Detailed Toxicity Studies Research Articles

Sulfated Alginate for Biomedical Applications.

Alginate (Alg) polymers have received much attention due to the mild conditions required for gel formation and their good bio-acceptability. However, due to limited interactions with cells, many drugs, and biomolecules, chemically modified alginates are of great interest. Sulfated alginate (S-Alg) is a promising heparin-mimetic that continues to be investigated both as a drug molecule and as a component of biomaterials. Herein, the S-Alg literature of the past five years (2017-2023) is reviewed. Several methods used to synthesize S-Alg are described, with a focus on new advances in characterization and stereoselectivity. Material fabrication is another focus and spans bulk materials, particles, scaffolds, coatings, and part of multicomponent biomaterials. The new application of S-Alg as an antitumor agent is highlighted together with studies evaluating safety and biodistribution. The high binding affinity of S-Alg for various drugs and heparin-binding proteins is exploited extensively in biomaterial design to tune the encapsulation and release of these agents and this aspect is covered in detail. Recommondations include publishing key material properties to allow reproducibility, careful selection of appropriate sulfation strategies, the use of cross-linking strategies other than ionic cross-linking for material fabrication, and more detailed toxicity and biodistribution studies to inform future work.

Appraisal of phytochemical constituents, antioxidant and antibacterial activities of folkloric Pentanema confertiflorum rootbark extracts

BackgroundMedicinal plants have cultural prominence in the management of ailments worldwide. The different organs of Pentanema confertiflorum (hereafter referred as P. confertiflorum) as have been used to ameliorate different diseases in Ethiopia. Thus, this study focused on the qualitative and quantitative phytochemical analysis, antioxidant, and antibacterial screening of P. confertiflorum rootbark solvent fractions. MethodsThe rootbark of P. confertiflorum was successively soaked with petroleum ether, chloroform, and methanol. The total phenolic (TPC), flavonoid (TFC), and alkaloid contents (TAC) of the different crude extracts were determined through Folin-Ciocalteu Reagent (FCR), aluminum chloride colorimetric, and bromocresol green (BCG) assays, respectively. In addition, 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging and ferric-reducing antioxidant power (FRAP) assays were employed to determine the antioxidant activity of crude extracts. Moreover, the antimicrobial activity of the crude extracts was assessed against different bacteria strains via the disc diffusion method. ResultsThe qualitative phytochemical investigation study resonated with the abundance of several secondary metabolites, especially in the methanolic rootbark extract (MRBE). In addition, the MRBE had the highest TPC (76.7 mg GAE/g extracts), TFC (86.89 mg QE/g extracts), and TAC (80.68 mg ATE/g extracts) as compared to petroleum ether and chloroform extracts. Consistent with its TPC, TFC, and TAC, the MRBE demonstrated better antioxidant activity against DPPH radical scavenging (IC50 = 44.53 ppm) and FRAP assays (EC50 = 42.72 ppm). Moreover, all the rootbark extracts induced dose-dependent antibacterial activity against Streptococci pyogenes and Staphylococcus aureus, Klebsiella pneumoniae, and Escherichia coli. ConclusionsThe preliminary findings suggested the presence of a strong correlation between the folkloric use of P. confertiflorum and its observed antioxidant and antibacterial activity. Rigorous studies on the extracts such as the isolation and characterization of bioactive compounds, and detailed bioactivity and toxicity studies are desirable to bring about safe and effective lead compounds.

Biofunctionalized CdS Quantum Dots: A Case Study on Nanomaterial Toxicity in the Photocatalytic Wastewater Treatment Process.

