Antimicrobial peptide mimetic minimalistic approach leads to very short peptide amphiphiles-gold nanostructures for potent antibacterial activity.

Strategically controlling concentrations of lipid-conjugated L-tryptophan (vsPA) guides the self-assembly of nanostructures, transitioning from nanorods to fibres and culminating in spherical shapes. The resulting Peptide-Au hybrids, exhibiting size-controlled 1D, 2D, and 3D nanostructures, show potential in antibacterial applications. Their high biocompatibility, favourable surface area-to-volume ratio, and plasmonic properties contribute to their effectiveness against clinically relevant bacteria. This controlled approach not only yields diverse nanostructures but also holds promise for applications in antibacterial therapeutics.

Amyloidogenic Propensity of Metabolites in the Uric Acid Pathway and Urea Cycle Critically Impacts the Etiology of Metabolic Disorders.

Novel insights into the etiology of metabolic disorders have recently been uncovered through the study of metabolite amyloids. In particular, inborn errors of metabolism (IEMs), including gout, Lesch-Nyhan syndrome (LNS), xanthinuria, citrullinemia, and hyperornithinemia-hyperammonemia-homocitrullinuria (HHH) syndrome, are attributed to the dysfunction of the urea cycle and uric acid pathway. In this study, we endeavored to understand and mechanistically characterize the aggregative property exhibited by the principal metabolites of the urea cycle and uric acid pathway, specifically hypoxanthine, xanthine, citrulline, and ornithine. Employing scanning electron microscopy (SEM), transmission electron microscopy (TEM), and atomic force microscopy (AFM), we studied the aggregation profiles of the metabolites. Insights obtained through molecular dynamics (MD) simulation underscore the vital roles of π-π stacking and hydrogen bonding interactions in the self-assembly process, and thioflavin T (ThT) assays further corroborate the amyloid nature of these metabolites. The in vitro MTT assay revealed the cytotoxic trait of these assemblies, a finding that was substantiated by in vivo assays employing the Caenorhabditis elegans (C. elegans) model, which revealed that the toxic effects were more pronounced and dose-specific in the case of metabolites that had aged via longer preincubation. We hence report a compelling phenomenon wherein these metabolites not only aggregate but transform into a soft, ordered assembly over time, eventually crystallizing upon extended incubation, leading to pathological implications. Our study suggests that the amyloidogenic nature of the involved metabolites could be a common etiological link in IEMs, potentially providing a unified perspective to study their pathophysiology, thus offering exciting insights into the development of targeted interventions for these metabolic disorders.

An Isothiazolanthrone-Based Self-Assembling Anticancer Color-Changing Dye for Concurrent Imaging and Monitoring of Cell Viability.

We report the photophysical properties, self-assembly and biological evaluation of an isothiazolanthrone-based dye, 7-amino-6H-anthra[9,1-cd]isothiazol-6-one (AAT), which reveals anticancer properties and can be potentially used as dye for monitoring cell viability. The solvent-dependent photophysical studies suggest that the emission of AAT is sensitive to environment polarity due to which interesting changes in the colored emission may be observed owing to the charge transfer (CT) processes. AAT also self-assembles to tree-like branched morphologies and produce, a greenish emission inside the cells when imaged after short interval (15 mins) of incubation while a red fluorescence could be noted after 24 h. Interestingly, AAT also produce differential emission inside mouse normal cells as compared to its cancer cell lines since it possess anticancer activity. The experimental observations were also validated theoretically via computational modeling.

Antimicrobial nano-assemblies of tryptocidine C, a tryptophan-rich cyclic decapeptide, from ethanolic solutions.

