Complete Characterization Research Articles

Frontal Maintenance in Submesoscale Flows

Abstract Classic deformation theory includes parameters – divergence, total strain, and vorticity – that are invariant to changes in the coordinate system. However, these parameters are sometimes ambiguous with respect to characterizing how fronts are formed and maintained, because the presence of a front imposes a reference coordinate system. To help remedy this ambiguity, we propose a framework in frontal coordinates based on along- and cross-front velocity gradients to better characterize frontal maintenance, which can also be used to define divergence and normal strain in frontal coordinates. The framework with these four parameters defines eight characteristic flowtypes at a front, providing a complete characterization of the flowthat strengthens or weakens a front. This framework highlights the importance of the ‘strain efficiency’ concept, which unambiguously defines the contribution of total strain to frontogenesis. Two examples, one based on a realistic simulation of submesoscales in the northern Gulf of Mexico and the other based on an idealized model with similar flow characteristics, are provided to demonstrate how this framework can be used to enhance our understanding frontal dynamics in submesoscale flows.

Recognizing integrality of weighted rectangles partitions

Given a grid of active and inactive pixels, the weighted rectangles partitioning (WRP) problem is to find a maximum-weight partition of the active pixels into rectangles. WRP is formulated as an integer programming problem and instances with an integral relaxation polyhedron are characterized by a balanced problem matrix. A complete characterization of these balanced instances is proved. In addition, computational results on balancedness recognition and on solving WRP are presented.

Atypical implicit and explicit sense of agency in autism: A complete characterization using the cue integration approach.

There exist indications that sense of agency (SoA), the experience of being the cause of one's own actions and actions' outcomes, is altered in autism. However, no studies in autism have simultaneously investigated the integration mechanisms underpinning both implicit and explicit SoA, the two levels of agency proposed by the innovative cue integration approach. Our study establishes a first complete characterization of SoA functioning in autism, by comparing age- and IQ-matched samples of autistic versus neurotypical adults. Intentional binding and judgments of agency were used to assess implicit and explicit SoA over pinching movements with visual outcomes. Sensorimotor and contextual cues were manipulated using feedback alteration and induced belief about the cause of actions' outcome. Implicit SoA was altered in autism, as showed by an overall abolished intentional binding effect and greater inter-individual heterogeneity. At the explicit level, we observed under-reliance on retrospective sensorimotor cues. The implicit-explicit dynamic was also altered in comparison to neurotypical individuals. Our results show that both implicit and explicit levels of SoA, as well as the dynamic between the two levels, present atypicalities in autism.

Underexplored rare bacterial biosphere associated with corals of the northeastern Tropical Eastern Pacific

Microbes of low abundance (i.e., rare biosphere) are involved in the ability of the coral as holobiont, to respond and acclimatize to local and global environmental disturbances. However, no complete characterization of these important microbial assemblages has been conducted. Outlining these assemblages would greatly benefit our understanding of their ecological role in the coral-microbial association. Here, we assessed the rare bacterial biosphere of the hermatypic corals Pocillopora cf. damicornis and P. cf. verrucosa, as well as the surrounding seawater and sediment, at six sites in the northeastern Tropical Eastern Pacific, using MiSeq Illumina sequencing of the 16S rRNA gene region V4. The rare biosphere presented spatial differences among sites with different anthropogenic stress levels. The four biotopes presented a similar rare microbiota. However, in sites with high anthropogenic stress, a high abundance of the Vibrionaceae family was recorded, while locations with low and moderate anthropogenic stress showed several high sequences related to the Flavobacteriaceae and Bacillaceae families. This study represents the first approach to the knowledge of the rare bacterial biosphere in P. cf. damicornis and P. cf. verrucosa in the northeast of the Eastern Tropical Pacific and its possible use as a bioindicator of the stress response to the magnitude of anthropogenic stress.

Complete Sequence and Characterization of Mitochondrial and Chloroplast Genome of Navicula incerta CACC 0356.