The toxic nature of inorganic nanostructured materials as photocatalysts is often not accounted for in traditional wastewater treatment reactions. Particularly, some inorganic nanomaterials employed as photocatalysts may release secondary pollutants in the form of ionic species that leach out due to photocorrosion. In this context, this work is a proof-of-concept study for exploring the environmental toxicity effect of extremely small-sized nanoparticles (<10 nm) like quantum dots (QDs) that are employed as photocatalysts, and in this study, cadmium sulfide (CdS) QDs are chosen. Typically, CdS is an excellent semiconductor with suitable bandgap and band-edge positions that is attractive for applications in solar cells, photocatalysis, and bioimaging. However, the leaching of toxic cadmium (Cd2+) metal ions due to the poor photocorrosion stability of CdS is a matter of serious concern. Therefore, in this report, a cost-effective strategy is devised for biofunctionalizing the active surface of CdS QDs by employing tea leaf extract, which is expected to hinder photocorrosion and prevent the leaching of toxic Cd2+ ions. The coating of tea leaf moieties (chlorophyll and polyphenol) over the CdS QDs (referred to hereafter as G-CdS QDs) was confirmed through structural, morphological, and chemical analysis. Moreover, the enhanced visible-light absorption and emission intensity of G-CdS QDs in comparison to that of C-CdS QDs synthesized through a conventional chemical synthesis approach confirmed the presence of chlorophyll/polyphenol coating. Interestingly, the polyphenol/chlorophyll molecules formed a heterojunction with CdS QDs and enabled the G-CdS QDs to exhibit enhanced photocatalytic activity in the degradation of methylene blue dye molecules over C-CdS QDs while effectively preventing photocorrosion as confirmed from cyclic photodegradation studies. Furthermore, detailed toxicity studies were conducted by exposing zebrafish embryos to the as-synthesized CdS QDs for 72 h. Surprisingly, the survival rate of the zebrafish embryos exposed to G-CdS QDs was equal to that of the control, indicating a significant reduction in the leaching of Cd2+ ions from G-CdS QDs in comparison to C-CdS QDs. The chemical environment of C-CdS and G-CdS before and after the photocatalysis reaction was examined by X-ray photoelectron spectroscopy. These experimental findings prove that biocompatibility and toxicity could be controlled by simply adding tea leaf extract during the synthesis of nanostructured materials, and revisiting green synthesis techniques can be beneficial. Furthermore, repurposing the discarded tea leaves may not only facilitate the control of toxicity of inorganic nanostructured materials but can also help in enhancing global environmental sustainability.

Prophylactic and Therapeutic Role of Catechin-Loaded Poly(D,L-Lactic-Co-Glycolic Acid) Nanocapsules in Gastric Ulcer by In Vitro and In Vivo Approach

Background: Gastric ulcer develops from imbalance of gastro-aggressive and protective factors. As existing drugs have adverse effects, use of natural products is in continuous expansion. In this study, we prepared nanoformulation with catechin and polylactide-co-glycolide to provide a sustained, controlled and targeted delivery. Materials & methods: Detailed characterization and toxicity study of nanoparticles were done on cells and Wistar rats. The comparative actions of free compound and nanocapsule were investigated in vitro and in vivo during treatment of gastric injury. Results: Nanocatechin improved bioavailability, reduced gastric damage at a significantly lower dose (2.5mg/kg) by safeguarding from reactive oxygen species, restored mitochondrial integrityanddownregulated MMP-9 and other inflammatory mediators. Conclusion: Nanocatechin is a better alternative for preventing and healing gastric ulcers.

Геропротекторная активность транс-коричной кислоты, выделенной из шлемника байкальского (Scutellaria baicalensis)

Trans-cinnamic acid is a phenolic compound with a wide range of bioactive properties, including antioxidant and antibacterial effects. It also has high potential in the food and cosmetic industries. We aimed to isolate trans-cinnamic acid from the Baikal skullcap (Scutellaria baicalensis) and study its geroprotective activity on Caenorhabditis elegans nematodes used as a model organism.&#x0D; Our study objects included the S. baicalensis root culture and its extract, trans-cinnamic acid isolated from the extract, and C. elegans nematodes. Trans-cinnamic acid was isolated by high-performance liquid chromatography. The acid’s geroprotective activity was studied by evaluating its effect at concentrations of 10, 50, 100, and 200 μmol/L on the lifespan, stress resistance, and reproductivity of C. elegans. For the lifespan study, the nematodes were cultivated at 20°C for 61 days. To assess their resistance to oxidative stress, 15 μL of 1M paraquat was added to each well of the plate. Thermal stress resistance was determined by raising the temperature to 33°C. For the reproductivity study, the nematodes were cultivated in the S-medium with the addition of Escherichia coli OP50 and trans-cinnamic acid at required concentrations for 72 h.&#x0D; The maximum increase in lifespan (9.8%) was observed in the nematodes treated with 50 μmol/L of trans-cinnamic acid. Under oxidative stress, all the concentrations of trans-cinnamic acid increased the survival of nematodes, while under thermal stress, trans-cinnamic acid reduced the percentage of surviving nematodes. At a concentration of 100 μmol/L, trans-cinnamic acid increased the nematodes’ reproduction by 1.48 times.&#x0D; Based on our data, trans-cinnamic acid isolated from S. baicalensis can be recommended as a bioactive compound with geroprotective activity. However, further research is needed on other model organisms with detailed toxicity studies.