Tryptocidine C (TpcC), a Trp-rich cyclodecapeptide is a minor constituent in the antibiotic tyrothricin complex from Brevibacillus parabrevis. TpcC possesses a high tendency to oligomerise in aqueous solutions and dried TpcC forms distinct self-assembled nanoparticles. High-resolution scanning electron microscopy revealed the influence of different ethanol:water solvent systems on TpcC self-assembly, with the TpcC, dried from a high concentration in 15% ethanol, primarily assembling into small nanospheres with 24.3 nm diameter and 0.05 polydispersity. TpcC at 16 µM, near its CMC, formed a variety of structures such as small nanospheres, large dense nanospheroids and facetted 3-D-crystals, as well as sheets and coarse carpet-like structures which depended on ethanol concentration. Drying 16 µM TpcC from 75% ethanol resulted in highly facetted 3-D crystals, as well as small nanospheres, while those in 10% ethanol preparation had less defined facets. Drying from 20 to 50% ethanol led to polymorphic architectures with a few defined nanospheroids and various small nanoparticles, imbedded in carpet- and sheet-like structures. These polymorphic surface morphologies correlated with maintenance of fluorescence properties and the surface-derived antibacterial activity against Staphylococcus aureus over time, while there was a significant change in fluorescence and loss in activity in the 10% and 75% preparations where 3-D crystals were observed. This indicated that TpcC oligomerisation in solutions with 20-50% ethanol leads to metastable structures with a high propensity for release of antimicrobial moieties, while those leading to crystallisation limit active moieties release. TpcC nano-assemblies can find application in antimicrobial coatings, surface disinfectants, food packaging and wound healing materials.

Short peptide amphiphile nanostructures facilitate sunlight-induced nanowelding of gold nanosheets.

An effortless thermoplasmonic welding of multi-shaped gold nanosheets is achieved by ordinary and simple sunlight irradiation. A light-matter interaction occurred via the nanogaps of smaller nanosheets, leading to the enhancement of the electromagnetic field and thus effectively concentrating the heat at the welding point. The sPA peptide nanostructure facilitates the nanowelding of small caged gold nanostructures.

Mercury-instructed assembly (MiA): architecting clathrin triskelion-inspired highly functional C3-symmetric triskelion nanotorus functional structures into microtorus structures.

To detect heavy metal toxicity using self-assembled nanostructures, a clathrin triskelion-inspired highly functional C3-symmetric trimerized biotinylated di-tryptophan peptide was used. This triskelion peptide is known to self-assemble into nanotorus-like structures and can therefore act as a nanocage for various analytes. In this work, in addition to spectroscopy, force and electron microscopy were successfully used to detect the effect of toxic metal ions such as zinc, cadmium, and mercury by exploiting the change in the nanotorus morphology. Different concentrations of mercury led to the expansion of nanotorus structures into microtori. Therefore, we provide a unique application of heavy metal toxicity by utilizing "material nanoarchitectonics" to architect nanotorus structures into higher-order microtorus structures, as instructed by mercury. Such a strategy can make heavy metal sensing easier for materials scientists and open new avenues for biomedical/environmental science applications.

A new aggregation induced emission enhancement (AIEE) dye which self-assembles to panchromatic fluorescent flowers and has application in sensing dichromate ions.

We report for the very first time the crystal structure and self-assembly of a new aggregation-induced emission enhancement (AIEE) dye 4-(5-methoxythiazolo[4,5-b]pyridin-2-yl)-N,N-dimethylaniline (TPA) and its application in sensing dichromate ions. TPA reveals cyan blue emission under UV and visible light. The self-assembly properties of TPA were studied extensively by scanning electron microscopy (SEM) which revealed the formation of beautiful flower-like morphologies. These structures revealed both green and red fluorescence under FITC and rhodamine filters respectively when observed through fluorescence microscopy connoting the panchromatic emission properties of TPA from blue to red. The interactions which cause self-assembled structure formation in TPA were also validated theoretically using density functional theory (DFT) calculations. Crystal and molecular structure analysis of TPA was carried out via single-crystal X-ray diffraction to visualize the intermolecular interactions occurring in the solid-state and to study the structure-photophysical property relationship in the aggregated state. The photophysical properties of TPA were also studied extensively by UV-visible and fluorescence spectroscopy and its quantum yield and fluorescence lifetime were calculated by time-correlated single-photon counting (TCSPC). Interestingly, TPA could efficiently sense dichromate (Cr2O72-) ions in an acidic medium and an interesting morphological transition from a fluorescent flower to non-fluorescent disassembled structures could also be observed. The limit of detection of TPA for Cr2O72- ions was found to be as low as 5.5 nM, suggesting its exceptional sensitivity. More importantly, TPA could selectively sense Cr2O72- ions in real water samples even in the presence of other metal ions routinely present in polluted water, hence making it practically useful for water quality monitoring.

