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The SARS-CoV-2 Nucleocapsid protein (N) performs several functions during the viral lifecycle, including transcription regulation and viral genome encapsulation. We hypothesized that N toggles between these functions via phosphorylation-induced conformational change, thereby altering N interactions with membranes and RNA. We found that phosphorylation changes how biomolecular condensates composed of N and RNA interact with membranes: phosphorylated N (pN) condensates form thin films, while condensates with unmodified N are engulfed. This partly results from changes in material properties, with pN forming less viscous and elastic condensates. The weakening of protein-RNA interaction in condensates upon phosphorylation is driven by a decrease in binding between pN and unstructured RNA. We show that phosphorylation induces a conformational change in the serine/arginine-rich region of N that increases interaction between pN monomers and decreases nonspecific interaction with RNA. These findings connect the conformation, material properties, and membrane-associated states of N, with potential implications for COVID-19 treatment., Competing Interests: Declaration of Interests The authors declare no competing interests.

Dicarboxylate metallosurfactants (AASM), synthesized by mixing N-dodecyl aminomalonate, -aspartate and -glutamate with CaCl 2 , MnCl 2 and CdCl 2 , were characterized by XRD, FTIR, and NMR spectroscopy. Layered structures, formed by metallosurfactants, were evidenced from differential scanning calorimetry and thermogravimetric analyses. Solvent-spread monolayer of AASM in combination with soyphosphatidylcholine (SPC) and cholesterol (CHOL) were studied using Langmuir surface balance. With increasing mole fraction of AASM mean molecular area increased and passed through maxima at ~60 mol% of AASMs, indicating molecular packing reorganization. Systems with 20 and 60 mol% AASM exhibited positive deviations from ideal behavior signifying repulsive interaction between the AASM and SPC, while synergistic interactions were established from the negative deviation at other combinations. Dynamic surface elasticity increased with increasing surface pressure signifying formation of rigid monolayer. Transition of monolayer from gaseous to liquid expanded to liquid condensed state was established by Brewster angle microscopic studies. Stability of the hybrid vesicles, formed by AASM+SPC+CHOL, were established by monitoring their size, zeta potential and polydispersity index values over 100 days. Size and spherical morphology of hybrid vesicles were confirmed by transmission electron microscopic studies. Biocompatibility of the hybrid vesicles were established by cytotoxicity studies revealing their possible applications in drug delivery and imaging., (© 2024 Wiley-VCH GmbH.)

The effects of two ionic liquids (ILs), 1-butyl-3-methylimidazolium tetrafluoroborate ([bmim]BF 4 ) and 1-butyl-1-methyl pyrrolidinium tetrafluoroborate ([bmp]BF 4 ), on a mixture of phospholipids (PLs) 1,2-dipalmitoyl- sn -glycero-3-phosphatidylcholine (DPPC), 1,2-dipalmitoyl- sn -glycero-3-phosphoethanolamine (DPPE), and 1,2-dipalmitoyl- sn -glycero-3-phosphoglycerol (DPPG) (6:3:1, M/M/M, 70% PL) in combination with 30 mol % cholesterol (CHOL) were investigated in the form of a solvent-spread monolayer and bilayer (vesicle). Surface pressure (π)-area ( A ) isotherm studies, using a Langmuir surface balance, revealed the formation of an expanded monolayer, while the cationic moiety of the IL molecules could electrostatically and hydrophobically bind to the PLs on the palisade layer. Turbidity, dynamic light scattering (size, ζ-potential, and polydispersity index), electron microscopy, small-angle X-ray/neutron scattering, fluorescence spectroscopy, and differential scanning calorimetric studies were carried out to evaluate the effects of IL on the structural organization of bilayer in the vesicles. The ILs could induce vesicle aggregation by acting as a "glue" at lower concentrations (<1.5 mM), while at higher concentrations, the ILs disrupt the bilayer structure. Besides, ILs could result in the thinning of the bilayer, evidenced from the scattering studies. Steady-state fluorescence anisotropy and lifetime studies suggest asymmetric insertion of ILs into the lipid bilayer. MTT assay using human blood lymphocytes indicates the safe application of vesicles in the presence of ILs, with a minimal toxicity of up to 2.5 mM IL in the dispersion. These results are proposed to have applications in the field of drug delivery systems with benign environmental impact.