Bioactivity profiling of per- and polyfluoroalkyl substances (PFAS) identifies potential toxicity pathways related to molecular structure

Per- and polyfluoroalkyl substances (PFAS) are a broad class of hundreds of fluorinated chemicals with environmental health concerns due to their widespread presence and persistence in the environment. Several of these chemicals have been comprehensively studied for experimental toxicity, environmental fate and exposure, and human epidemiology; however, most chemicals have limited or no data available. To inform methods for prioritizing these data-poor chemicals for detailed toxicity studies, we evaluated 142 PFAS using an in vitro screening platform consisting of two multiplexed transactivation assays encompassing 81 diverse transcription factor activities and tested in concentration-response format ranging from 137 nM to 300 μM. Results showed activity for various nuclear receptors, including three known PFAS targets--specifically estrogen receptor alpha and peroxisome proliferator receptors alpha and gamma. We also report activity against the retinoid X receptor beta, the key heterodimeric partner of type II, non-steroidal nuclear receptors. Additional activities were found against the pregnane X receptor, nuclear receptor related-1 protein, and nuclear factor erythroid 2-related factor 2, a sensor of oxidative stress. Using orthogonal assay approaches, we confirmed activity of representative PFAS against several of these targets. Finally, we identified key PFAS structural features associated with nuclear receptor activity that can inform future predictive models for use in prioritizing chemicals for risk assessment and in the design of new structures devoid of biological activity.

Seasonal variations in atrazine degradation in a typical semienclosed bay of the northwest Pacific ocean

Pesticides are widely used to alleviate pest pressure in agricultural systems, and atrazine is a typical diffuse pollutant and serves a sensitivity index for environmental characteristics. Based on the physicochemical properties of parent substances, degradation products of pesticides may pose a greater threat to aquatic ecosystems than pesticides. Atrazine and three primary degradation products (deethylatrazine (DEA), deisopropylatrazine (DIA) and didealkylatrazine (DDA)) were investigated in a semienclosed bay of the western Pacific Ocean. Seasonal surface water and suspended particulate sediment (SPS) samples were collected from the estuary and bay in January, April, and August 2019. The level of pesticide contamination was lower in the bay than in the estuary, and the pesticide concentration in the dissolved phase was higher than that in the adsorbed phase. The average concentrations of atrazine and the three degradation products in the three seasons ranged from 2.42 to 328.46 ng/L in water and from 0.07 to 12.75 ng/L in SPS. The proportion of atrazine among the four detected pollutants decreased from 0.7 to 0.1 in surface water and from 0.3 to 0.1 in SPS over the seasons. As the main degradation products, the concentration proportions of DDA and DEA reached as high as 0.6 in August. The ratio of DEA to atrazine (DEA/ATR) increased from January to August, which indicated the progressive degradation process in the bay. Single-factor analysis of variance and principal component analysis indicated that atrazine degradation was sensitive to temperature, dissolved oxygen, and salinity. These three factors accounted for almost 70% of the seasonal variance in atrazine without a quantification assessment of photolysis or bacteria. The spatial distributions of DEA in the three seasons demonstrated that wind and currents also played important roles in pollutant redistribution. The seasonal temporal and spatial correlations between water and SPS demonstrated the degradation patterns of atrazine in marine conditions, supporting the need for future detailed toxicity studies.

Folic acid-appended galactoxyloglucan-capped iron oxide nanoparticles as a biocompatible nanotheranostic agent for tumor-targeted delivery of doxorubicin

Iron oxide nanoparticles (IONPs) are employed as MRI contrast agents and as effective drug delivery vehicles. However, the limited solubility and biodegradability of these nanoparticles need to be improved for safer biomedical applications. In an attempt to improve the bottlenecks associated with IONPs, the current study focuses on the synthesis of folic acid conjugated, galactoxyloglucan-iron oxide nanoparticles (FAPIONPs), for the loading and controlled release of the encapsulated chemotherapeutic agent doxorubicin (DOX). The as-designed DOX@FAPIONPs induced a dose-dependent increase in cytotoxicity in folate receptor-positive cells through a caspase-mediated programmed cell death pathway while bare DOX demonstrated a non-targeted toxicity profile. Using LC-MS/MS analysis, several major biological processes altered in treated cells, from which, cell cycle, cellular function and maintenance were the most affected. Detailed toxicity studies in healthy mice indicated the absence of any major side effects while bare drugs created substantial organ pathology. Gadolinium-based contrast agents have a risk of adverse effects, including nephrogenic systemic fibrosis overcome by the administration of DOX@FAPIONPs in xenograft mice model. Tumor-targeted biodistribution pattern with a favorable DOX pharmacokinetics will be the driving factor behind the appealing tumor reduction capacity and increased survival benefits demonstrated on solid tumor-bearing mice.