Microalgal drugs: A promising therapeutic reserve for the future.

Over the decades, a variety of chemically synthesized drugs are being used to cure existing diseases but often these drugs could not be effectively employed for the treatment of serious and newly emerging diseases. Fortunately, in nature there occurs immense treasure of plants and microorganisms which are living jewels with respect to their richness of medically important metabolites of high value. Hence, amongst the existing microorganism(s), the marine world offers a plethora of biological entities that can contribute to alleviate numerous human ailments. Algae are one such photosynthetic microorganism found in both marine as well as fresh water which are rich source of metabolites known for their nutrient content and health benefits. Various algal species like Haematococcus, Diatoms, Griffithsia, Chlorella, Spirulina, Ulva, etc. have been identified and isolated to produce biologically active and pharmaceutically important high value compounds like astaxanthin, fucoxanthin, sulphur polysaccharides mainly galactose, rhamnose, xylose, fucose etc., which show antimicrobial, antifungal, anti-cancer, and antiviral activities. However, the production of either of these bio compounds is favored under conditions of stress. This review gives detailed information on various nutraceutical metabolites extracted from algae. Additionally focus has been made on the role of these bio compounds extracted from algae especially sulphur polysaccharides to treat several diseases with prospective treatment for SARS-CoV-2. Lastly it covers the knowledge gaps and future perspectives in this area of research.

Unusual Aggregates Formed by the Self-Assembly of Proline, Hydroxyproline, and Lysine.

There is a plethora of significant research that illustrates toxic self-assemblies formed by the aggregation of single amino acids, such as phenylalanine, tyrosine, tryptophan, cysteine, and methionine, and their implication on the etiology of inborn errors of metabolisms (IEMs), such as phenylketonuria, tyrosinemia, hypertryptophanemia, cystinuria, and hypermethioninemia, respectively. Hence, studying the aggregation behavior of single amino acids is very crucial from the chemical neuroscience perspective to understanding the common etiology between single amino acid metabolite disorders and amyloid diseases like Alzheimer's and Parkinson's. Herein we report the aggregation properties of nonaromatic single amino acids l-proline (Pro), l-hydroxyproline (Hyp), and l-lysine hydrochloride (Lys). The morphologies of the self-assembled structures formed by Pro, Hyp, and Lys were extensively studied by various microscopic techniques, and controlled morphological transitions were observed under varied concentrations and aging times. The mechanism of structure formation was deciphered by concentration-dependent 1H NMR analysis, which revealed the crucial role of hydrogen bonding and hydrophobic interactions in the structure formation of Pro, Hyp, and Lys. MTT assays on neural (SHSY5Y) cell lines revealed that aggregates formed by Pro, Hyp, and Lys reduced cell viability in a dose-dependent manner. These results may have important implications in the understanding of the patho-physiology of disorders such as hyperprolinemia, hyperhydroxyprolinemia, and hyperlysinemia since all these IEMs are associated with severe neurodegenerative symptoms, including intellectual disability, seizures, and psychiatric problems. Our future studies will endeavor to study these biomolecular assemblies in greater detail by immuno-histochemical analysis and advanced biophysical assays.

A naphthalimide-based peptide conjugate for concurrent imaging and apoptosis induction in cancer cells by utilizing endogenous hydrogen sulfide.

