A highly sensitive fluorescence biosensor for aflatoxins B1 detection based on polydiacetylene liposomes combined with exonuclease III-assisted recycling amplification.

A fluorescence biosensor for determination of aflatoxin B1 (AFB1) based on polydiacetylene (PDA) liposomes and exonuclease III (EXO III)-assisted recycling amplification was developed. The AFB1 aptamer partially hybridizes with complementary DNA (cDNA), which is released upon recognition of AFB1 by the aptamer. Subsequently, the cDNA hybridizes with hairpin H to form double-stranded DNA that undergoes digestion by EXO III, resulting in the cyclic release of cDNA and generation of capture DNA for further reaction. The capture DNA then hybridizes with probe modified on PDA liposomes, leading to aggregation of liposomes and subsequent fluorescence production. This strategy exhibited a limit of detection of 0.18 ng/mL within the linear range 1-100 ng/mL with a determination coefficient > 0.99. The recovery ranged from 92.81 to 106.45%, with relative standard deviations (RSD) between 1.73 and 4.26%, for corn, brown rice, peanut butter, and wheat samples. The stability, accuracy, and specificity of the method demonstrated the applicability for real sample analysis.

Conjugation Length-Dependent Raman Scattering Intensity of Conjugated Polymers.

Polydiacetylenes, as a class of conjugated polymers with alternating conjugated C═C and C≡C bonds, have emerged as a promising probe material for biomedical Raman imaging, given their ultrastrong Raman scattering intensity. However, the relationship between the structure, especially the molecular length of polydiacetylenes, and their Raman scattering intensity remains unclear. In this work, a series of water-soluble polydiacetylenes, namely poly(deca-4,6-diynedioic acid) (PDDA) with different molecular weights (MWs), is prepared through controlled polymerization and degradation. The ultraviolet-visible (UV-vis) absorption spectroscopic and Raman spectroscopic studies on these polymers reveal that the Raman scattering intensity of PDDA increases nonlinearly with the MW. The MW-Raman scattering intensity relationship in the polymerization process is completely different from that in the degradation process. In contrast, the Raman scattering intensity increases more linearly with the maximal absorbance of the polymer, and the relationship between the Raman scattering intensity and the maximal absorbance of PDDA in the polymerization process is consistent with that in the degradation process. The Raman scattering intensity of PDDA hence exhibits a better dependence on the effective conjugation length of the polymer, which should guide the future design of conjugated polymers for Raman imaging applications.

Colorimetric detection of alkaline phosphatase activity based on pyridoxal phosphate-induced chromatic switch of polydiacetylene nano-liposomes.

A colorimetric assay based on polydiacetylenes (PDA) nano-liposomes is reported for facile and sensitive detection of alkaline phosphatase (ALP) activity. The critical basis of this method is that the interaction of pyridoxal phosphate (PLP) with nitrogenous group functionalized PDA nano-liposomes induces distinct blue-to-red color changes of PDA nano-liposomes. In the presence of ALP, as a nature substrate, PLP is enzymatically hydrolyzed to form pyridoxal, which cannot interact with PDA nano-liposomes. As a result, the concentration of PLP is reduced and the color change of PDA nano-liposomes is retarded, which is associated with ALP level. Under optimal conditions, the proposed method showed good linear relationship with ALP activity in the range 10-200 U/L with a limit of detection of 2.8 U/L. The detection process could be vividly observed with the naked eye. Additional attempts by using the method for the evaluation of inhibitor efficiency were also achieved with satisfying results. The method was further challenged with real human serum samples, showing consistent results when compared with a commercial standard assay kit. Such simple and easy-to-use approach may provide a new alternative for clinical and biological detection of ALP.

UV-guided isolation of enantiomeric polyacetylenes from Bupleurum scorzonerifolium Willd. with inhibitory effects against LPS-induced NO release in BV-2 microglial cells.

