Diverse tsunamigenesis triggered by the Hunga Tonga-Hunga Ha'apai eruption.

On the evening of 15 January 2022, the Hunga Tonga-Hunga Ha'apai volcano1 unleashed a violent underwater eruption, blanketing the surrounding land masses in ash and debris2,3. The eruption generated tsunamis observed around the world. An event of this type last occurred in 1883 during the eruption of Krakatau4, and thus we have the first observations of a tsunami from a large emergent volcanic eruption captured with modern instrumentation. Here we show that the explosive eruption generated waves through multiple mechanisms, including: (1) air-sea coupling with the initial and powerful shock wave radiating out from the explosion in the immediate vicinity of the eruption; (2) collapse of the water cavity created by the underwater explosion; and (3) air-sea coupling with the air-pressure pulse that circled the Earth several times, leading to a global tsunami. In the near field, tsunami impacts are strongly controlled by the water-cavity source whereas the far-field tsunami, which was unusually persistent, can be largely described by the air-pressure pulse mechanism. Catastrophic damage in some harbours in the far field was averted by just tens of centimetres, implying that a modest sea level rise combined with a future, similar event would lead to a step-function increase in impacts on infrastructure. Piecing together the complexity of this event has broad implications for coastal hazards in similar geophysical settings, suggesting a currently neglected source of global tsunamis.

PEtOxylated Interferon-α2a Bioconjugates Addressing H1N1 Influenza A Virus Infection.

Influenza A viruses (IAV), including the pandemic 2009 (pdm09) H1N1 or avian influenza H5N1 virus, may advance into more pathogenic, potentially antiviral drug-resistant strains (including loss of susceptibility against oseltamivir). Such IAV strains fuel the risk of future global outbreaks, to which this study responds by re-engineering Interferon-α2a (IFN-α2a) bioconjugates into influenza therapeutics. Type-I interferons such as IFN-α2a play an essential role in influenza infection and may prevent serious disease courses. We site-specifically conjugated a genetically engineered IFN-α2a mutant to poly(2-ethyl-2-oxazoline)s (PEtOx) of different molecular weights by strain-promoted azide-alkyne cyclo-addition. The promising pharmacokinetic profile of the 25 kDa PEtOx bioconjugate in mice echoed an efficacy in IAV-infected ferrets. One intraperitoneal administration of this bioconjugate, but not the marketed IFN-α2a bioconjugate, changed the disease course similar to oseltamivir, given orally twice every study day. PEtOxylated IFN-α2a bioconjugates may expand our therapeutic arsenal against future influenza pandemics, particularly in light of rising first-line antiviral drug resistance to IAV.

Polymer selection impacts the pharmaceutical profile of site-specifically conjugated Interferon-α2a.

Conjugation of poly(ethylene glycol) (PEG) to biologics is a successful strategy to favorably impact the pharmacokinetics and efficacy of the resulting bioconjugate. We compare bioconjugates synthesized by strain-promoted azide-alkyne cycloaddition (SPAAC) using PEG and linear polyglycerol (LPG) of about 20 kDa or 40 kDa, respectively, with an azido functionalized human Interferon-α2a (IFN-α2a) mutant. Site-specific PEGylation and LPGylation resulted in IFN-α2a bioconjugates with improved in vitro potency compared to commercial Pegasys. LPGylated bioconjugates had faster disposition kinetics despite comparable hydrodynamic radii to their PEGylated analogues. Overall exposure of the PEGylated IFN-α2a with a 40 kDa polymer exceeded Pegasys, which, in return, was similar to the 40 kDa LPGylated conjugates. The study points to an expanded polymer design space through which the selected polymer class may result in a different distribution of the studied bioconjugates.

Nanoparticle sizing in the field of nanomedicine: Power of an analytical ultracentrifuge.

Hydrodynamic and light scattering methods are urgently required for accurate characterization of nanoparticles (NPs) in the field of nanomedicine to unveil their sizes and distributions. A fundamental characterization approach in the field of nanomedicines is, next to standard batch dynamic light scattering (DLS) and increasingly more applied (asymmetrical flow) field-flow fractionation (FFF) coupled to multi-angle laser light scattering (MALLS), the utilization of an analytical ultracentrifuge (AUC). Here, we demonstrate the power of an AUC in comparison to batch DLS and FFF-MALLS to decipher, in detail, the size and dispersity of pharma-relevant, commercial and in-house prepared soft matter NPs, suitable for life science applications. In this study, size and dispersity of poly(lactic-co-glycolic acid) (PLGA) NPs and in-house prepared NPs, consisting of the commercially available pharmapolymer Eudragit® E or of a polymer of similar composition synthesized via reversible addition fragmentation chain transfer (RAFT) polymerization, were investigated. Simultaneously, an insight on the presence of the utilized surfactant on the NP formulations, which is usually limited with other techniques, could be achieved by multi-speed experiments with the AUC in one experimental setting. While the repeatability and ruggedness of observations with modern AUC instruments of the newest generation is demonstrated, the results are further underpinned by the classical relations of hydrodynamics. Investigations aiming at hydrodynamic diameters (from DLS) and radii of gyration (from FFF-MALLS) are critically discussed and compared to the repeatable and rugged investigations by an AUC. The latter is proven to provide a self-sufficient experimental approach for NP characterization in the field of nanomedicine based on absolute principles, compares well to FFF-MALLS, and can unravel issues in NP sizing that arise when more common techniques, such as DLS, are used.