Genotoxic potential of a novel PDE-4B inhibitor Apremilast by chromosomal aberration and micronucleus assay in mice

ObjectiveResearchers have confirmed that chronic administration of drugs at high doses causes genotoxicity which serve as first step in development of cancers. Apremilast, a phosphodiesterase-4 inhibitor is Food and Drug Administration (FDA) approved drug for Psoriatic Arthritis. The present study designed to conduct genotoxicity testing using the genotoxic study which give simple, sensitive, economical and fast tools for the assessment of damage of genetic material. MethodsTo conduct genotoxicity study of Apremilast, 60 Swiss albino male mice divided into 6 groups (n = 10). Group1 served as a normal control group without any treatment, Group 2 treated as a disease control and administered with cyclophosphamide 40 mg/kg, IP. Group 3, 4, 5 and 6 treated as test groups and received 10, 20, 40 and 80 mg/kg/day Apremilast respectively. The total duration of study was 13 weeks. At termination day animals were sacrificed and chromosomal aberration assay (BMCAA) and micronucleus assay (BMMNA) were performed to know the genotoxicity potential of Apremilast. ResultsThe results indicates significant rise in chromosomal aberrations (CA) frequency in bone marrow cells and decrease in the MI of the disease control animals as well as Apremilast treated groups. Further significant (p < 0.001; p < 0.0001) increase in score of micronucleated polychromatic erythrocytes (MNPCEs) and percentage of micronucleated PCEs per 1000 PCEs and decrease in the ratio of polychromatic/normochromatic erythrocytes (PCE/NCE) was observed in micronucleus assay. Genotoxic effect increases with the increase of Apremilast dose. Conclusion: Finding of present indicates that Apremilast shows genotoxic potential on high administration although further detailed toxicity studies required for confirmations.

Using of the Synthetic Dural Graft as Described in Bogota Bag Technique to Decrease the Adhesions in Decompressive Craniectomy.

To create an alternative synthetic dural graft using a parenteral solution bag made of polyvinyl chloride/polypropylene (PVC/ PP). Twenty-two albino rats were divided into the Sham, DC and BAG groups. Except for the Sham group animals, the right parietal bone of the rats was totally drilled in a diameter of 1.0 x 0.5 mm. Then, the PVC/PP BAG was layered over the craniectomized bone of the BAG group animals. Thirty days later, all animals were sacrificed, and inflammatory processes consisting of polymorphonuclear cell infiltration, inflammation, edema, hyperemia, lymphocytosis, histiocytosis, vascular proliferation, and fibrosis were graded at the craniectomy site. The grade values of inflammation, edema, histiocytosis, and fibrosis were found different among the groups (p < 0.017). It was observed that placing a synthetic graft to the surgical site undergoing craniectomy could prevent fibrotic adhesions that might occur between the brain tissue and scalp in the chronic period. Furthermore, it was considered that this synthetic material did not increase inflammatory processes secondary to surgery at the surgical site and did not produce a foreign body reaction, toxicity, or infection. As a result of this study, it was argued that the synthetic material used in this study could be compatible with dermal and neural tissues and reduce adhesions at the craniectomy field. Therefore, it was considered that this material could be used as an alternative synthetic dural graft in decompressive craniectomy in human subjects after detailed toxicity studies.

Therapeutic potential of xanthones from Swertia chirata in breast cancer cells.