Navicula incerta, a marine benthic diatom, holds promise for human nutrition and health as well as for aquaculture applications. However, the scarcity of organelle genome data within the Navicula clade has impeded a comprehensive understanding and utilization of this group. Our research presents a pioneering exploration into the complete mitochondrial and chloroplast genome sequences of N. incerta CACC 0356, shedding light on its phylogeny and evolutionary history. The mitochondrial genome (mtDNA) spans 61,320 bp with a GC content of 29.87%, comprising one circular DNA molecule that encodes for 34 protein-coding genes, 24 tRNAs, and 34 rRNAs. Whereas, the chloroplast genome (cpDNA) is larger at 129,090 bp, encompassing 127 protein-coding genes, 30 tRNAs, and 7 rRNAs. Notably, the cpDNA of N. incerta is approximately 2.1 times the size of its mtDNA. Our annotation identified four genes that are partially situated in the homologous regions between the mitogenome and chloroplast genome, totaling 372 bp, which represents 0.61% of the entire mitogenome. Comparative analyses revealed that N. incerta CACC 0356 is closely affiliated with Fistulifera saprophila and Fistulifera solaris, both members of the Naviculaceae order. This study significantly expands the mitochondrial and chloroplast genomic resources for algae and lays a foundation for the development of genetic diversity analyses in algae.

Division quaternion algebras over some cyclotomic fields

Let p 1, p 2 be two distinct prime integers, let n be a positive integer, n ≥ 3 and let ξn be a primitive root of order n of the unity. In the 3rd section of this paper we obtain a complete characterization for a quaternion algebra H ( p 1 , p 2 ) to be a division algebra over the nth cyclotomic field Q ( ξ n ) , when n ∈ { 3 , 4 , 6 , 7 , 8 , 9 , 11 , 12 } and we also obtain a characterization for a quaternion algebra H ( p 1 , p 2 ) to be a division algebra over the nth cyclotomic field Q ( ξ n ) , when n ∈ { 5 , 10 } . In the 4th section we obtain a complete characterization for a quaternion algebra H Q ( ξ n ) ( p 1 , p 2 ) to be a division algebra, when n = l k , with l a prime integer, l ≡ 3 (mod 4) and k a positive integer. In the last section of this article we obtain a complete characterization for a quaternion algebra H Q ( ξ l ) ( p 1 , p 2 ) to be a division algebra, when l is a Fermat prime number.

Effect of Dynamic Corrosion and Degradation on Fatigue Life of an AA2024-T3 Aircraft Profile: Experimental, Statistical, and Numerical Analyses

The effect of parameters such as angle of attack, flow velocity, and electrolyte concentration (saline solution) on the extent of material degradation, as well as the morphology and depth of corrosion pits in an airfoil made of 2024-T3 aluminum alloy, was studied in detail. An orthogonal L9 design of experiments (Taguchi method) was applied to promote high pitting corrosion through the quality characteristic “the higher the better”. Potentiodynamic curves of the three experiments with low, medium, and high saline concentration were obtained through the CorrTest CS350 equipment. The above allowed the determination of the electrochemical corrosion parameters under static test conditions. The corroded airfoils were analyzed using light optical microscopy (LOM). In addition, the roughness measurements correlated with the extent of the degraded surface. A complete pit shape and depth characterization was obtained by applying mechanical ground, surface wear level monitoring (every 0.01 mm), and LOM observations. Pitting defects (depth and morphology) and mechanical strength reduction were considered by a finite element (FE) model to simulate the airfoil fatigue behavior. Numerical results helped to determine the contribution of pitting and the extent of degradation on sudden airfoil failure.

CASE (CemiplimAb-rwlc Survivorship and Epidemiology): a study in advanced basal cell carcinoma.

Patients diagnosed with metastatic basal cell carcinoma (BCC) have a poor prognosis. The current standard of care for adults with locally advanced or metastatic BCC who are not candidates for surgery or radiation therapy is treatment with hedgehog pathway inhibitors (HHIs). For patients who progress while on this therapy, further treatment options are limited. There is also a need for real-world clinical practice data on the clinical characteristics, management, disease progression, and survivorship of these patients. The ongoing CemiplimAb-rwlc Survivorship and Epidemiology (CASE) study is a phase IV, multicenter, prospective, noninterventional survivorship and epidemiology cohort study evaluating the effectiveness and safety of cemiplimab, a fully human immunoglobulin G4 monoclonal antibody that blocks the interaction between the programmed cell death-1 (PD-1) receptor and its ligands. This paper describes one cohort of the CASE study of patients with locally advanced or metastatic BCC who have failed or are intolerant of HHIs or for whom HHI therapy is not appropriate. Outcome measures of the study include response to treatment, quality of life, safety, treatment patterns, patient experience, and survival. This study could provide a more complete characterization of this patient population and fill knowledge gaps related to real-world treatment utilization and patient outcomes.Clinical Trial registration: NCT03836105.