Biomolecular condensates arising from liquid-liquid phase separation contribute to diverse cellular processes, such as gene expression. Partitioning of client molecules into condensates is critical to regulating the composition and function of condensates. Previous studies suggest that client size limits partitioning, with dextrans >5 nm excluded from condensates. Here, we asked whether larger particles, such as macromolecular complexes, can partition into condensates based on particle-condensate interactions. We sought to discover the biophysical principles that govern particle inclusion in or exclusion from condensates using polymer nanoparticles with tailored surface chemistries as models of macromolecular complexes. Particles coated with polyethylene glycol (PEG) did not partition into condensates. We next leveraged the PEGylated particles as an inert platform to which we conjugated specific adhesive moieties. Particles functionalized with biotin partitioned into condensates containing streptavidin, driven by high-affinity biotin-streptavidin binding. Oligonucleotide-decorated particles exhibited varying degrees of partitioning into condensates, depending on condensate composition. Partitioning of oligonucleotide-coated particles was tuned by altering salt concentration, oligonucleotide length, and oligonucleotide surface density. Remarkably, beads with distinct surface chemistries partitioned orthogonally into immiscible condensates. Based on our experiments, we conclude that arbitrarily large particles can controllably partition into biomolecular condensates given sufficiently strong condensate-particle interactions, a conclusion also supported by our coarse-grained molecular dynamics simulations and theory. These findings may provide insights into how various cellular processes are achieved based on partitioning of large clients into biomolecular condensates, as well as offer design principles for the development of drug delivery systems that selectively target disease-related biomolecular condensates., Competing Interests: Competing interests: The authors declare no competing interest.

Understanding the relationship between a polypeptide sequence and its phase separation has important implications for analysing cellular function, treating disease and designing novel biomaterials. Several sequence features have been identified as drivers for protein liquid-liquid phase separation (LLPS), schematized as a 'molecular grammar' for LLPS. Here we further probe how sequence modulates phase separation and the material properties of the resulting condensates, targeting sequence features previously overlooked in the literature. We generate sequence variants of a repeat polypeptide with either no charged residues, high net charge, no glycine residues or devoid of aromatic or arginine residues. All but one of 12 variants exhibited LLPS, albeit to different extents, despite substantial differences in composition. Furthermore, we find that all the condensates formed behaved like viscous fluids, despite large differences in their viscosities. Our results support the model of multiple interactions between diverse residue pairs-not just a handful of residues-working in tandem to drive the phase separation and dynamics of condensates., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)

We report here the hot carrier (HC) cooling time scales within polyhedral CsPbBr 3 nanocrystals (NCs) characterized by different numbers of facets (6 to 26) utilizing a femtosecond upconversion setup. Interestingly, the observed cooling time scale slows many-fold (>10 times) upon opening the new facets on the NC surface. Furthermore, a temperature-dependent study reveals that cooling in multifaceted NCs is polaron mediated, where newly opened polar facets and the soft lattice of CsPbBr 3 NCs play pivotal roles. Our hallmark result of slow cooling in polyhedral NCs renders an excellent opportunity for harvesting high-energy carriers by a carefully chosen molecular system. To this end, employing the hole scavenger molecule aniline, we successfully extracted hot holes from optically pumped NCs. We believe that several intriguing properties of the polyhedral NCs, including rapid polaron formation, defect-tolerant nature, and the capability of soft lattice to support slow diffusion of charge carriers, resulted in decelerated cooling.