UV-guided fractionation led to the isolation of thirteen new polyacetylenes (1-13) from the roots of Bupleurum scorzonerifolium Willd. All polyacetylenes were analyzed as racemates since the lack of optical activity and Cotton effects in the ECD spectra. The sequent chiral-phase HPLC resolution successfully gave twelve pairs of enantiomers 1a/1b and 3a/3b-13a/13b. Their structures were elucidated based on the HRESIMS and NMR data analyses. The absolute configurations were determined by the combination of Snatzke's method, electronic circular dichroism calculations, and single-crystal X-ray diffraction. Using Griess methods and MTT assays, polyacetylenes 1a, 3a, 4a/4b-12a/12b, and 13a displayed inhibitory activities against LPS-induced NO release in BV-2 microglial cells.

Merging Supramolecular and Covalent Helical Polymers: Four Helices Within a Single Scaffold.

Supramolecular and covalent polymers share multiple structural effects such as chiral amplification, helical inversion, sergeants and soldiers, or majority rules, among others. These features are related to the axial helical structure found in both types of materials, which are responsible for their properties. Herein a novel material combining information and characteristics from both fields of helical polymers, supramolecular (oligo(p-phenyleneethynylene) (OPE)) and covalent (poly(acetylene) (PA)), is presented. To achieve this goal, the poly(acetylene) must adopt a dihedral angle between conjugated double bonds (ω1) higher than 165°. In such cases, the tilting degree (Θ) between the OPE units used as pendant groups is close to 11°, like that observed in supramolecular helical arrays of these molecules. Polymerization of oligo[(p-phenyleneethynylene)n]phenylacetylene monomers (n = 1, 2) bearing L-decyl alaninate as the pendant group yielded the desired scaffolds. These polymers adopt a stretched and almost planar polyene helix, where the OPE units are arranged describing a helical structure. As a result, a novel multihelix material was prepared, the ECD spectra of which are dominated by the OPE axial array.

Polydiacetylene vesicles acting as colorimetric sensor for the detection of plantaricin LD1.

The interaction of antimicrobial peptides with membrane lipids plays a major role in numerous physiological processes. In this study, polydiacetylene (PDA) vesicles were synthesized using 10, 12-tricosadiynoic acid (TRCDA) and 1, 2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC). These vesicles were applied as artificial membrane biosensor for the detection of plantaricin LD1 purified from Lactobacillus plantarum LD1. Plantaricin LD1 (200 µg/mL) was able to interact with PDA vesicles by changing the color from blue to red with colorimetric response 30.26 ± 0.59. Nisin (200 µg/mL), used as control, also changed the color of the vesicles with CR% 50.56 ± 0.98 validating the assay. The vesicles treated with nisin and plantaricin LD1 showed increased infrared absorbance at 1411.46 and 1000-1150 cm-1 indicated the interaction of bacteriocins with phospholipids and fatty acids, respectively suggesting membrane-acting nature of these bacteriocins. Further, microscopic observation of bacteriocin-treated vesicles showed several damages indicating the interaction of bacteriocins. These findings suggest that the PDA vesicles may be used as bio-mimetic sensor for the detection of bacteriocins produced by several probiotics in food and therapeutic applications.

Secondary Metabolites from the Marine Sponges of the Genus Petrosia: A Literature Review of 43 Years of Research.

Sponges are prolific sources of various natural products that have provided the chemical scaffolds for new drugs. The sponges of the genus Petrosia inhabit various regions and contain a variety of biologically active natural products such as polyacetylenes, sterols, meroterpenoids, and alkaloids. This review aims to provide a comprehensive summary of the chemical structures and biological activities of Petrosia metabolites covering a period of more than four decades (between 1978 and 2020). It is also described in this review that the major groups of metabolites from members of the genus Petrosia differed with latitude. The polyacetylenes were identified to be the most predominant metabolites in Petrosia sponges in temperate regions, while tropical Petrosia species were sources of a greater variety of metabolites, such as meroterpenoids, sterols, polyacetylenes, and alkaloids.