I-motif formation in the promoter region of the B-MYB proto-oncogene.

Understanding the mechanisms of carcinogenesis is essential to combat cancer. The search for alternative targets for anticancer therapy has gained interest, particularly when focused on upstream pathways. This strategy is particularly relevant when the encoded target proteins are known - or believed - to be "undruggable", as has been reported for the B-MYB oncogene. This gene, which regulates survival and cell cycle regulation, is overexpressed in cancer and correlates with an unfavorable prognosis. In this study, we focused on the identification of the i-motif (iM) structures in the promoter region of B-MYB as a possible anticancer target, with a complete biophysical characterization and in cell formation assessment using iM-CUT&Tag. Additionally, the interaction of the iM structures with a library of small molecules was investigated.

Galaxy @Sciensano: a comprehensive bioinformatics portal for genomics-based microbial typing, characterization, and outbreak detection

The influx of whole genome sequencing (WGS) data in the public health and clinical diagnostic sectors has created a need for data analysis methods and bioinformatics expertise, which can be a bottleneck for many laboratories. At Sciensano, the Belgian national public health institute, an intuitive and user-friendly bioinformatics tool portal was implemented using Galaxy, an open-source platform for data analysis and workflow creation. The Galaxy @Sciensano instance is available to both internal and external scientists and offers a wide range of tools provided by the community, complemented by over 50 custom tools and pipelines developed in-house. The tool selection is currently focused primarily on the analysis of WGS data generated using Illumina sequencing for microbial pathogen typing, characterization and outbreak detection, but it also addresses specific use cases for other data types. Our Galaxy instance includes several custom-developed 'push-button' pipelines, which are user-friendly and intuitive stand-alone tools that perform complete characterization of bacterial isolates based on WGS data and generate interactive HTML output reports with key findings. These pipelines include quality control, de novo assembly, sequence typing, antimicrobial resistance prediction and several relevant species-specific assays. They are tailored for pathogens with active genomic surveillance programs, and clinical relevance, such as Escherichia coli, Listeria monocytogenes, Salmonella spp. and Mycobacterium tuberculosis. These tools and pipelines utilize internationally recognized databases such as PubMLST, EnteroBase, and the NCBI National Database of Antibiotic Resistant Organisms, which are automatically synchronized on a regular basis to ensure up-to-date results. Many of these pipelines are part of the routine activities of Belgian national reference centers and laboratories, some of which use them under ISO accreditation. This resource is publicly available for noncommercial use at https://galaxy.sciensano.be/ and can help other laboratories establish reliable, traceable and reproducible bioinformatics analyses for pathogens encountered in public health settings.

Developing a Novel Agrochemical-Based MOF: A Multifunctional Platform with Herbicidal and Antibacterial Activities.

Excessive and uncontrolled application of agrochemicals has resulted in contamination of terrestrial and aquatic environments. In the past decade, metal-organic frameworks (MOFs) have been studied as agrochemical release systems to enhance efficiency while reducing the leaching of toxic molecules to the environment. In this work, we take a further step and use organic agrochemicals as linkers in the preparation of MOFs, which we have called AgroMOFs. Particularly, we have prepared a novel 2D framework, named GR-MOF-20, based on the flexible herbicide glyphosine (H5Gly) and the widely used antibacterial and fungicide Cu2+. After complete characterization by single-crystal and powder X-ray diffraction and high-performance liquid chromatography (HPLC), a fascinating transformation of the structure to a novel 3D-MOF (GR-MOF-21) after in water is observed, together with a burst degradation (86.0 ± 3.2% after 72 h). Considering the bioactivity of its building blocks (H5Gly and Cu2+), the combined herbicidal and antibacterial activities of GR-MOF-21 were demonstrated. Aqueous solutions prepared with GR-MOF-20 (450 mg L-1) demonstrated a better inhibition effect (21.7 ± 2.9% after 7 days) in the germination of seeds of the invasive weed Lolium multiflorum than the free H5Gly (13.3 ± 2.9%). Further, the selectivity of GR-MOF-21 was tested against a nontarget plant (wheat, Triticum aestivum). Finally, GR-MOF-21 showed an important antibacterial activity against Escherichia coli and Pseudomonas syringae, achieving the results obtained with Cu2+ solutions. These observations demonstrated that the use of herbicide and antibacterial units in the MOF construction is an efficient approach to controlling bacteria and weed plants without damaging wheat crops.