Background: Antibiotic resistances of pathogens and breast cancer warrant the search for new alternative strategies. Phytoextracts can eradicate microbe-borne diseases as well as cancer with lower side effects compared to conventional antibiotics., Aim: Unripe and ripe Azadirachta indica (neem) seed extracts were explored as potential antibiofilm and anticancer agents in combating multidrug-resistant infectious bacteria as well as anticancer agents against the MDR breast cancer cell lines., Methods: Shed-dried neem seeds (both unripe and ripe) were pulverized and extracted using methanol. The chemical components were identified with FTIR and gas chromatography - mass spectrometry. Antibiofilm activity of neem seed extracts were assessed in terms of minimum biofilm inhibitory concentration (MBIC), minimum biofilm eradication concentration (MBEC), and fluorescence microscopic studies on Staphylococcus aureus and Vibrio cholerae. Bacterial cells were studied by fluorescence microscopy using acridine orange/ethidium bromide as the staining agents. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values were evaluated to observe the antibacterial activities. Cytotoxicity of the extracts against human blood lymphocytes and the anticancer activity against drug-resistant breast cancer cell lines were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting (FACS) studies., Results: 4-Ethyl-2-hydroxy-2-cyclopentene-1-one, phthalic acid, and 2-hexyl-tetrahydro thiophane were the major compounds in unripe neem seed, whereas 3,5-dihydroxy-6-methyl-2,3-dihydro-4-H-pyran-4-one and 4-ethylbenzamide were predominant in ripe neem seed. Triazine derivatives were also common for both the extracts. MBIC values of unripe and ripe neem seed extracts for S. aureus are 75 and 100 µg/mL, respectively, and for V. cholerae, they are 100 and 300 µg/mL, respectively. MBEC values of unripe and ripe seed extracts are 500 and 300 µg/mL, respectively for S. aureus and for V. cholerae the values are 700 and 500 µg/mL, respectively. Fluorescence microscopic studies at 16 and 24 h, after bacterial culture, demonstrate enhanced antibiofilm activity for the ripe seed extract than that of the unripe seeds for both the bacteria. MTT assay reveals lower cytotoxicity of both the extracts towards normal blood lymphocytes, and anticancer activity against breast cancer cell line (MDA-MB-231) with superior activity of ripe seed extract. FACS studies further supported higher anticancer activity for ripe seed extract., Conclusions: Methanolic extract of neem seeds could substantially inhibit and eradicate biofilm along with their potent antibacterial and anticancer activities. Both the extracts showed higher antibiofilm and antibacterial activity against S. aureus (gram-positive) than V. cholerae (gram-negative). Moreover, ripe seed extract showed higher antibiofilm and anticancer activity than unripe extracts., (© 2022. The Author(s).)

Dicarboxylic amino acid-based surfactants (N-dodecyl derivatives of -aminomalonate, -aspartate, and -glutamate) in combination with hexadecyltrimethylammonium bromide (HTAB) form a variety of aggregates. Composition and concentration-dependent mixtures exhibit liquid crystal, gel, precipitate, and clear isotropic phases. Liquid crystalline patterns, formed by surfactant mixtures, were identified by polarizing optical microscopy. FE-SEM studies reveal the existence of surface morphologies of different mixed aggregates. Phase transition and associated weight loss were found to depend on the composition where thermotropic behaviours were revealed through combined differential scanning calorimetry and thermogravimetric studies. Systems comprising more than 60 mol% HTAB demonstrate shear-thinning behaviour. Gels cause insignificant toxicity to human peripheral lymphocytes and irritation to bare mouse skin; they do not display the symptoms of cutaneous irritation, neutrophilic invasion, and inflammation (erythema, edema, and skin thinning) as evidenced by cumulative irritancy index score. Gels also exhibit substantial antibacterial effects on Staphylococcus aureus, a potent causative agent of skin and soft tissue infections, suggesting its possible application as a vehicle for topical dermatological drug delivery., (© 2021. The Author(s).)

