclusionsDrug-Online is a reliable online platform that integrates drug-target interaction, affinity, and binding sites identification. It is freely available via the Internet at http://39.106.7.26:8000/Drug-Online/.

Solving a class of multi-scale elliptic PDEs by Fourier-based mixed physics informed neural networks

Deep neural networks have garnered widespread attention due to their simplicity and flexibility in the fields of engineering and scientific calculation. In this study, we probe into solving a class of elliptic partial differential equations (PDEs) with multiple scales by utilizing Fourier-based mixed physics informed neural networks (dubbed FMPINN), its solver is configured as a multi-scale deep neural network. In contrast to the classical PINN method, a dual (flux) variable about the rough coefficient of PDEs is introduced to avoid the ill-condition of neural tangent kernel matrix caused by the oscillating coefficient of multi-scale PDEs. Therefore, apart from the physical conservation laws, the discrepancy between the auxiliary variables and the gradients of multi-scale coefficients is incorporated into the cost function, obtaining a satisfactory solution of PDEs by minimizing the defined loss through some optimization methods. Additionally, a trigonometric activation function is introduced for FMPINN, which is suited for representing the derivatives of complex target functions. Handling the input data by Fourier feature mapping will effectively improve the capacity of deep neural networks to solve high-frequency problems. Finally, to validate the efficiency and robustness of the proposed FMPINN algorithm, we present several numerical examples of multi-scale problems in various dimensional Euclidean spaces. These examples cover low-frequency and high-frequency oscillation cases, demonstrating the effectiveness of our approach. All code and data accompanying this manuscript will be publicly available at https://github.com/Blue-Giant/FMPINN.

Harnessing the power of new genetic tools to illuminate Giardia biology and pathogenesis.

Giardia is a prevalent single-celled microaerophilic intestinal parasite causing diarrheal disease and significantly impacting global health. Double diploid (essentially tetraploid) Giardia trophozoites have presented a formidable challenge to the development of molecular genetic tools to interrogate gene function. High sequence divergence and the high percentage of hypothetical proteins lacking homology to proteins in other eukaryotes have limited our understanding of Giardia protein function, slowing drug target validation and development. For more than 25 years, Giardia A and B assemblages have been readily amenable to transfection with plasmids or linear DNA templates. Here, we highlight the utility and power of genetic approaches developed to assess protein function in Giardia, with particular emphasis on the more recent clustered regularly interspaced palindromic repeats/Cas9-based methods for knockdowns and knockouts. Robust and reliable molecular genetic approaches are fundamental toward the interrogation of Giardia protein function and evaluation of druggable targets. New genetic approaches tailored for the double diploid Giardia are imperative for understanding Giardia's unique biology and pathogenesis.

Piloting Rasch model scoring of the National Eye Institute Visual Function Questionnaire in uveitis

IntroductionThe National Eye Institute Visual Function Questionnaire (NEI VFQ) is a common patient-reported outcome measure (PROM) in uveitis trials. Its psychometric properties using state-of-the-art scoring based on Rasch models, a latent trait model that improves accuracy of PROMs assessment, has not yet been investigated.MethodsThe study participants were recruited online from uveitis patient organizations, where individuals self-reported their uveitis diagnosis and visual acuity level. These participants then completed the NEI VFQ-25. The visual function (VF) and socioemotional (SE) subscales were psychometrically analysed in terms of item fit, targeting, internal consistency, dimensionality, and differential item functioning (DIF), using Rasch models. Criterion validity was examined based on associations between NEI VFQ person measures and recent visual acuity (VA) levels.ResultsNinety-nine participants recruited online from uveitis patient organizations (68 women, 31 men; mean age 50 ± 15 years; 46.5% self-reported receiving systematic therapy for uveitis, 0.6% NEI VFQ-25 missing data) were included. The mean difficulty of items was lower than the average person ability. None of the items demonstrated misfit to an extent that would induce noise into the measurement. The consistency metrics person reliability and person separation index of the subscales were 0.85 and 2.34 (NEI VFQ-VF), 0.86 and 2.52 (NEI VFQ-SE), respectively. There was no evidence of multidimensionality and none of the items showed DIF by gender. The differences between item and person measures were 1.44 (NEI VFQ-VF) and 1.03 (NEI VFQ-SE). NEI VFQ-25 person measures were significantly lower in participants with visual impairment (all p values ≤ 0.007).ConclusionRasch model-based scoring of the re-engineered NEI VFQ-25 demonstrates acceptable internal consistency, item fit and construct validity for assessing two key domains of quality of life in individuals self-reporting uveitis. The PROM was targeted at a higher level of difficulty than present in our heterogeneous sample.