Background & objectives:Medicinal plants like Swertia chirata are rich sources of different xanthones. This study was aimed to assess the cytotoxic potential of four most abundant xanthones present in S. chirata both in vivo and in vitro in Ehrlich ascites carcinoma (EAC), a mouse transplantable breast carcinoma cell line and two human breast carcinoma cell lines (MCF-7 and MDA-MB-231).Methods:Four xanthones derived from S. chirata namely 1-hydroxy-3,7,8-trimethoxyxanthone (XA), 1,8-dihydroxy-3,5-dimethoxyxanthone (XB), 1-hydroxy-3,5,8-trimethoxyxanthone (XC) and 1,5,8-trihydroxy-3-methoxyxanthone (XD) were used for determination of sub-lethal dose on the cell lines EAC, MCF-7, MDA-MB-231 and verified toxicity of sub-lethal dose on normal murine fibroblast cells. Cytotoxicity was measured in vivo and survivability of mice was plotted accordingly. Therapeutic efficacy of XD was evaluated both in vivo and in vitro by determination of lipid peroxidation (LPO), reactive oxygen species (ROS) generation and by quantitating the enzyme status (GSH, catalase, superoxide dismutase) in treated and untreated samples. DNA damage was evaluated using comet and DNA fragmentation assays. Furthermore, apoptotic effect was analyzed by flow cytometry and validated by TUNEL assay and Western blotting.Results:Among all the xanthones tested XD showed IC50 at the lowest dose, and normal cells were unaffected at this dose. Survivability of mice increased significantly when treated with XD compared to other xanthones and cisplatin. Significantly increased ROS and LPO were found in cancer cells as a result of XD treatment which was unaltered in normal cell line. XD induced DNA damage and apoptosis in cancer cell lines.Interpretation & conclusions:Our experimental data indicate that XD may potentially act as a chemotherapeutic agent by enhancing ROS in breast cancer cells thereby leading to apoptosis.

Repeated dose (90 days) oral toxicity study of ursolic acid in Han-Wistar rats

BackgroundUrsolic acid (UA) has been used in alternative medicine for decades, and there has been a great interest in its medicinal properties. Despite this increased interest, a detailed long-term toxicity study has not been performed. The objective of this study was to determine the long-term toxic effect of UA on clinical chemistry, haematology, coagulation, pathology/morphology, behaviour and motor skills in rats. MethodsA solution was made by dissolving UA in a mixture of 0.1% Tween 80 and 0.5% hydroxypropyl methylcellulose in Milli-Q Water. The control group received the vehicle, and the test groups received a dose up to 1000 mg/kg/day via oral gavage. The solution was administered to both male and female (Han-Wistar) rats for 90 consecutive days. ResultsUA did not cause any deaths, abnormal body weights or abnormal pathology at all test doses. In addition to that, no toxicological changes were observed in behaviour, neurotoxicity, coagulation, haematology or clinical chemistry that are related to the administration of UA. ConclusionThis study indicates that oral dosing of UA for 90 consecutive days does not lead to toxic effects at any of the doses. Therefore, the NOAEL for UA is likely to be higher than 1000 mg/kg/day.

Battling Glioblastoma: A Novel Tyrosine Kinase Inhibitor with Multi-Dimensional Anti-Tumor Effect (Running Title: Cancer Cells Death Signalling Activation).

Glioblastoma (GB), a grade IV glioma, with high heterogeneity and chemoresistance, obligates a multidimensional antagonist to debilitate its competence. Considering the previous reports on thioesters as antitumor compounds, this paper investigates on use of this densely functionalized sulphur rich molecule as a potent anti-GB agent. Bio-evaluation of 12 novel compounds, containing α-thioether ketone and orthothioester functionalities, identified that five analogs exhibited better cytotoxic profile compared to standard drug cisplatin. Detailed toxicity studies of top compound were evaluated in two cell lines, using cell viability test, apoptotic activity, oxidative stress and caspase activation and RNA-sequencing analysis, to obtain a comprehensive molecular profile of drug activity. The most effective molecule presented half maximal inhibitory concentration (IC50) values of 27 μM and 23 μM against U87 and LN229 GB cells, respectively. Same compound effectively weakened various angiogenic pathways, mainly MAPK and JAK-STAT pathways, downregulating VEGF. Transcriptome analysis identified significant promotion of apoptotic genes, and genes involved in cell cycle arrest, with concurrent inhibition of various tyrosine kinase cascades and stress response genes. Docking and immunoblotting studies suggest EGFR as a strong target of the orthothioester identified. Therefore, orthothioesters can potentially serve as a multi-dimensional chemotherapeutic possessing strong cytotoxic, anti-angiogenic and chemo-sensitization activity, challenging glioblastoma pathogenesis.