Nanotubular clay minerals for simultaneous sorption of pesticides and PFCAs: a molecular simulation study.

Perfluoroalkyl and polyfluoroalkyl substances (PFAS) and herbicides are important persistent contaminants that require specific management. A variety of herbicides is stored in fluorinated containers in the form of aquatic solutions. In such environments, the simultaneous release of PFAS and herbicides takes place. Nature-based solutions, such as the use of clay materials as possible sorbents, are attractive for the immobilization of such contaminants and environmental protection. Nanotubular clay minerals, such as halloysite and imogolite, are sufficient sorbents for herbicides. Due to their structural morphology, such materials could be efficient sorbents for the simultaneous immobilization of PFAS and herbicides. In this study, the potential sorption of a short chain PFAS, perfluorobutanoic acid (PFBA), sorbent of PFBA, and herbicides (atrazine and diuron) were investigated. Forcefield calculations were used for the classical molecular simulation study. Different distributions, arrangements, and ratios of the investigated molecules were investigated for the complete structural and energy characterization of the systems. Both clay minerals created stable complexes with PFBA as well as with both PFBA and herbicide molecules. Halloysite mineral led to similar total energies of the system with sorbed PFBA molecules alone, herbicides alone, or both of the pollutants. In contrast, imogolite led to lower energies with sorbed herbicides and showed relatively higher energies when interacting with PFBA. The complexes with both of the pollutants presented moderate energies. Electrostatic interactions were dominant in all the investigated complexes.

Comprehensive, Quantitative Analysis of SRM 1950: the NIST Human Plasma Reference Material.

Many analytical methods have been developed for performing targeted metabolomics. By combining multiple analytical techniques, comprehensive coverage of the metabolome can be achieved. We combined multiple analytical techniques to comprehensively and quantitatively characterize the widely studied NIST human plasma reference material, SRM 1950. Our goal was to provide a large, well-validated list of confident metabolite concentration values (i.e., benchmarks) to assist the metabolomics community in its calibration and comparison efforts. We used four analytical platforms: high-resolution NMR spectroscopy, direct injection tandem MS (DI-MS/MS), liquid chromatography tandem MS (LC-MS/MS), and inductively coupled plasma MS (ICP-MS). Eight validated analytical assays were run, yielding accurate quantitative measurements for 728 unique metabolites or metabolite species. Through computer-aided literature mining, we identified another 330 unique metabolites previously quantified in SRM 1950. We compared NIST-certified values along with literature-derived concentrations/ranges to the metabolite concentrations measured by our four platforms and eight assays. From these assays/platforms, we generated a list of high-confidence concentration values of 1058 metabolites or metabolite species in SRM 1950 including data for 60 amino acids/related compounds, 48 bile acids, 72 amines/sugars/alcohols, 21 metals, 8 catecholamines, 11 vitamins, 92 organic acids, 40 fatty acids/steroids/nucleobases/indole derivatives, 5 polyfluorinated compounds, 7 carotenoids, 39 acylcarnitines, 76 oxylipins, 13 sterols, and 566 lipids/lipid species. This data set represents the most complete quantitative characterization of SRM 1950. An online database (SRM1950-DB) containing 1058 plasma metabolites/metabolite species in SRM 1950, their structures, HMDB IDs, mass, chemical class, concentrations, references, and reliability is freely available at https://srm1950-data.wishartlab.com.

RHEOLOGY OF FRESH CONCRETE

Much is known about the rheology of particulate suspensions, including cement paste and concrete. Recent studies of cement paste using specialized rheological techniques have provided considerable information about the relationships between flow behavior and microstructure, including the effects of hydration on yield stress. Concrete workability is routinely characterized using the slump test, but rheometers are available that provide a more complete characterization of flow behavior. This paper presents some preliminary rheology results for concrete, both ordinary concrete and concrete designed as an airport pavement overlay, and discusses how rheological measurements might be used in concrete technology. Many technological advantages are provided by full characterization of flow behavior, particularly that it provides access to the large body of knowledge concerning the flow behavior of suspensions.