Synthesis, Screening, and Evaluation of Theranostic Molecular CPCR4-Based Probe Targeting CXCR4.

Chemokines and chemokine receptors are indispensable to play a key role in the development of malignant tumors. As one of the most widely expressed chemokine receptors, chemokine (C-X-C motif) receptor 4 (CXCR4) has been a popular research focus. In most tumors, CXCR4 expression is significantly upregulated. Moreover, integrated nuclide diagnosis and therapy targeting CXCR4 show great potential. [68Ga]Ga-pentixafor, a radioligand targeting CXCR4, exhibits a strong affinity for CXCR4 both in vivo and in vitro. However, [177Lu]Lu-pentixather, the therapeutic companion of [68Ga]Ga-pentixafor, requires significant refinement to mitigate its pronounced hepatic biodistribution. The objective of this study was to synthesize theranostic molecular tracers with superior CXCR4 targeting functions. The Daudi cell line, which highly expressed CXCR4, and the MM.1S cell line, which weakly expressed CXCR4, were used in this study. Based on the pharmacophore cyclo (-d-Tyr-n-me-d-Orn-l-Arg-L-2-NAL-Gly-) (CPCR4) of pentixafor, six tracers were synthesized: [124I]I-1 ([124I]I-CPCR4), [99mTc]Tc-2 ([99mTc]Tc-HYNIC-CPCR4), [124I]I-3 ([124I]I-pentixafor), [18F]AlF-4 ([18F]AlF-NETA-CPCR4), [99mTc]Tc-5 ([99mTc]Tc-MAG3-CPCR4) and [124I]I-6 ([124I]I-pentixafor-Ga) and their radiochemical purities were all higher than 95%. After positron emission tomography (PET)/single-photon emission computed tomography (SPECT) imaging, the [124I]I-6 group exhibited the best target-nontarget ratio. At the same time, comparing the [68Ga]Ga-pentixafor group with the [124I]I-6 group, we found that the [124I]I-6 group had a better target-nontarget ratio and lower uptake in nontarget organs. Therefore, compound 6 was selected for therapeutic radionuclide (131I) labeling, and the tumor-bearing animal models were treated with [131I]I-6. The volume of the tumor site was significantly reduced in the treatment group compared with the control group, and no significant side effects were found. [124I]I-6 and [131I]I-6 showed excellent affinity for targeting CXCR4, and they showed great potential for the integrated diagnosis and treatment of tumors with high CXCR4 expression.

Integrated design optimization method for pavement structure and materials based on further development of finite element and particle swarm optimization algorithm

The maintenance design for existing asphalt pavements, characterized by their typical structure and material composition, falls short of meeting the diverse requirements of pavements. There is an urgent need for a more rational design approach that can achieve accurate pavement design while also reducing construction costs. This paper focuses on selected conventional road surface structures and conducts an analysis of how pavement structure and materials impact fatigue life and construction expenses. It employs Python to perform three-dimensional finite element road modeling, analysis, and data extraction at specific locations, using normalized fatigue equation to calculate pavement fatigue life. The optimization variables selected are the modulus E and thickness h, with pavement maintenance cost serving as the optimization target function. The particle swarm optimization algorithm is employed for optimizing these variables. A comparative analysis is carried out for five improved optimization algorithms, optimizing combinations of inertia weight, learning factors, particle speed, number of particles, and iteration times. The results reveal that the enhanced optimization algorithms significantly improve iteration trends, featuring velocity limitations and natural selection mechanisms. Between 0 and 20 iterations, all algorithms search for the optimal solution over a wider interval with greater fluctuations. Between 20 and 30 iterations, each algorithm gradually converges towards its solution, stabilizing in the later stages. Based on optimization speed and actual engineering cost benchmarks, the NDPSO algorithm is selected, resulting in a 52.8% reduction in actual engineering costs. Adjustments to the particle count and iteration number of the NDPSO algorithm are made based on computational cost and optimization accuracy, thus choosing 20 particles and 50 iterations as the recommended parameter combination. By combining automated finite element modeling, analysis, and particle swarm optimization, this paper offers a novel and effective automated design reference for extending the lifespan of asphalt pavement structures.