PDA-Based Glyconanomicelles for Hepatocellular Carcinoma Cells Active Targeting Via Mannose and Asialoglycoprotein Receptors.

Hepatocellular carcinoma (HCC) is the sixth most common neoplasia and the fourth most common cause of cancer-related mortality worldwide. Sorafenib is the first-line molecular therapy for patients in an advanced stage of HCC. However, the recommended clinical dose of Sorafenib is associated with several complications, which derive from its lack of cell specificity and its very low water solubility. To circumvent these drawbacks, in the present study we developed two sugar-coated polydiacetylene-based nanomicelles-Sorafenib carriers targeting mannose and asialoglycoprotein receptors (MR and ASGPR, respectively). The strategies allowed the inducement of apoptosis and reduction of cell proliferation at a nanomolar, instead of micromolar, range in liver cancer cells. The study showed that, contrary to literature data, Sorafenib included into the pMicMan (Man = mannose) vector (targeting MR) is more efficient than pMicGal (Gal = galactose) (targeting ASGPR). Indeed, pMicMan increased the endosomal incorporation with an increased intracellular Sorafenib concentration that induced apoptosis and reduced cell proliferation at a low concentration range (10-20 nM).

Kamiohnoyneosides A and B, two new polyacetylene glycosides from flowers of edible Chrysanthemum "Kamiohno".

Two new polyacetylene glycosides, kamiohnoyneosides A and B, were isolated from the flowers of edible Chrysanthemum "Kamiohno", along with a known polyacetylene glycoside and two known monoterpene glycosides. The structures of new compounds were elucidated on the basis of spectroscopic data. Kamiohnoyneoside A and three known compounds moderately inhibited formation of Nε-(carboxymethyl)lysine, one of the representative advanced glycation endproducts.

Osirisynes G-I, New Long-Chain Highly Oxygenated Polyacetylenes from the Mayotte Marine Sponge Haliclona sp.

Chemical study of the CH2Cl2-MeOH (1:1) extract from the sponge Haliclona sp. collected in Mayotte highlighted three new long-chain highly oxygenated polyacetylenes, osirisynes G-I (1-3) together with the known osirisynes A (4), B (5), and E (6). Their structures were elucidated by 1D and 2D NMR spectra and HRESIMS and MS/MS data. All compounds were evaluated on catalase and sirtuin 1 activation and on CDK7, proteasome, Fyn kinase, tyrosinase, and elastase inhibition. Five compounds (1; 3-6) inhibited proteasome kinase and two compounds (5-6) inhibited CDK7 and Fyn kinase. Osirisyne B (5) was the most active compound with IC50 on FYNB kinase, CDK7 kinase, and proteasome inhibition of 18.44 µM, 9.13 µM, and 0.26 µM, respectively.

Development and Evaluation of the Chromatic Behavior of an Intelligent Packaging Material Based on Cellulose Acetate Incorporated with Polydiacetylene for an Efficient Packaging.