The excessive production of endogenous hydrogen sulfide (H2S) in cancer cells leads to enhanced tumor growth and metastasis. On the other hand, decreased endogenous H2S suppresses tumor growth. The reported approaches for inhibiting tumor growth are selective silencing of the tumor-promoting genes and pharmacological inhibition of these proteins. To enhance the antitumor efficacy of frontline chemotherapeutic agents, herein, we synthesized a highly sensitive endogenous H2S responsive fluorescent probe, i.e., a hydrogen sulfide-sensing naphthalimide-based peptide conjugate (HSNPc), which showed selective inhibition of proliferation of cancer cells due to apoptosis induction. Furthermore, HSNPc suppressed the glycolytic reserve, a critical energy source for the proliferation of cancer cells. HSNPc also decreased the Young's modulus of HeLa cells compared to the control cells, which demonstrated a direct relation between cell apoptosis and cell stiffness. Taken together, we demonstrated the dual function of detection and killing of cancer cells by HSNPc that can be likened to a theranostic role.

Cultivation of diatom Pinnularia saprophila for lipid production: A comparison of methods for harvesting the lipid from the cells.

The present study underlines the application of centrifugal force and pulse electric field techniques along with its comparison to resonance energy to harvest lipid from a fixed number of Pinnularia saprophila cells. Sulpho phospho vanillin method for lipid, and analysis of cells via microscopy was done. It was found that a centrifugal force of 11110×g for 15 min allowed ~3.39% lipid to ooze out with 2.5% cell destruction. Alternatively, when same numbers of diatom cells were subjected to pulse electric field at 110 kV/27 mA for 10 µs, maximum lipid production of 2.86% with 21.19% cell death was observed. It was perceived that diatom cells in a micro resonating micro fluidic chamber for 20 min harvested 4.4% of lipid with 11.16% of cell death. However, microfluidic device needs to be scaled up using cheaper material instead of silicon wafer, to be an efficient technique to milk lipid from diatoms.

Exopolysaccharides directed embellishment of diatoms triggered on plastics and other marine litter.

In the present study, embellishment or beautification of diatoms on substrates like plastics, polydimethylsiloxane, graphite, glass plate, and titanium dioxide, triggered by exopolysaccharides was examined under laboratory conditions. Exopolysaccharides are secreted mainly by primary colonisers, bacteria, which is succeeded by secondary colonisers i.e. diatoms. Both diatom (Nitzschia sp.4) and bacteria (Bacillus subtilis) were exposed with substrates separately for 30 days. Diatoms adhere on substrates strongly, not only because of surface roughness of different substrates but also the nanoporous architecture of diatoms which enhanced their embellishment. This study attempted to identify the substrates that adhere to diatoms strongly and was mainly analyzed by scanning electron microscope and further the observations are well supported by math work software (MATLAB). The variation of diatom's binding on different substrates is due to the influence of marine litters on diatom population in ocean beds where they undergo slow degradation releasing macro, micro and nanoparticles besides radicals and ions causing cell death. Therefore a proof-of-concept model is developed to successfully deliver a message concerning benefit of using different diatom species.

Constitutionally Isomeric Aromatic Tripeptides: Self-Assembly and Metal-Ion-Modulated Transformations.

Self-assembling peptides based on aromatic amino acids can adopt diverse nanostructures which primarily depend on their molecular structures. Therefore, to understand the nature of self-assembly on the molecular level we rationally designed two constitutional isomers of short aromatic peptides. The first isomer consists of a tyrosine moiety at the N-terminus and the second isomer consists of a tyrosine moiety at the C-terminus of the FF peptide, a core recognition motif of Amyloid ß peptides. Therefore, it can be considered that both the designed tripeptides are the analogues of the FFF peptide with only atomic(-H) level replacement by -OH functional group on the first and last phenyl ring, respectively. The first isomer self-assembled into 2D porous nanosheets ("Nanowebs"), however the second isomers produced toroidal shapes with central spheres ("Nano-Saturn" like assemblies). Interestingly, the presence of the transition-metal ions (copper, zinc and iron) triggered the self-assembly of both the peptides into fibrous circular discs, nanomats and nanoplates like assembly.