Ethoxy acetalated dextran-based nanocarriers accomplish efficient inhibition of leukotriene formation by a novel FLAP antagonist in human leukocytes and blood.

Leukotrienes are pro-inflammatory lipid mediators generated by 5-lipoxygenase aided by the 5-lipoxygenase-activating protein (FLAP). BRP-201, a novel benzimidazole-based FLAP antagonist, inhibits leukotriene biosynthesis in isolated leukocytes. However, like other FLAP antagonists, BRP-201 fails to effectively suppress leukotriene formation in blood, which limits its therapeutic value. Here, we describe the encapsulation of BRP-201 into poly(lactide-co-glycolide) (PLGA) and ethoxy acetalated dextran (Ace-DEX) nanoparticles (NPs), aiming to overcome these detrimental pharmacokinetic limitations and to enhance the bioactivity of BRP-201. NPs loaded with BRP-201 were produced via nanoprecipitation and the physicochemical properties of the NPs were analyzed in-depth using dynamic light scattering (size, dispersity, degradation), electrophoretic light scattering (effective charge), NP tracking analysis (size, dispersity), scanning electron microscopy (size and morphology), UV-VIS spectroscopy (drug loading), an analytical ultracentrifuge (drug release, degradation kinetics), and Raman spectroscopy (chemical attributes). Biological assays were performed to study cytotoxicity, cellular uptake, and efficiency of BRP-201-loaded NPs versus free BRP-201 to suppress leukotriene formation in primary human leukocytes and whole blood. Both PLGA- and Ace-DEX-based NPs were significantly more efficient to inhibit leukotriene formation in neutrophils versus free drug. Whole blood experiments revealed that encapsulation of BRP-201 into Ace-DEX NPs strongly increases its potency, especially upon pro-longed (≥ 5 h) incubations and upon lipopolysaccharide-challenge of blood. Finally, intravenous injection of BRP-201-loaded NPs significantly suppressed leukotriene levels in blood of mice in vivo. These results reveal the feasibility of our pharmacological approach using a novel FLAP antagonist encapsulated into Ace-DEX-based NPs with improved efficiency in blood to suppress leukotriene biosynthesis.

Core-crosslinked, temperature- and pH-responsive micelles: design, physicochemical characterization, and gene delivery application.

Stimuli-responsive block copolymer micelles can provide tailored properties for the efficient delivery of genetic material. In particular, temperature- and pH-responsive materials are of interest, since their physicochemical properties can be easily tailored to meet the requirements for successful gene delivery. Within this study, a stimuli-responsive micelle system for gene delivery was designed based on a diblock copolymer consisting of poly(N,N-diethylacrylamide) (PDEAm) as a temperature-responsive segment combined with poly(aminoethyl acrylamide) (PAEAm) as a pH-responsive, cationic segment. Upon temperature increase, the PDEAm block becomes hydrophobic due to its lower critical solution temperature (LCST), leading to micelle formation. Furthermore, the monomer 2-(pyridin-2-yldisulfanyl)ethyl acrylate (PDSAc) was incorporated into the temperature-responsive PDEAm building block enabling disulfide crosslinking of the formed micelle core to stabilize its structure regardless of temperature and dilution. The cloud points of the PDEAm block and the diblock copolymer were investigated by turbidimetry and fluorescence spectroscopy. The temperature-dependent formation of micelles was analyzed by dynamic light scattering (DLS) and elucidated in detail by an analytical ultracentrifuge (AUC), which provided detailed insights into the solution dynamics between polymers and assembled micelles as a function of temperature. Finally, the micelles were investigated for their applicability as gene delivery vectors by evaluation of cytotoxicity, pDNA binding, and transfection efficiency using HEK293T cells. The investigations showed that core-crosslinking resulted in a 13-fold increase in observed transfection efficiency. Our study presents a comprehensive investigation from polymer synthesis to an in-depth physicochemical characterization and biological application of a crosslinked micelle system including stimuli-responsive behavior.

Solely aqueous formulation of hydrophobic cationic polymers for efficient gene delivery.