Global growth of the food industry and the demand for new products with natural characteristics, safe conditions and traceability have driven researches for the development of technologies such as intelligent packaging, capable to fulfil those needs. Polydiacetylene (PDA) is a synthetic material that has been highlighted in research field as a sensor substance, which can be used to produce intelligent packaging capable to detect chemical or biochemical changes in foods and in their environment due to PDA's color transition from blue to red. This work focused on the development and optimization of an intelligent packaging constituted of a polymeric matrix of cellulose acetate-based incorporated with PDA as the substance sensor. Cellulose acetate films (3% wt.) were developed by a casting method, and the amounts of triethyl citrate plasticizer (TEC) (0-25% wt. of cellulose-acetate) and PDA (0-60 mg) were analyzed to optimize the conditions for the best color transitioning at this study range. The compound amounts incorporated into polymeric matrices were established according to Central Composite Designs (CCD). Three more design variables were analyzed, such as the polymerization time of PDA under UV light exposition (0-60 min), pH values (4-11) and temperature exposure on the film (0-100 °C), important factors on the behavior of PDA's color changing. In this study, film thickness and film color coordinates were measured in order to study the homogeneity and the color transitioning of PDA films under different pH and temperature conditions, with the purpose of maximizing the color changes through the optimization of PDA and TEC concentrations into the cellulose acetate matrix and the polymerization degree trigged by UV light irradiation. The optimal film conditions were obtained by adding 50.48 g of PDA and 10% of TEC, polymerization time of 18 min under UV light, at 100 °C ± 2 °C of temperature exposure. The changes in pH alone did not statistically influence the color coordinates measured at the analyzed ratio; however, variations in pH associated with other factors had a significant effect on visual color changes, and observations were described. PDA films were optimized to maximize color change in order to obtain a cheap and simple technology to produce intelligent packaging capable to monitor food products along the distribution chain in real time, improving the food quality control and consumer safety.

A polydiacetylene-based colorimetric sensor as an active use-by date indicator for milk.

Current food date marking tools fail to take into account variables that impact shelf life, leading to food waste and posing human health risks. Herein, we develop a polydiacetylene/zinc oxide (PDA/ZnO) colorimetric sensor that can indicate milk freshness in real time. The PDA/ZnO system is engineered to indicate the freshness of full cream milk by monitoring its pH based on lactic acid concentration, an indicator of bacterial spoilage. The sensitivity is tuned by selection of the diacetylene monomers from which it is composed. Designs constructed from shorter chain diacetylenes are found to undergo chromatic transitions at a lower lactic acid concentration. Two new methods to tune PDA/ZnO sensitivity to lactic acid are explored, including relocation of the diacetylene functional group and lipid doping. A system constructed from 5,7-hexadecadiynoic acid (HDDA) and ZnO can discriminate between fresh (pH 6.8-6.0), spoiling (pH 6.0-4.5), and spoiled milk (pH 4.5-4.0) by a respective blue to purple to red color change. The chromatic stability of the PDA/ZnO sensor in food is studied as previous reports have shown the system to be unstable. We present a new method that stabilizes the nanocomposite by pre-exposure to the food matrix prior to photopolymerization.

Development of color changing polydiacetylene-based biomimetic nanovesicle platforms for quick detection of membrane permeability across the blood brain barrier.

Membrane permeability through passive diffusion is one of the important pathways for passage of drugs across the blood brain barrier (BBB). The present study describes the development of biomimetic unilamellar lipopolymeric nanovesicles of size 268 ± 37 nm, consisting of polar brain lipids in conjunction with polydiacetylene and validation of their application for an abbreviated yet accurate membrane permeability assay with high-throughput and rapid identification of BBB permeability of drugs. The nanovesicle suspension was tested with drugs of known permeability across the BBB to validate the detection of changes in hue, absorbance and fluorescence in response to permeation across the nanovesicles. A simple device was developed based on the nanovesicle sensors along with a mobile application which allowed for the determination of hue corresponding to qualitative identification of whether a drug is BBB permeable (BBB+) or not (BBB-). With respect to determination of a suitable endpoint in this assay, a hue cut off of 275°, reduction in %blueness by less than 59% and a fluorescence intensity of ≥0.22 a.u. at 560 nm accurately differentiated between drugs which are permeable and impermeable across the BBB within 5 minutes. Further quantification of BBB permeability can be done through the concentration at which the above end-points are achieved. For the quantification of the permeability, absorbance and fluorescence measurements were performed. The device thus developed allows the rapid determination of BBB permeability of various agents in drug discovery especially in smaller set-ups with minimal equipment through changes in color, absorbance and fluorescence.

Rapid evaluation of gold nanoparticle-lipid membrane interactions using a lipid/polydiacetylene vesicle sensor.