Complete Genome Characterization of Canine Adenovirus From Türkiye With Next-Generation Sequencing.

Determining the complete genome sequence data of adenoviruses has recently become greatly important due to their use by scientists as vectors in cancer studies and other fields, including vaccine development. However, the GenBank database currently has few complete genome sequences of adenoviruses, which are known for their large genomes. To address this gap, we analysed next-generation sequencing data obtained from our previous study to provide the complete genome sequence of the canine adenovirus-2 strain. For the obtained canine adenovirus-2 strain (OQ596341), comparative genomics, recombination and phylogenetic analysis were conducted. This sequence was compared and phylogenetically analysed with the 20 complete genome sequences of canine adenovirus previously reported in GenBank worldwide, as well as partial E3 ORFA sequences obtained from Türkiye. The nucleotide similarity rates of the sequence obtained from this study with other CAdV-2 whole genomes are over 99.04%. The gene alignment results reveal that the OQ596341 was found to be closely related to the AC000020 reference genome and LC557011. There are two recombination events related to the genome in this study. Comparisons with other complete genome sequences revealed several previously unseen mutations. These mutations include H34Y in the E1A gene; P55A in the E1B 55K gene; D13N and D202N in the IVa2 gene; K679R, V934I and K989N in the Pol gene; E205K in the pTP gene; T455A in the pIIIa gene; A310V in the V gene; G151R in the protease gene; E268K in the 100K gene; G66S and G141S in the 33K gene; T14A, E250K, D287N and I293T in the E3 ORFA gene; and L193F in the E434K gene. Moreover, a comparison with partial sequences obtained from Türkiye revealed the E250K mutation in the E3 ORFA gene, which we report for the first time in Türkiye. The complete CAdV-2 genome sequence obtained in the present study is the first sequence from Europe. Comparative analysis with other genomes revealed some unique mutations. This study is the first to report the E250K amino acid change in the E3 ORFA gene in Türkiye. We anticipate that this data can be used in future CAdV-2 vaccine development studies. Further studies are recommended to evaluate the impact of these mutations on viral tropism and other host interactions.

Evolution of the electrical characteristics of the ATLAS18 ITk strip sensors with HL-LHC radiation exposure range

The objective of the study is to evaluate the evolution of the performance of the new ATLAS Inner-Tracker (ITk) strip sensors as a function of radiation exposure, to ensure the proper operation of the upgraded detector during the lifetime of the High-Luminosity Large Hadron Collider (HL-LHC). Full-size ATLAS ITk Barrel Short-Strip (SS) sensors with final layout design, ATLAS18SS, have been irradiated with neutrons and gammas, to confirm the results obtained with prototypes and miniature sensors during the development phase. The irradiations cover a wide range of fluences and doses that ITk will experience, going from 1×1013 neq/cm2 and 0.49 Mrad, to 1.6×1015 neq/cm2 and 80 Mrad. The split irradiation enables a proper combination of fluence and dose values of the HL-LHC, including a 1.5 safety factor. A complete electrical characterization of the key sensor parameters before and after irradiation is presented, studying the leakage current, bulk capacitance, single-strip and inter-strip characteristics. The results confirm the fulfilment of the ATLAS specifications throughout the whole experiment. The study of a wide range of fluences and doses also allows to obtain detailed results, such as the frequency dependence of the bulk capacitance measurements for highly irradiated sensors, or the evolution of the punch-through protection and inter-strip resistance with radiation.

Characterization of the functional component in human milk and identification of the molecular mechanisms undergoing prematurity.