Cationic polymers are promising gene delivery vectors due to their ability to bind and protect genetic material. The introduction of hydrophobic moieties into cationic polymers can further improve the vector efficiency, but common formulations of hydrophobic polymers involve harsh conditions such as organic solvents, impairing intactness and loading efficiency of the genetic material. In this study, a mild, aqueous formulation method for the encapsulation of high amounts of genetic material is presented. A well-defined pH-responsive hydrophobic copolymer, i.e. poly((n-butylmethacrylate)-co-(methylmethacrylate)-co-(2-(dimethylamino) ethylmethacrylate)), (PBMD) was synthesized by reversible addition fragmentation chain transfer (RAFT) polymerization. Exploiting the pH-dependent solubility behavior of the polymer, stable pDNA loaded nanoparticles were prepared and characterized using analytical ultracentrifugation (AUC), cryo-transmission electron microscopy (cryo-TEM) and dynamic light scattering (DLS). This novel formulation approach showed high transfection efficiencies in HEK293T cells, while requiring 5- to 10-fold less pDNA compared to linear polyethylenimine (LPEI), in particular at short incubation times and in serum-containing media. Furthermore, the formulation was successfully adopted for siRNA and mRNA encapsulation and the commercially approved polymer Eudragit® E(PO/100). Overall, the aqueous formulation approach, accompanied by a tailor-made hydrophobic polymer and detailed physicochemical and application studies, led to improved gene delivery vectors with high potential for further applications.

Salient features of medical nanoparticles in biological fluids from an analytical ultracentrifuge.

From the perspective of future translation, medical, biodegradable nanoparticles (NPs) have been investigated using an analytical ultracentrifuge in fluids of various complexity, including human serum, in the temperature range of 6 to 40 °C, and timescales relevant for a nanomedical targeting and clearance application. These studies provided salient insights into the integrity and degradation aspects of the NPs, imposed by varying solution environmental conditions. This was enabled by selective monitoring of the targeting dye moiety, cell-specifically directing the NPs to the desired location of interest, i.e. considering a future translative in vivo application. Our study provides experimental insights that are believed to be of key importance to gauge the feasibility of such translative applications in terms of (i) compatibility with patient sera, (ii) timescales of targeting success, and (iii) timescales of desired erosion enabling clearance from the target. All such aspects are provided a priori any in vivo implementation.

In Situ, Quantitative Assessment of Multifunctional Nanoscale Drug Delivery Systems in Human Serum.

The large volume and diversified nanomedicine market, undergoing a rapid growth, relies not only on the creation and applicative exploration of nanocarrier-based medicines showing significant potential, but in particular, demands a quantitative assessment of their physicochemical properties. In this study, we demonstrate the in situ assessment of multifunctional biodegradable nanoparticle (NP) entries as core components of nanoscale drug delivery systems (NDDSs) by making use of analytical ultracentrifugation (AUC). We determine and elucidate the following characteristics of NPs in NDDSs: NP density and size, targeting dye functionality, encapsulated and free drug, surfactant, and also NP drug release dynamics, quantitatively interconnected to NP degradation. In concept, we demonstrate this by multidetection AUC experiments at variable speed and time profiles. We could verify the quantitative and accurate nature of AUC for assessment of NDDSs, that is, also future nanomedicines. This concerns modeled and real life solution application formats such as cell culture media and human serum.

Incentives of Using the Hydrodynamic Invariant and Sedimentation Parameter for the Study of Naturally- and Synthetically-Based Macromolecules in Solution.

The interrelation of experimental rotational and translational hydrodynamic friction data as a basis for the study of macromolecules in solution represents a useful attempt for the verification of hydrodynamic information. Such interrelation originates from the basic development of colloid and macromolecular science and has proven to be a powerful tool for the study of naturally- and synthetically-based, i.e., artificial, macromolecules. In this tutorial review, we introduce this very basic concept with a brief historical background, the governing physical principles, and guidelines for anyone making use of it. This is because very often data to determine such an interrelation are available and it only takes a set of simple equations for it to be established. We exemplify this with data collected over recent years, focused primarily on water-based macromolecular systems and with relevance for pharmaceutical applications. We conclude with future incentives and opportunities for verifying an advanced design and tailored properties of natural/synthetic macromolecular materials in a dispersed or dissolved manner, i.e., in solution. Particular importance for the here outlined concept emanates from the situation that the classical scaling relationships of Kuhn-Mark-Houwink-Sakurada, most frequently applied in macromolecular science, are fulfilled, once the hydrodynamic invariant and/or sedimentation parameter are established. However, the hydrodynamic invariant and sedimentation parameter concept do not require a series of molar masses for their establishment and can help in the verification of a sound estimation of molar mass values of macromolecules.