Acute oral toxicity of the ellagitannin geraniin and a geraniin-enriched extract from Nephelium lappaceum L rind in Sprague Dawley rats

Despite the lack of its toxicity evaluation, traditional herbal products are being widely used for various health indications. Geraniin, an ellagitannin, is a bioactive compound found in many traditional herbal medicines. In spite its numerous health benefits ranging from anti-inflammatory, anti-hyperglycaemic, hepatoprotective, anti-cancer and anti-microbial, no toxicity data on geraniin is available. The objective of this study is to evaluate the acute oral toxicity of geraniin and an enriched geraniin-extract of Nephelium lappaceum L rind. This study followed the guidelines of the OECD 423 acute oral toxicity test. Subsequent to a single oral administration of the test compounds, the rats were observed for 14 days for signs of toxicity and mortality. Following euthanasia, full blood count, biochemistry of blood and histopathology assessment of organs were carried out. All parameters analysed indicated insignificant difference compared to control. The LD50 cut-off values for both geraniin and geraniin-enriched extract was established to be 2000 mg/kg b. w., following a single oral dose. It was however observed that the hepatocytes of three geraniin-administered rats exhibited a ‘foamy appearance’. As such, the no-observed-adverse-effect level of geraniin is below 2000 mg/kg, while that of geraniin-enriched extract is up to 2000 mg/kg. Further detailed toxicity studies are required to establish geraniin or its enriched extract from Nephelium lappaceum L rind safe for human consumption.

Heavy metal content and health risk assessment of commonly patronized herbal medicinal preparations from the Kumasi metropolis of Ghana.

To address the question of whether users of herbal products (HPs) are exposed to harmful contaminants, we evaluated six HPs mostly patronized in Kumasi for heavy metal contamination and assessed the health risk associated with their use. This study is one of the first safety evaluation studies on finished multiherbal products in the region. Three antimalarial, two antidiabetic and one antihypertensive HPs were selected after a mini-survey and coded randomly as HP A-F. The HPs were acid digested for quantitative analysis of heavy metals using Inductively Coupled Plasma Mass Spectrometer. Hg quantification was carried out using cold vapour atomic absorption spectroscopy. The cancer risk estimation values for the carcinogenic metals ranged between 1.54 × 10-9 to 3.73 × 10-4 and were all within acceptable limits. The non-cancer health risk evaluation revealed that, some of the products pose health risk to consumers. The estimated daily intake (EDI) for As in HPF was 2.48 × 10-4 mg/kg/day compared to the reference limit of 1.67 × 10-4 mg/kg/day. HPF also had high hazard index (HI) of 5.70 (HI >1) in children as compared to 1.68 (HI >1) in adults showing a 3.4 folds increase in the health risk among the former. The six polyherbal products exhibited carcinogenic risk within acceptable limits. Although, the non-carcinogenic risk assessment of products HPA to HPE suggests safety, this can only be ascertained after further characterization of their health risks in detailed chronic toxicity studies. The high HI for product HPF suggests health risk for consumers of this product.

Toxicity bioassay of waste cooking oil-based biodiesel on marine microalgae.

The world biodiesel production is increasing at a rapid rate. Despite its perceived safety for the environment, more detailed toxicity studies are mandatory, especially in the field of aquatic toxicology. While considerable attention has been paid to biodiesel combustion emissions, the toxicity of biodiesel in the aquatic environment has been poorly understood. In our study, we used an algae culture growth-inhibition test (OECD 201) for the comparison of the toxicity of B100 (pure biodiesel), produced by methanol transesterification of waste cooking oil (yellow grease), B0 (petroleum diesel fuel) and B20 (diesel-biodiesel blended of 20% biodiesel and 80% petroleum diesel fuel by volume). Two marine diatoms Attheya ussuriensis and Chaetoceros muelleri, the red algae Porphyridium purpureum and Raphidophyte Heterosigma akashiwo were employed as the aquatic test organisms. A sample of biodiesel from waste cooking oil without dilution with petroleum diesel (B100) showed the highest level of toxicity for the microalgae A. ussuriensis, C. muelleri and H. akashiwo, compared to hexane, methanol, petroleum diesel (B0) and diluted sample (B20). The acute EC50 in the growth-inhibition test (96 h exposure) of B100 for the four species was in the range of 3.75-23.95 g/L whereas the chronic toxicity EC50 (7d exposure) was in the range of 0.42-16.09 g/L.