Redirect Tropism of Fowl Adenovirus 4 Vector by Modifying Fiber2 with Variable Domain of Heavy-Chain Antibody.

The variable domain of a heavy-chain antibody (VHH) has the potential to be used to redirect the cell tropism of adenoviral vectors. Here, we attempted to establish platforms to simplify the screening of VHHs for their specific targeting function when being incorporated into the fiber of adenovirus. Both fowl adenovirus 4 (FAdV-4) and simian adenovirus 1 (SAdV-1) have two types of fiber, one of which is dispensable for virus propagation and is a proper site for VHH display. An intermediate plasmid, pMD-FAV4Fs, was constructed as the start plasmid for FAdV-4 fiber2 modification. Foldon from phage T4 fibritin, a trigger for trimerization, was employed to bridge the tail/shaft domain of fiber2 and VHHs against human CD16A, a key membrane marker of natural killer (NK) cells. Through one step of restriction-assembly, the modified fiber2 was transferred to the adenoviral plasmid, which was linearized and transfected to packaging cells. Five FAdV-4 viruses carrying the GFP gene were finally rescued and amplified, with three VHHs being displayed. One recombinant virus, FAdV4FC21-EG, could hardly transduce human 293 or Jurkat cells. In contrast, when it was used at a multiplicity of infection of 1000 viral particles per cell, the transduction efficiency reached 51% or 34% for 293 or Jurkat cells expressing exogenous CD16A. Such a strategy of fiber modification was transplanted to the SAdV-1 vector to construct SAdV1FC28H-EG, which moderately transduced primary human NK cells while the parental virus transduced none. Collectively, we reformed the strategy of integrating VHH to fiber and established novel platforms for screening VHHs to construct adenoviral vectors with a specific tropism.

ALYREF m5C RNA methylation reader predicts bladder cancer prognosis by regulating the tumor immune microenvironment.

5-Methylcytidine (m5C) methylation is a recently emerging epigenetic modification that is closely related to tumor proliferation, occurrence, and metastasis. This study aimed to investigate the clinicopathological characteristics and prognostic value of m5C regulators in bladder cancer (BLCA), and their correlation with the tumor immune microenvironment. Thirteen m5C RNA methylation regulators were analyzed using RNA-sequencing and corresponding clinical information obtained from the TCGA database. The Cluster Profiler package was used to analyze the gene ontology function of potential targets and enriched the Kyoto Encyclopedia of Genes and Genomes pathway. Kaplan-Meier survival analysis was used to compare survival differences using the log-rank test and univariate Cox proportional hazards regression. The correlation between signature prognostic m5C regulators and various immune cells was analyzed. Univariate and multivariate Cox regression analyses identified independence of the ALYREF gene signature. Nine out of the 13 m5C RNA methylation regulators were differentially expressed in BLCA and normal samples and were co-expressed. These 9 regulators were associated with clinicopathological tumor characteristics, particularly high or low tumor risk, pT or pTNM stage, and migration. Consensus clustering analysis divides the BLCA samples into 4 clusters. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment annotation and gene ontology function analysis identified 273 upregulated and 594 downregulated genes in BLCA. Notably, only ALYREF was significantly correlated with OS (P < .05). ALYREF exhibited significant infiltration levels in macrophage cells. Therefore, we constructed a nomogram for ALYREF as an independent prognostic factor. Additionally, we observed that both the mRNA and protein levels of ALYREF were upregulated, and immunofluorescence showed that ALYREF was mainly distributed in nuclear speckles. ALYREF overexpression was significantly associated with poor OS. Our findings demonstrated the potential of ALYREF to predict clinical prognostic risks in BLCA patients and regulate the tumor immune microenvironment. As such, ALYREF may serve as a novel prognostic indicator in BLCA patients.