Surface modification of gold nanoparticles (AuNPs) has significant and complicated effects on their interactions with cell membranes. In this study, we used a lipid/polyacetylene (PDA) vesicle sensor as the lipid membrane model to evaluate AuNP-lipid membrane interactions. Based on the colorimetric response (CR) of PDA vesicles before and after incubation with AuNPs, it was found that the interaction was highly dependent on the surface charge of AuNPs. As compared to the positively charged NPs, neutral and zwitterionic NPs adsorbed much less on the lipid membrane. Negatively charged NPs did not induce any noticeable color changes even at high concentrations. A class of cationic AuNPs with different degrees of surface hydrophobicity was further selected to investigate the role of hydrophobicity in interacting with lipid/PDA vesicles, and log(EC50) was employed as the evaluation index. According to the log(EC50)-NP concentration curve, the hydrophobicity of NPs enhanced the lipid membrane affinity, but electrostatic interactions weakened this effect. Finally, different concentrations of bovine serum albumin (BSA) were used to study the effect of the protein corona on NP-lipid membrane interactions. The formation of a NP-protein corona was found to mask the electrostatic interactions, leading to the decrease of the CR values of cationic NPs, and highly hydrophobic NPs were less affected by a low concentration of BSA due to the strong hydrophobic interactions. Although the effect of NP surface properties on their interactions with cells is far more complicated, our study provides a rapid and effective method for the evaluation of the interactions between surface modified AuNPs and lipid membranes.

Nanotoxicology at the particle/micelle frontier: influence of core-polymerization on the intracellular distribution, cytotoxicity and genotoxicity of polydiacetylene micelles.

The understanding of the cellular uptake and the intracellular fate of nanoparticles and their subsequent influence on cell viability is challenging as far as micelles are concerned. Such systems are dynamic by nature, existing as unimers under their critical micelle concentration (CMC), and as micelles in equilibrium with unimers above the CMC, making canonical dose-response relationships difficult to establish. The purpose of this study was to investigate the in vitro cytotoxicity and uptake of two micellar sytems that are relevant for drug delivery. The two micelles incorporate a poly(ethylene glycol) coating and a pentacosadiynoic core which is either polymerized (pDA-PEG micelles) or non-polymerized (DA-PEG micelles), with the aim of evaluating the influence of the micelles status ("particle-like" or "dynamic", respectively) on their toxicological profile. Intracellular distribution and cytotoxicity of polymerized and non-polymerized micelles were investigated on RAW 264.7 macrophages in order to compare any different interactions with cells. Non-polymerized micelles showed significantly higher cytotoxicity than polymerized micelles, especially in terms of cell permeabilization, correlated to a higher accumulation in cell membranes. Other potential toxicity endpoints of polymerized micelles were then thoroughly studied in order to assess possible responses resulting from their endocytosis. No specific mechanisms of cytotoxicity were observed, neither in terms of apoptosis induction, cell membrane damage, release of inflammatory mediators nor genotoxicity. These data indicate that non-polymerized micelles accumulate in the cell membrane and induce cell membrane permeabilization, resulting in significant toxicity, whereas polymerized, stable micelles are internalized by cells but exert no or very low toxicity.

Polydiacetylene-based ultrastrong bioorthogonal Raman probes for targeted live-cell Raman imaging.

Live-cell Raman imaging based on bioorthogonal Raman probes with distinct signals in the cellular Raman-silent region (1800-2800 cm-1) has attracted great interest in recent years. We report here a class of water-soluble and biocompatible polydiacetylenes with intrinsic ultrastrong alkyne Raman signals that locate in this region for organelle-targeting live-cell Raman imaging. Using a host-guest topochemical polymerization strategy, we have synthesized a water-soluble and functionalizable master polydiacetylene, namely poly(deca-4,6-diynedioic acid) (PDDA), which possesses significantly enhanced (up to ~104 fold) alkyne vibration compared to conventional alkyne Raman probes. In addition, PDDA can be used as a general platform for multi-functional ultrastrong Raman probes. We achieve high quality live-cell stimulated Raman scattering imaging on the basis of modified PDDA. The polydiacetylene-based Raman probes represent ultrastrong intrinsic Raman imaging agents in the Raman-silent region (without any Raman enhancer), and the flexible functionalization of this material holds great promise for its potential diverse applications.