A comprehensive LC–MS/MS method validation for the quantitative investigation of 37 fingerprint phytochemicals in Achillea species: A detailed examination of A. coarctata and A. monocephala

The current study aims to optimize and validate a comprehensive LC–MS/MS method for the quantification of 37 phytochemicals (15 phenolic acids, 17 flavonoids, 3 non-phenolic organic acids, 1 phenolic aldehyde and 1 benzopyrene) in Achillea species. Though Achillea species were chosen as real life samples, the current method is applicable to a wide range of plant species. The developed method was fully validated in terms of linearity, accuracy (recovery), inter-day and intra-day precision (repeatability), limits of detection and quantification (LOD/LOQ) and relative standard uncertainty (U% at 95% confidence level (k = 2)). Reversed-phase ultrahigh performance liquid chromatography was optimized to achive optimum separation for 37 phytochemical compounds and to overcome the suppression effects. MS detection was performed using a triple quadrupole mass spectrometer and negative or positive ionization modes were optimized for each analyte. Multiple reaction monitoring (MRM) was used to quantify the analytes, related molecular ions and transition ions were optimized. Phytochemical screening of ethanol and methanol-chloroform extracts of root and aerial parts of A. coarctata and A. monocephala were performed by using the developed and validated LC–MS/MS method. Root and aerial parts of both species have considerable amounts of certain phenolic-nonphenolic acids (quinic, malic, fumaric, chlorogenic and vanillic acids) and flavonoids (rutin, hesperidin, isoquercitrin, apigetrin, luteolin, apigenin). Additionally, total phenolic and flavonoid amounts, antioxidant (DPPH free radical scavenging assay, ABTS radical cation decolorization assay, β-carotene lipid peroxidation test system and CUPRAC cupper reduction capacity methods), anticholinesterase, tyrosinase, urease inhibition and cytotoxic activities (on HeLa (Human Cervical Carcinoma Cell Line) of A. coarctata and A. monocephala were also investigated. It has been determined that the studied Achillea species, that are rich in total phenolic-flavonoid and chlorogenic acid contents, have high antioxidant and cytotoxic potential at the same time. According to the results of LC–MS/MS, antioxidant and cytotoxic activity studies, after detailed chemical investigation and toxicity studies on these species, A. coarctata and A. monocephala may be promoted as promising sources of natural agents and used for the development of nutraceuticals or functional food ingredients in future.

Copper Manganese Sulfide Nanoplates: A New Two-Dimensional Theranostic Nanoplatform for MRI/MSOT Dual-Modal Imaging-Guided Photothermal Therapy in the Second Near-Infrared Window.

Multifunctional nanoplatforms with integrated diagnostic and therapeutic functions have attracted tremendous attention. Especially, the second near-infrared (NIR-II) light response-based nanoplatforms hold great potential in cancer theranostic applications, which is because the NIR-II window provides larger tissue penetration depth and higher maximum permissible exposure (MPE) than that of the well-studied first near-infrared (NIR-I) window. Herein, we for the first time present a two-dimensional (2D)-nanoplatform based on Cu2MnS2 nanoplates (NPs) for magnetic resonance imaging (MRI)/multispectral optoacoustic tomography (MSOT) dual-modal imaging-guided photothermal therapy (PTT) of cancer in the NIR-II window. Methods: Cu2MnS2 NPs were synthesized through a facile and environmentally friendly process. A series of experiments, including the characterization of Cu2MnS2 NPs, the long-term toxicity of Cu2MnS2 NPs in BALB/c nude mice, the applications of Cu2MnS2 NPs for in vitro and in vivo MRI/MSOT dual-modal imaging and NIR-II PTT of cancer were carried out. Results: The as-synthesized Cu2MnS2 NPs exhibit low cytotoxicity, excellent biocompatibility as well as high photothermal conversion efficiency (~49.38%) and outstanding photostability. Together with their good T1-shortening effect and strong absorbance in the NIR-I and NIR-II region, the Cu2MnS2 NPs display high-contrast imaging performance both in MRI and MSOT (900 nm laser source). Moreover, the subsequent in vitro and in vivo results demonstrate that the Cu2MnS2 NPs possess excellent PTT efficacy under 1064 nm laser irradiation with a low power density (0.6 W cm-2). In addition, the detailed long-term toxicity studies further confirming the safety of Cu2MnS2 NPs in vivo. Conclusion: We have developed a new 2D Cu2MnS2 NPs as multifunctional theranostic agents for MRI/MSOT dual-modal imaging-guided PTT of cancer in the NIR-II window. Such biocompatible Cu2MnS2 NPs might provide a new perspective for exploring new 2D-based nanoplatforms with improved properties for clinical applications in the future.

Antiamoebic Activity of Adenophyllum aurantium (L.) Strother and Its Effect on the Actin Cytoskeleton of Entamoeba histolytica.