Aggregation studies of anionic surfactant sodium dodecyl sulphate (SDS) was investigated in aqueous 1-butyl-3-methylimidazolium chloride [bmim]Cl and N-butyl-N-methyl pyrrolidinium tetrafluoroborate [bmp]BF 4 ionic liquid (IL) solutions respectively. Systems were studied by surface tension, conductance, UV-VIS absorption/emission spectroscopy and dynamic light scattering. Critical micelle concentration (CMC) values gradually decreased with increasing IL concentration which indicates synergistic interaction between ILs and SDS. Gibbs free energy change results demonstrated spontaneous micellization induced by ILs; however the effect of ILs were not similar to the corresponding regular salts (NaCl and NaBF 4 ). Aggregation number (n) of micelles, determined by fluorescence quenching method, indicate that the 'n' values increase with increasing ILs concentration, induced by the oppositely charged IL cation. Size of the micelles, determined by dynamic light scattering studies, increased with increasing ILs concentration, which were due to the formation of larger aggregates; the aggregates are considered to be comprised of the anionic surfactant with a substantial proportion of ILs cation as the bound counter ions. Such studies are considered to shed further light in the fundamentals of IL induced micellization as well as in different practical applications.

The interfacial and micellization behavior of three dicarboxylic a mino acid-based a nionic s urfactants, abbreviated as AAS ( N -dodecyl derivative of -aminomalonate, -aspartate, and -glutamate) in combination with hexadecyltrimethylammonium bromide (HTAB) were investigated by surface tension, conductance, UV-vis absorption/emission spectroscopy, dynamic light scattering (DLS), and viscosity studies. Critical micelle concentration (CMC) values of the surfactant mixtures are significantly lower than the predicted values, indicating associative interaction between the components. Surface excess, limiting molecular area, surface pressure at the CMC, and Gibbs free energy indicate spontaneity of the micellization processes compared to the pure components. CMC values were also determined from the sigmoidal variation in the plot of micellar polarity and pyrene UV-vis absorption/emission intensities with surfactant concentration. The aggregation number, determined by static fluorescence quenching method, increases with decreasing mole fraction of the AAS (α AAS ), where the micelles are mainly dominated by the HTAB molecules. The size of the micelle increases with decreasing α AAS , leading to the formation of larger and complex aggregates, as also supported by the viscosity studies. Micelles comprising 20-40 mol % AAS are highly viscous, in consonance with their sizes. Some of the mixed surfactant systems show unusual viscosity (shear thickening and increased viscosity with increasing temperature). Such mixed surfactant systems are considered to have potential in gel-based drug delivery and nanoparticle synthesis.

Arsenic created a serious public health problem in Bangladesh due to its presence in groundwater and dissemination of the toxic effects to millions of people. The scarcity of the treatment options to manage this affected population has made the situation much worse. To find a promising treatment option, this study was undertaken to examine the ameliorating roles of Syzygium cumini leaf extract (SLE) against arsenic-induced toxic effects in mice. Swiss albino mice were divided into four groups where 'control' group received pure water + normal feed, 'arsenic (As)' group received sodium arsenite (NaAsO 2 )-containing water (10 μg/g body weight/day) + normal feed, 'As+SLE' group received NaAsO 2 -containing water + feed supplemented with SLE (50 µg/g body weight/day) and finally the 'SLE' group received pure water + feed supplemented with SLE. A gradual increase in body weight gain was observed in control mice; however, the body weight gain in As-exposed mice was decreased. This decrease in body weight gain was prevented in As+SLE group mice that received SLE supplemented feed. Arsenic showed a secondary effect by causing enlargement of spleen, kidney and liver of 'As' group mice and this enlargement of the organs was minimized with SLE supplementation. In addition, SLE abrogated arsenic-mediated elevation of serum alkaline phosphatase (ALP), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), uric acid and glucose. These results, therefore, suggest that SLE might have future therapeutic value for preventing or reducing arsenic-induced toxic effects.