Human milk (HM) is the earliest form of extrauterine communication between mother and infant, that could promote early programming. The aim of this study is to look for specific biological processes, particularly those undergoing prematurity, modulated by proteins and miRNAs of HM that could be implicated in growth and development. This is a prospective, observational, single center study in which we collected 48 human milk (HM) samples at two distinct stages of lactation: colostrum (first 72-96h) and mature milk (at week 4 post-delivery) from mothers of very preterm newborns (<32 weeks) and term (≥37 and<42 weeks). Qualitative and quantitative proteomic and transcriptomic analysis was done in our samples. We performed isolation and characterization of HM extracellular vesicles (EVs) to carry out proteomic and transcriptomic analysis in colostrum (CM) and mature milk (MM). Proteomic analysis revealed a functional role of CM in immunological protection and MM in metabolic processes. TENA, TSP1 and OLF4, proteins with roles in immune response and inflammatory modulation, were upregulated in CM vs MM, particularly in preterm. HM modulation differed depending on gestational age (GA). The miRNAs identified in HM are implicated in structural functions, including growth and neurological development. miRNA-451a was differentially expressed between groups, and downregulated in preterm CM. Because the particularities of each GA are reflected in the EVs content of HM, providing newborns with HM from their own mother is the optimal way for satisfying their specific needs. Although the role of the proteomic profile of CM and MM of different GA in relation to neurodevelopment has been previously described, this is the first study to show a complete functional characterization of HM (proteome, miRNA at the same time), unmasking the molecular mechanisms related to EVs signaling and their functional role in preterm.

Improved production of palitantin by design of experiments and semi-synthesis of palitantin derivatives for bioactivity assessment

A new method for the production and isolation of (+)-palitantin (1) is herein reported, from cultures of the fungal strain Penicillium sp. AMF1a. (+)-Palitantin was isolated in 160 mg/L yield, as an alternative procedure to obtain 1 at a larger scale. The complete spectroscopic characterization, including conformational analysis, is presented for (+)-palitantin (1) and for its derivatives 3-α-palitantol (2), 3-β-palitantol (3), (Z)-palinitrorin (4), (Z)-paliphenin (5), (Z)-palifluorin (6) and (E)-palifluorin (7). The absolute configuration of the palitantin portion was confirmed by X-ray analysis of compound 4. The (4-(trifluoromethyl)benzyl)-hydrazone derivatives (6 and 7) displayed moderate biological activities. (Z)-Palifluorin (6) exhibited moderate antiplasmodial activity, while (E)-palifluorin (7) displayed weak antibacterial activity against E. faecalis and S. aureus, while palitantin itself was inactive in the same bioassays, indicating that the semi-synthesis of nitrogenated derivatives of 1 may be of potential interest to generate bioactive palitantin derivatives.

Adipose tissue eQTL meta-analysis highlights the contribution of allelic heterogeneity to gene expression regulation and cardiometabolic traits.

Complete characterization of the genetic effects on gene expression is needed to elucidate tissue biology and the etiology of complex traits. In the present study, we analyzed 2,344 subcutaneous adipose tissue samples and identified 34,774 conditionally distinct expression quantitative trait locus (eQTL) signals at 18,476 genes. Over half of eQTL genes exhibited at least two eQTL signals. Compared with primary eQTL signals, nonprimary eQTL signals had lower effect sizes, lower minor allele frequencies and less promoter enrichment; they corresponded to genes with higher heritability and higher tolerance for loss of function. Colocalization of eQTLs with genome-wide association study (GWAS) signals for 28 cardiometabolic traits identified 1,835 genes. Inclusion of nonprimary eQTL signals increased discovery of colocalized GWAS-eQTL signals by 46%. Furthermore, 21 genes with ≥2 colocalized GWAS-eQTL signals showed a mediating gene dosage effect on the GWAS trait. Thus, expanded eQTL identification reveals more mechanisms underlying complex traits and improves understanding of the complexity of gene expression regulation.

Experimental approaches to evaluate solid lipid nanoparticle-based drug delivery systems.

Solid lipid nanoparticles (SLNs) are potential drug carriers due to the several advantages they offer. The physicochemical stability of lipid carriers varies significantly due to their diverse compositions and structures. Appropriate analytical methods are required for the complete characterization of SLNs. Physicochemical characterization includes analysis of bulk properties like particle size, size distribution, zeta potential, morphology, stability, polymorphism, crystallinity, and molecular level properties like microenvironments within nanoparticles and their interactions with drugs. Moreover, drug loading, drug entrapment efficiency, and drug release kinetics are essential parameters to evaluate the efficacy of SLNs as drug delivery systems. In addition to testing the physicochemical stability and functionality of SLN formulations, it is essential to investigate their desired actions through in vivo studies, which are beyond the scope of this article. This review briefly discusses the different experimental techniques and their applications in the field of solid lipid nanoparticles. These techniques can also be used to characterize nanostructure lipid carriers, which are second-generation lipid nanoparticles.