In Mexico, the Adenophyllum aurantium (L.) Strother plant is consumed as an infusion to treat intestinal diseases such as amoebiasis, which is an endemic health problem in Mexico and other countries. However, the effect of A. aurantium on Entamoeba histolytica, the causative agent of amoebiasis, is unknown. An aerial part methanolic extract (AaMeA), a root methanolic extract (AaMeR) and a root ethyl acetate extract (AaEaR) were tested on E. histolytica trophozoites. AaMeA and AaMeR did not show antiproliferative activity; however, AaEaR exhibited an in vitro GI50 of 230 μg/ml, and it was able to inhibit the differentiation of Entamoeba invadens trophozoites into cysts. The intraperitoneal administration of AaEaR (2.5 or 5 mg) to hamsters that were infected with E. histolytica inhibited the development of amoebic liver abscesses in 48.5 or 89.0% of the animals, respectively. Adhesion to fibronectin and erythrophagocytosis were 28.7 and 37.5% inhibited by AaEaR, respectively. An ultrastructure analysis of AaEaR-treated trophozoites shows a decrease in the number of vacuoles but no apparent cell damage. Moreover, this extract affected the actin cytoskeleton structuration, and it prevented the formation of contractile rings by mechanism(s) that were independent of reactive oxygen species and RhoA activation pathways. 13C NMR data showed that the major compounds in the AaEaR extract are thiophenes. Our results suggest that AaEaR may be effective in treatments against amoebiasis, nevertheless, detailed toxicity studies on thiophenes, contained in AaEaR, are required to avoid misuse of this vegetal species.

Abstract A22: NEO212: A novel temozolomide-perillyl alcohol conjugate exhibits superior activity against temozolomide resistant gliomas and intracranial triple negative breast cancer metastasis

Abstract Although the alkylating agent temozolomide (TMZ) has become the standard of care in the treatment of malignant gliomas, its overall efficacy is still limited by development of drug resistance, limited blood brain barrier (BBB) penetrance, and myelotoxicity. Recently, we have synthesized a TMZ analog where the monoterpene perillyl alcohol (POH) is covalently linked to TMZ via a carbamide bond. This new compound (NEO212) has been tested in two situations: TMZ resistant gliomas and breast cancer intracranial metastasis. Glioma cell lines (U251TR, LN229TR2, T98G) determined to be TMZ resistant based on base excision repair (BER) pathway, mismatch repair (MMR) deficiency, or overexpression of O6 methyl-guanine-DNA methyltransferase (MGMT) were evaluated in-vitro. In vivo experiments were performed using intracranial implant of luciferase labeled TMZ resistant cell lines. Triple negative breast cancer lines were tested, and implanted intracranially for in-vivo model. NEO212 was administered in both cases using subcutaneous injection using 10 day treatment, 7 day rest cycles did not have significant toxicity on normal astrocytes and brain endothelial cells. NEO212 induced cell death appeared to be independent of the mechanism of DNA repair resistance. In vivo experiments using intracranial implant of luciferase labeled TMZ resistant cell lines demonstrated that NEO212 crossed the BBB, reduced intracranial growth, and increased animal survival significantly compared to TMZ, without significant toxicity (including bone marrow). NEO212 was also effective in TMZ resistant breast cancer lines. In an intracranial mouse tumor model with triple-negative breast cancer, NEO212 revealed considerably greater therapeutic efficacy than TMZ, where a single cycle of treatment (10 days) extended median survival benefit from 6 days (in the case of TMZ) to 28 days, with good tolerance. NEO212 appears to be a promising well tolerated new agent with similar mechanism of cytotoxicity to TMZ. Its increased potency is most likely multi-factorial: 1) increased DNA damage, involving a broader scope of DNA repair mechanisms. 2) linkage with POH, resulting in longer biological half-life and greater opportunity for placement of cytotoxic DNA lesions. 3) increased lipophilicity, allowing for better penetration of the BBB, and possibly cell membrane. Preliminary results demonstrate that it can safely be administered systemically; more detailed toxicity studies are currently underway. Citation Format: Thomas C. Chen, Axel H. Schonthal, Weijun Wang, Heeyon Cho, Florence H. Hofman. NEO212: A novel temozolomide-perillyl alcohol conjugate exhibits superior activity against temozolomide resistant gliomas and intracranial triple negative breast cancer metastasis. [abstract]. In: Proceedings of the AACR Special Conference: Advances in Brain Cancer Research; May 27-30, 2015; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2015;75(23 Suppl):Abstract nr A22.