Peptide Nanostructure-Mediated Antibiotic Delivery by Exploiting H2S-Rich Environment in Clinically Relevant Bacterial Cultures.

Stimuli-responsive self-destructing soft structures serve as versatile hosts for the encapsulation of guest molecules. A new paradigm for H2S-responsive structures, based on a modified tripeptide construct, is presented along with microscopy evidence of its time-dependent rupture. As a medicinally interesting application, we employed these commercial antibiotic-loaded soft structures for successful drug release and inhibition of clinically relevant, drug-susceptible, and methicillin-resistant Staphylococcus aureus.

Self-assembly of a Sequence-shuffled Short Peptide Amphiphile Triggered by Metal Ions into Terraced Nanodome-like Structures.

We highlight the structural diversity of strategically designed two short peptide amphiphiles (sPAs) and describe their structure-function relationship studies. The shuffling of two key amino acids, that is, tyrosine and phenylalanine, in a designed sPA lead to a pair of constitutional isomers. Such small and strategic alteration can bring a substantial change in the self-assembling pattern. Inspired from the naturally occurring metallopeptides, bioactive transition-metal ions were used for constructing the unusual nanostructures. Use of appropriate metal ions created bigger differences between the properties of these isomers and hence the self-assembly. Coordination of appropriate transition metal ions modifies the internal nanoscale structures of sPA, thus leading to the formation of vertically stacked terraced layers with decreasing size, which possess a high degree of dimensional regularity. We propose that such metal-induced terraced nanodome-like hierarchical self-assembly may have relevance for specific biotechnology applications.

Synthesis and Aggregation Studies of a Pyridothiazole-Based AIEE Probe and Its Application in Sensing Amyloid Fibrillation.

We report the aggregation and photophysical properties of a pyridothiazole-based, aggregation-induced, emission-enhancement (AIEE) luminogen 4-(5-methoxy-thiazolo[4,5-b]pyridin-2-yl)benzoic acid (PTC1) and its application for the sensitive detection and monitoring of amyloid fibrillation. The aggregation properties of the AIEE probe were extensively studied by atomic force microscopy (AFM) and dynamic light scattering (DLS), and it was noted that as aggregation increases the fluorescence of PTC1 also was increased. The fluorescence of PTC1 was quenched upon the addition of cupric (Cu2+) ions, while the fluorescence is regenerated in the presence of amyloid fibers. AFM studies reveal that the PTC1 molecules self-associate/aggregate to hairy micelle-like structures, which dissociate or disrupt in the presence of the Cu2+ ions and again reassemble in the presence of amyloid fibers. Hence, the quenching and regeneration of PTC1 fluorescence may be attributed to the disaggregation and aggregation-induced emission (AIE), respectively. Further, a comparative analysis of the performance of PTC1 was done with conventional Thioflavin T, which confirms it to be a more sensitive probe for the detection of the amyloid, both in the presence and absence of Cu2+ ions. The experimental results were also validated theoretically via molecular docking and simulation studies. Of note, a very simple, facile, and cost-effective methodology for the detection of the amyloid fibers is presented, wherein fluorescence quenching/enhancement can be visualized under the UV light without the use of sophisticated instrumentation techniques. The AIEE probe was designed using an unusual pyridothiazole scaffold unlike commonly used archetypal AIE scaffolds based on tetraphenylethene (TPE) and hexaphenylsilole (HPS). Hence, the work also has implications in designing future AIEE dyes based on the pyridothiazole scaffold reported.

TiO2 doped polydimethylsiloxane (PDMS) and Luffa cylindrica based photocatalytic nanosponge to absorb and desorb oil in diatom solar panels.