Enantioselective Synthesis and Antifungal Activity of C18 Polyacetylenes.

Fungal pathogens cause serious crop diseases and decrease crop yields and quality. Polyacetylene alcohols are plant secondary metabolites and bioactive against various pathogenic fungi. They are, however, difficult to synthesize. In the present study, an efficient and highly enantioselective method (>98% ee) was established and employed to achieve the synthesis of the natural C18 polyacetylenes (S,E)-octadeca-1,9-dien-4,6-diyn-3-ol 1, (3R,10R,E)-octadeca-1,8-dien-4,6-diyne-3,10-diol 2, and their analogs. The title compounds were structurally characterized and biologically evaluated for fungicidal activities. The compounds exhibited high potencies against eight pathogenic fungal species tested, such as Colletotrichum gloeosporioiles, Bipolaris sorokiniana, Fusarium graminearum, and Fusarium pseudograminearum, with half-maximum effective concentrations ranging from 8 to 425 µg/mL, being similar to those of the fungicide thiophanate-methyl (3-408 µg/mL). These compounds are potential natural fungicides and fungicide lead candidates for further structural and property improvements.

Characterization of novel polydiacetylene gel dosimeter for radiotherapy.

Polymer gel dosimeters are instrumental for clinical and research applications in radiotherapy. These dosimeters possess the unique ability to record dose distribution in three dimensions. A Polymer gel dosimeter is composed of organic molecules in a gel matrix, which upon irradiation polymerize to form a conjugated polymer with optical absorbance proportional to the irradiated dose. Other required characteristics of a radiotherapy clinical dosimeter are soft-tissue equivalency, linear dose-response in a range of clinical treatments, and long term stability for the duration of the analysis. The dosimeter presented in this paper is based on diacetylene bearing fatty acid aggregates embedded in a soft-tissue equivalent gel matrix, Phytagel™, which upon irradiation polymerize to form a blue phase polydiacetylene with a strong optical absorption. Initial characterization showed that PDA-gel irradiated with 160 kV x-ray responded linearly to the irradiated dose, and the calculated diffusion coefficient is [Formula: see text] what is very low. It was also found that the percentage depth dose (PDD) curve of the PDA-gel in a 4 × 4 cm2 field, irradiated with 6 MV x-rays, was with good agreement with the literature. PDA-gel has the potential to detect absorbed dose in a range of clinical radiological irradiation regimes.

Isolation and structure elucidation of two new compounds from artemisia Ordosica Krasch.

Two new compounds, namely arteordoyn A (1) and arteordoyn B (2), together with four known compounds, were isolated from the petroleum ether extract of Artemisia ordosica Krasch. The structures elucidation of 1 and 2 were carried out by 1D-NMR (1H and 13C NMR), 2D-NMR (COSY, HSQC, HMBC and NOESY) and HR-ESI-MS spectral analysis.

New cytotoxic polyacetylene alcohols from the Egyptian marine sponge Siphonochalina siphonella.

Three new polyacetylenic alcohols, siphonellanols A-C (1-3), together with two known polyacetylenic alcohols (4-5), were isolated from the CHCl3-soluble fraction of the methanolic extract of the marine sponge Siphonochalina siphonella, collected in Egypt. The structures of 1-3 were determined by spectroscopic analyses of their 1D-, 2D-NMR, and MS spectra and by comparisons with reported data. The cytotoxicity assay revealed that 1-3 exhibited moderate cytotoxic activities against a human cervical cancer cell line (HeLa), a human breast cancer cell line (MCF-7), and a human lung cancer cell line (A549) with IC50 values ranging from 25.9 to 69.2 µM.