Our previous report(s) demonstrated that piezoelectric disc fabricated diatom solar panels worked as micro resonating devices. Such devices have potential to harvest oils from living diatom cultures. However, it is observed that the collection and separation of oil from culture media using these devices are found to be difficult due to the presence of both living and dead diatom cells, which simultaneously get collected during this process. In this study we made a highly biocompatible nanosponge using TiO2 nanoparticle doped polydimethylsiloxane (PDMS) and Luffa cylindrica. Such hybrid nanosponge selectively absorbs and desorbs oil on exposure to ultraviolet light. The fabricated PDMS-Luffa-TiO2 nanosponge was characterized by Fourier transform infrared spectroscopy, scanning electron microscopy and atomic force microscopy to show the surface characteristics and affinity of nanoparticles to the membranous structure of PDMS-Luffa. A maximum 38% oil absorption was found in PDMS-Luffa-TiO2 nanosponge which was almost double that of sponges made up of PDMS (19%), PDMS-Luffa (18%) and PDMS-TiO2 coated (24%). Thus PDMS-Luffa-TiO2 nanosponge serves as a selective and recyclable oil absorption membranous structure. Furthermore, this hybrid nanosponge exhibited excellent recyclability by repeated absorption-desorption processes on exposure to UV light.

Fast Fourier infrared spectroscopy to characterize the biochemical composition in diatoms.

Diatoms are photosynthetic unicellular microalgae and are nature's hidden source of several biosynthetic metabolites with their use in biofuel, food and drug industries. They mainly contain various lipids, sterols, isoprenoids and toxins with their use in apoptotic, fertility controlling and cancer drugs. Chemical studies on diatoms are limited due to various limitations such as variation of nutrients, contaminants and change in seasonal factors in the environment. To overcome these limitations, we obtained axenic cultures of 12 fresh-water diatom strains on the 22nd day of inoculation having a dry weight of 1 mg each and performed their Fourier transform infrared (FTIR) study for the detection of functional groups responsible for their chemical moiety. The spectral mapping showed a varied level of polyunsaturated fatty acids, amides, amines, ketone bodies and esters for their applications in various pharmacological, food and biofuel industries in the exponential phase of their growth in f/2 media. The FTIR study of the 12 diatom strains showed various similarities in the form of some common peak patterns ranging from 3000 to 3600 cm-1 for vO-H absorption. The symmetric stretching vibration frequency of Diadesmis confervaceae (V2) type species showed different behaviour than others in the spectral region starting from 1600 to 1700 cm-1. The absorption between 1500 and 1575 cm-1 reflects the presence of the -N-H group. Infrared (IR) absorptions falling between 1600 and 1700 cm-1 reflect the presence of amide's vC=O in all species. Placoneis elginensis (V8) type species showed an additional absorption band which is centred around 1735-1750 cm-1 which perhaps reflects the presence of ester's vC=O. Diadesmis confervaceae (V2), Nitzschia palea (V4), Placoneis elginensis (V8), Nitzschia palea var. debilis (V6), Nitzschia inconspicua (V10), Gomphonema parvulum (V11) and Sellaphora (V12) showed distinct structural features with important key functionalities that can make them essential drug markers in the pharmaceutical industry.

Ornamentation of Triskelion Peptide Nanotori to Produce Gold Nanoparticle (AuNP)-Embedded Peptide Nanobangles.

We report the formation of gold nanoparticle (AuNP)-biotinylated triskelion peptide hybrid nanostructures. These structures were created by using self-organization and mimicking strategies to develop AuNP-ornamented peptide nanobangles with predetermined properties on biocompatible surfaces. Such intelligent and fascinating nanomaterials can be used for several bio-nanotechnological applications.

Thermoplasmonic effect of silver nanoparticles modulates peptide amphiphile fiber into nanowreath-like assembly.

This study demonstrates the beneficial role of di-tryptophan containing short peptide amphiphiles (sPA), for the synthesis and stabilization of AgNPs in the presence of sunlight followed by garlanding of AgNPs along the fibrous network of sPA. Such hybrid structures were precisely and selectively moulded into a nanowreath-type morphology due to the thermoplasmonic effect of AgNPs, and can be used for several bio-nanotechnological applications.